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Topics in Current Genetics Series Editor: Stefan Hohmann
2
Springer Berlin Heidelberg New York Hong Kong London Milan Paris Tokyo
Johannes H. de Winde
(Editor)
Functional Genetics of Industrial Yeasts
With 50 Figures
123
Professor JOHANNEs H. dE WINDE Beyerinck Laboratory DSM Life Sciences Bakery Ingredients Technology PO Box 1 2600 MA Delft The Netherlands and Kluyver Laboratory for Biotechnology Technical University of Delft Julianalaan 67 2628 BC Delft The Netherlands
ISSN 1610-2096 ISBN 3-540-02489-1 Springer-Verlag Berlin Heidelberg New York Cataloging-in-Publication Data applied for Bibliographic information published by Die Deutsche Bibliothek Die Deutsche Bibliothek lists this publication in the Deutsche Nationalbibliografie; detailed bibliographic data is available in the Internet at This work is subject to copyright. All rights reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilm or in any other way, and storage in data banks. Duplication of this publication or parts thereof is permitted only under the provisions of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer-Verlag. Violations are liable for prosecution under the German Copyright Law. Springer-Verlag Berlin Heidelberg New York a member of BertelsmannSpringer Science+Business Media GmbH http://www.springer.de Springer-Verlag Berlin Heidelberg 2003 Printed in Germany The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Typesetting: Camera ready by editors Data-conversion: PTP-Berlin, Stefan Sossna e.K. Cover Design: Design & Production, Heidelberg 39/3150-WI - 5 4 3 2 1 0 - Printed on acid-free paper
Table of contents
1 Functional genetics of industrial yeasts; of ancient skills and modern applications .........................................................................................................1 Han de Winde ....................................................................................................1 Abstract .........................................................................................................1 1.1 Introduction .............................................................................................1 1.1.1 History .............................................................................................1 1.1.2 Yeast diversity .................................................................................2 1.1.3 Yeast biotechnology and yeast genetics...........................................3 1.1.4 Set-up of this book...........................................................................4 1.2 From classical genetics to modern genomics ..........................................5 1.3 Improving ancient skills: Saccharomyces cerevisiae in food and beverage ........................................................................................................6 1.3.1 Baker’s yeast....................................................................................6 1.3.2 Wine yeast .......................................................................................7 1.3.3 Brewer’s yeast..................................................................................8 1.4 Of apples and pears: new food yeast .......................................................8 1.4.1 Kluyveromyces lactis .......................................................................9 1.4.2 Torulaspora delbrueckii...................................................................9 1.5 Yeast as model system ..........................................................................10 1.5.1 Saccharomyces yeast in signal transduction and cell wall biosynthesis.............................................................................................10 1.5.2 Kluyveromyces lactis .....................................................................10 1.5.3 Hansenula, Pichia, and Yarrowia yeasts .......................................11 1.6 Novel biotechnological tools and applications ......................................11 1.6.1 Yeast in biocontrol.........................................................................11 1.6.2 Metabolic pathway engineering .....................................................11 1.7 Perspectives, rules and regulations........................................................13 Acknowledgements .....................................................................................13 References ...................................................................................................14 2 Genetics and classical genetic manipulations of industrial yeasts................17 Paul V. Attfield and Philip J. L. Bell ...............................................................17 Abstract .......................................................................................................17 2.1 Introduction ...........................................................................................17 2.2 Genetic properties of industrial yeasts...................................................19 2.2.1 An overview of the genome and functional genetic analysis of laboratory S. cerevisiae strains: a basis for comparison with industrial yeasts ......................................................................................................19 2.2.2 Genomes of industrial Saccharomyces and non-Saccharomyces yeasts ......................................................................................................24 2.2.3 Functional analyses of industrially relevant yeasts ........................26
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2.2.4 Classical genetic features of laboratory and industrial yeasts........ 28 2.3 Industrial yeast strain improvement ...................................................... 30 2.3.1 Issues affecting the choice of genetic strategy............................... 30 2.3.2 Mating............................................................................................ 31 2.3.3 Mutagenesis ................................................................................... 35 2.3.4 Protoplast / spheroplast fusion....................................................... 37 2.3.5 Cytoduction ................................................................................... 38 2.3.6 Screening strains for improvements............................................... 38 2.4 Improvements to strains via classical genetics ...................................... 39 2.4.1 Improved maltose utilization in industrial S. cerevisiae: a union of functional analysis and classical genetics in strain improvement ........... 40 2.5 Conclusions and prospects .................................................................... 46 References................................................................................................... 46 3 Baker’s yeast: challenges and future prospects ............................................. 57 Francisca Randez-Gil, Jaime Aguilera, Antonio Codón, Ana M. Rincón, Francisco Estruch and Jose A. Prieto............................................................... 57 Abstract ....................................................................................................... 57 3.1 Introduction ........................................................................................... 57 3.2 Genetic characteristics of baker’s yeast strains ..................................... 59 3.2.1 Nuclear genome ............................................................................. 59 3.2.2 Sporulation..................................................................................... 62 3.3 Important traits for baking applications ................................................ 62 3.3.1 Fermentative characteristics .......................................................... 62 3.3.2 Osmotolerance and Na+ toxicity resistance.................................... 67 3.3.3 Cryoresistance ............................................................................... 75 3.4 non-Saccharomyces baker’s yeast: Torulaspora delbrueckii ................ 83 3.4.1 Morphological and genetic characteristics..................................... 83 3.4.2 Baking applications ....................................................................... 83 3.4.3 T. delbrueckii as a model system ................................................... 84 3.5 Conclusions ........................................................................................... 85 Acknowledgements ..................................................................................... 85 References................................................................................................... 85 4 The genetic analysis and tailoring of wine yeasts .......................................... 99 Isak S. Pretorius............................................................................................... 99 Abstract ....................................................................................................... 99 4.1 Introduction ........................................................................................... 99 4.2 The need for improved wine yeast strains........................................... 100 4.2.1 The advantages and disadvantages of spontaneous and inoculated fermentations ........................................................................................ 100 4.2.2 The development of active dried wine yeast starter culture strains101 4.3 The genetic features, analysis and modifications of wine yeasts ........ 102 4.3.1 The morphology, reproduction and genetic constitution of wine yeasts .................................................................................................... 102
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4.3.2 The genetic methods for the analysis and modification of wine yeasts ....................................................................................................104 4.4 Strategies and targets for the improvement of wine yeasts .................109 4.4.1 Improvement of fermentation performance .................................111 4.4.2 Improvement of processing efficiency.........................................116 4.4.3 Improvement of wine wholesomeness .........................................120 4.4.4 Improvement of wine flavour and other sensory qualities ...........125 4.5 Conclusions and future perspectives ...................................................130 Acknowledgements ...................................................................................133 References .................................................................................................134 5 Brewer’s yeast: genetic structure and targets for improvement ................143 Jørgen Hansen and Morten C. Kielland-Brandt.............................................143 Abstract .....................................................................................................143 5.1 The role of yeast in beer production....................................................143 5.2 Brewer’s yeast: a chimera in service ...................................................145 5.3 How to breed brewer’s yeast ...............................................................149 5.4 Targets and approaches in breeding of brewer’s yeast ........................151 5.4.1. Feeding the beast: carbohydrate fermentation ............................151 5.4.2 Flavour components: too little and too much...............................153 5.4.3. Flavour stability: a way to increased shelf life............................156 5.4.4 Diacetyl and maturation: how to speed things up ........................157 5.4.5 Sedimentation and filtration.........................................................159 5.5 Brewer’s yeast in the post-genomic era...............................................160 5.6 Concluding remarks ............................................................................160 Acknowledgements ...................................................................................162 References .................................................................................................162 6 Kluyveromyces lactis: genetics, physiology, and application .......................171 Karin D. Breunig and H. Yde Steensma ........................................................171 Abstract .....................................................................................................171 6.1 Introduction .........................................................................................171 6.2 Genetics...............................................................................................172 6.2.1 Taxonomy of Kluyveromyces ssp. and phylogenetic relationship to S. cerevisiae ..........................................................................................172 6.2.2 Chromosomes and extrachromosomal genetic elements..............173 6.2.3 Mitochondrial DNA .....................................................................176 6.2.4 Mating types ................................................................................176 6.3 Physiology...........................................................................................177 6.3.1 Carbon and energy metabolism....................................................177 6.3.2 Lactose utilization........................................................................188 6.3.3 The petite-negative phenotype .....................................................190 6.3.4 Killer Strains ................................................................................191 6.4. Industrial applications ........................................................................192 References .................................................................................................193
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7 The methylotrophic yeasts Hansenula polymorpha and Pichia pastoris: favourable cell factories in various applications.......................................... 207 Meis van der Heide, Marten Veenhuis and Ida van der Klei ......................... 207 Abstract ..................................................................................................... 207 7.1 Introduction ......................................................................................... 207 7.2 Tools for the production of heterologous proteins in Hansenula polymorpha and Pichia pastoris ............................................................... 209 7.2.1 Transformation and vectors ......................................................... 209 7.2.2 Expression cloning strategies....................................................... 209 7.2.3 Promoter systems for protein production..................................... 210 7.3 Sorting of heterologous proteins to specific subcellular locations ...... 212 7.3.1 Protein sorting to peroxisomes .................................................... 213 7.3.2 Protein secretion .......................................................................... 218 7.4 Concluding remarks ............................................................................ 219 Acknowledgements ................................................................................... 219 References................................................................................................. 219 8 Functional genetics of Yarrowia lipolytica .................................................... 227 Gerold Barth, Jean-Marie Beckerich, Angel Dominguez, Stefan Kerscher, David Ogrydziak, Vladimir Titorenko and Claude Gaillardin....................... 227 Abstract ..................................................................................................... 227 8.1 Introduction ......................................................................................... 227 8.2 Genetics and genetic tools................................................................... 228 8.2.1.Genetics ....................................................................................... 228 8.2.2 Genetic engineering ..................................................................... 229 8.2.3 Genomics ..................................................................................... 231 8.3 Yarrowia lipolytica as a model for protein secretion .......................... 231 8.3.1 Predominance of a cotranslational translocation pathway from cytoplasm to ER.................................................................................... 232 8.3.2 Sls1p, prototype of a new ADP/ATP exchange factor family for eukaryotic Hsp70p................................................................................ 232 8.3.3 Other new components affecting secretion.................................. 234 8.3.4 Function of the S. cerevisiae homologues of SEC genes ............. 235 8.4 Yarrowia lipolytica as a morphogenetic model................................... 236 8.4.1 Morphogenesis in Yarrowia lipolytica......................................... 236 8.4.2 Experimental approaches for the molecular cloning of genes involved in the yeast-hypha transition in Yarrowia lipolytica .............. 236 8.4.3 Identification and characterization of Y. lipolytica morphogenetic genes ..................................................................................................... 236 8.4.4 Pathway conservation in Y. lipolytica, S. cerevisiae, and C. albicans239 8.5 Yarrowia lipolytica as a model for mitochondrial complex I study .... 240 8.5.1 Mitochondrial metabolism........................................................... 240 8.5.2 Mitochondrial respiratory chain................................................... 242 8.5.3 Mitochondrial genome................................................................. 243 8.5.4 Respiratory chain complex I ........................................................ 244 8.5.5 The hydrogenase model for the catalytic core of complex I ........ 246
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8.6 Carbon metabolism in Yarrowia lipolytica .........................................247 8.6.1 Utilisation of carbohydrates by Yarrowia lipolytica ....................247 8.6.2 Utilisation of alcohols ..................................................................248 8.6.3 Utilisation of monocarboxylic acids ............................................248 8.6.4 Accumulation of storage carbohydrates.......................................249 8.6.5 Utilization of hydrocarbons as carbon source..............................249 8.6.6 Hydrolysis of fats.........................................................................250 8.6.7 Fatty acids biosynthesis and degradation.....................................251 8.7 Peroxisome assembly in the yeast Yarrowia lipolytica .......................251 8.7.1 Metabolic functions and biogenesis of peroxisomes....................252 8.7.2 Peroxisome biogenesis as it is presented in cell-biology textbooks253 8.7.3. A revision of the peroxisome biogenesis paradigm: peroxisomes assemble by a multistep pathway..........................................................253 8.7.4 Peroxisome fusion........................................................................254 8.7.5 The endoplasmic reticulum plays an essential role in peroxisome assembly ...............................................................................................256 8.7.6 Folded, oligomeric proteins are imported into peroxisomes........257 8.8 Future prospects ..................................................................................258 Acknowledgements ...................................................................................258 References .................................................................................................259 9 Yeasts and food spoilage ................................................................................273 S. Brul, J. van der Vossen, A. Boorsma, and F.M. Klis.................................273 Abstract .....................................................................................................273 9.1 Introduction to microbial and yeast food spoilage ..............................273 9.2 The ecology of food spoilage yeasts....................................................275 9.2.1 Diversity of yeasts .......................................................................275 9.2.2 Growth characteristics .................................................................275 9.2.3 Growth physiology ......................................................................276 9.2.4 Preservation strategies .................................................................276 9.3 The yeast envelope, antifungal targets, and (preservation) stress resistance ...................................................................................................277 9.3.1 Cell wall sugars and their synthesis .............................................279 9.3.2 Membrane localised transporters of low molecular weight compounds............................................................................................280 9.3.3 Cell wall proteins and stress response..........................................282 9.3.4 Sensing the extracellular environment and signalling stress........284 9.4. A new method for the analysis of gene expression profiles ...............285 9.5 Concluding remarks and future prospects ...........................................287 Acknowledgements ...................................................................................288 References .................................................................................................288 10 Non-conventional yeasts in antifungal application ....................................297 Volkmar Passoth and Johan Schnürer............................................................297 Abstract .....................................................................................................297 10.1 Introduction .......................................................................................297
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10.2 Antifungal yeasts and their modes of inhibition................................ 299 10.2.1 Phyllosphere yeasts.................................................................... 299 10.2.2 Biocontrol of postharvest diseases............................................. 300 10.2.3 Non-conventional killer yeasts in biocontrol ............................. 305 10.3 Available information about genome organisation in non-conventional antifungal yeasts........................................................................................ 306 10.3.1 Sexuality, genome structure, and extrachromosomal elements . 306 10.3.2 Genetics of the killer character in non-conventional biocontrol yeasts .................................................................................................... 309 10.4 Available genetic methods for biocontrol yeasts............................... 310 10.4.1 Molecular methods for identification and phylogenetic analysis of biocontrol yeasts ................................................................................... 311 10.4.2 Methods for the manipulation of biocontrol yeasts.................... 314 10.5 The use of genetic methods for the manipulation of biocontrol activity316 10.5.1 Application of mutants in biocontrol systems............................ 316 10.5.2 Application of genetically engineered strains in biocontrol systems ................................................................................................. 317 10.6 Conclusions and outlook ................................................................... 318 Acknowledgements ................................................................................... 318 References................................................................................................. 319 11 Yeast functional genomics and metabolic engineering: past, present and future .................................................................................................................. 331 Christoffer Bro, Birgitte Regenberg, and Jens Nielsen.................................. 331 Abstract ..................................................................................................... 331 11.1 Introduction ....................................................................................... 331 11.2 Examples of metabolic engineering .................................................. 333 11.2.1 Extension of substrate range ...................................................... 335 11.2.2 Heterologous protein production ............................................... 335 11.2.3 Improvement of fluxes............................................................... 336 11.3 Challenges in metabolic engineering ................................................ 338 11.4 Functional genomics ......................................................................... 340 11.4.1 Comparative sequence analysis ................................................. 341 11.4.2 Transcriptome analysis .............................................................. 342 11.4.3 Global mutant analysis .............................................................. 343 11.4.4 Proteome analysis ...................................................................... 344 11.4.5 Interactome analysis .................................................................. 345 11.4.6 Metabolome analysis ................................................................. 346 11.4.7 Fluxome analysis ....................................................................... 346 11.5 Functional genomics in metabolic engineering ................................. 347 11.5.1 Need for integrated approach..................................................... 350 11.6 Future prospects ................................................................................ 351 Acknowledgements ................................................................................... 352 References................................................................................................. 352 Index ................................................................................................................... 361
List of contributors Aguilera, Jaime Departamento de Biotecnología, IATA, CSIC, PO Box 73, 46100 Burjassot, Valencia, Spain Attfield, Paul Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia and Microbiogen Pty Ltd., Sydney, NSW 2109, Australia Barth, Gerold Institut für Mikrobiologie, Technische Universität Dresden, Mommsenstrasse 13, D-01062 Dresden, Germany Bekkerich Jean-Marie Génétique Moléculaire et Cellulaire, Institut National Agronomique ParisGrignon, CNRS INRA, 78850 Thiverval Grignon, France Bell, Philip, J.L. Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia Boorsma, A. Molecular Biology & Microbial Food Safety, University of Amsterdam, Swammerdam Institute for Life Sciences Nieuwe Achtergracht 166, NL-1018 WV Amsterdam, The Netherlands Breunig, Karin Institut für Genetik, Martin Luther Universität Halle-Wittenberg, 06099 Halle, Germany Bro, Christoffer Center for Process Biotechnology, BioCentrum DTU, Technical University of Denmark, Building 223, DK-2800 Lyngby, Denmark Brul, Stanley Molecular Biology & Microbial Food Safety, University of Amsterdam, Swammerdam Institute for Life Sciences Nieuwe Achtergracht 166, NL-1018 WV Amsterdam, The Netherlands and Food Processing Group, Unilever Research, Olivier van Noortlaan 120, NL-3133 AT Vlaardingen, The Netherlands
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Codón, Antonio Departamento de Genetica, Facultad de Biología, Universidad de Sevilla, PO Box 1095, 41080 Sevilla, Spain de Winde, Johannes H. Kluyver Laboratory for Biotechnology, Delft Technical University, Julianalaan 67, NL-2628 BC Delft, The Netherlands and DSM Bakery Ingredients, Technology Cluster, PO Box 1, bag 624-0265, NL-2600 MA Delft, The Netherlands Dominguez, Angel Departamento de Microbiologia y Genetica, Universidad de Salamanca, Plaza de los Doctores de la Reina s/n, 37007 Salamanca, Spain Estruch, Francisco Departamento de Biotecnología, IATA, CSIC, PO Box 73, 46100 Burjassot, Valencia, Spain Gaillardin, Claude Génétique Moléculaire et Cellulaire, Institut National Agronomique ParisGrignon, CNRS INRA, 78850 Thiverval Grignon, France Hansen, Jørgen Poalis A/S, Bülowsvej 25, DK-1870 Frederiksberg C, Denmark Klis, Frans Molecular Biology & Microbial Food Safety, University of Amsterdam, Swammerdam Institute for Life Sciences Nieuwe Achtergracht 166, NL-1018 WV Amsterdam, The Netherlands Kerscher, Stefan Universitätsklinikum Frankfurt, Institut für Biochemie I, Zentrum der biologischen Chemie, D-60590 Frankfurt am Main, Germany Kielland-Brandt, Morten C. Department of Physiology, Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-2500 Copenhagen Valby, Denmark Nielsen, Jens Center for Process Biotechnology, BioCentrum DTU, Technical University of Denmark, Building 223, DK-2800 Lyngby, Denmark Ogrydziak, David Institute of Marine Resources, University of California, Davis, CA 95616, USA
List of contributors
XIII
Passoth, Volkmar Deptartment of Microbiology, Swedish University of Agricultural Sciences, Box 7025, S-750 07 Uppsala, Sweden Pretorius, Isac S. The Australian Wine Research Institute, P.O. Box 197, Glen Osmond, Adelaide, SA 5064, Australia Prieto, José A. Departamento de Biotecnología, IATA, CSIC, PO Box 73, 46100 Burjassot, Valencia, Spain Randez-Gil, Francesca Departamento de Biotecnología, IATA, CSIC, PO Box 73, 46100 Burjassot, Valencia, Spain Regenberg, Birgitte Center for Process Biotechnology, BioCentrum DTU, Technical University of Denmark, Building 223, DK-2800 Lyngby, Denmark Rincón, Ana M. Departamento de Genetica, Facultad de Biología, Universidad de Sevilla, PO Box 1095, 41080 Sevilla, Spain Schnürer, Johan Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, S-750 07 Uppsala, Sweden Steensma, H. Yde Institute of Biology, Leiden University, Wassenaarseweg 64, NL-2333 AL Leiden, The Netherlands Titorenko, Vladimir Biology Department, Concordia University, 1455 de Maisonneuve Boulevard, West Montreal, Quebec, Canada H3G 1M8 van der Heide, Meis Eukaryotic Microbiology, Biomolecular Sciences and Biotechnology Institute, University of Groningen, PO Box 14, NL-9750 AA Haren, The Netherlands van der Klei, Ida J. Eukaryotic Microbiology, Biomolecular Sciences and Biotechnology Institute, University of Groningen, PO Box 14, NL-9750 AA Haren, The Netherlands
XIV
van der Vossen, J Risk Management & Microbiology, TNO Food Research, P.O. Box 360, NL3700 AJ Zeist, The Netherlands Veenhuis, Marten Eukaryotic Microbiology, Biomolecular Sciences and Biotechnology Institute, University of Groningen, PO Box 14, NL-9750 AA Haren, The Netherlands
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1.1.3 YGCUV DKQVGEJPQNQI[ CPF [GCUV IGPGVKEU &WTKPI VJG NCUV VYQ FGECFGU, VJG GZRNQKVCVKQP CPF WVKNK\CVKQP QH [GCUVU HQT DKQVGEJPQNQIKECN RWTRQUGU JCU RTQITGUUKXGN[ QWVTGCEJGF DG[QPF VJG DQTFGTU QH VJG TGCNO QH Saccharomyces cerevisiae (XCP &KLMGP 2001). 6JG IGPGVKE FKXGTUKV[ CPF DKQNQIKECN XGTUCVKNKV[ DGVYGGP FKHHGTGPV [GCUV IGPGTC CPF URGEKGU JCU CNNQYGF HQT VJG WPRTGEGFGPVGF FGXGNQROGPV QH C HWNN UEQTG QH DKQVGEJPQNQIKECN CRRNKECVKQPU. 6JG TQDWUVPGUU QH UGXGTCN PQP-Saccharomyces [GCUVU KU HQT GZCORNG DGKPI GZRNQKVGF KP DKQEQPXGTUKQPU QH CDWPFCPV CITKEWNVWTCN TCY OCVGTKCNU CPF HGGFUVQEMU VQ HWGN CPF EJGOKECNU, VJCV RTGXKQWUN[ YGTG PQV CEEGUUKDNG YKVJ VJG OCLQT DKQVGEJ [GCUV (#TKUVKFQW CPF úGPVKNNk 2000þ öGCVJGTU 2003). /QFGTP [GCUV DKQVGEJPQNQI[ OC[ DG FGHKPGF CU VJG GZRNQKVCVKQP QH [GCUV RJ[UKQNQI[ (VJG NCVG úTQHGUUQT 6QP[ ûQUG, EKVGF KP (9CNMGT 1ÿÿ8)). 6JGTGHQTG, HWNN-UEQTG WVKNK\CVKQP QH [GCUV CPF HWNN KORNGOGPVCVKQP QH VJG [GCUVU KPFWUVTKCN RQVGPVKCN YKNN QPN[ DG RQUUKDNG VJTQWIJ FGVCKNGF CPF VJQTQWIJ WPFGTUVCPFKPI QH [GCUV RJ[UKQNQI[, CPF JGPEG, QH VJG RTQEGUUGU EQPVTQNNKPI [GCUV ITQYVJ CPF OGVCDQNKUO. &WTKPI ITQYVJ KP KPFWUVTKCN UECNG HGTOGPVQTU VJG [GCUVU UGNFQO, KH GXGT GPEQWPVGT RJ[UKQNQIKECN EQPFKVKQPU VJCV EQWNF DG VGTOGF CU PCVWTCN. õP VJG EQPVTCT[, KPFWUVTKCN WUGHWNPGUU CPF CRRNKECVKQP KU NCTIGN[ DGKPI FGVGTOKPGF D[ VJG CDKNKV[ QH VJG [GCUV VQ TGCFKN[ CFCRV VQ XCT[KPI GPXKTQPOGPVCN CPF RJ[UKQNQIKECN EJCNNGPIGU. *GPEG, TQDWUVPGUU CPF IGPGTCN UVTGUU TGUKUVCPEG CTG OCLQT EQPVTQN HCEVQTU FGVGTOKPKPI VJG DKQVGEJPQNQIKECN XCNWG QH VJG [GCUV. 6JGUG EQPVTQN HCEVQTU CTG HQT VJG OQUV RCTV GODGFFGF KP VJG IGPGVKE OCMGWR QH VJG [GCUV CPF UJQWNF DG VJQTQWIJN[ KPXGUVKICVGF VQ GPUWTG WPFGTUVCPFKPI CPF GHHKEKGPV WVKNK\CVKQP. /QTGQXGT, VJQTQWIJ MPQYNGFIG QP VJG IGPGVKE EJCTCEVGTKUVKEU IQXGTPKPI OGVCDQNKE EQPVTQN KU GUUGPVKCN VQ FGXGNQR UVTCKP KORTQXGOGPV UVTCVGIKGU D[ ENCUUKECN IGPGVKEU CRRTQCEJGU (%JCRVGT 2). #NTGCF[ VQ FCVG, VJG GZRNQKVCVKQP QH [GCUVU KP DKQVGEJPQNQI[ JCU CEEWOWNCVGF VQ C FKXGTUG CTTC[ QH CRRNKECVKQPU (øKI. 2, CPF (9CNMGT 1ÿÿ8þ 6CDNG 1 KP %JCRVGT 2)). +V ECP DG UCHGN[ CPVKEKRCVGF VJCV VJKU NKUV YKNN GZVGPF TCRKFN[ KP VJG EQOKPI FGECFG. ;GCUVU ECTT[ C NCTIG RQVGPVKCN HQT VJG WVKNK\CVKQP QH TGPGYCDNG TGUQWTEGU, HQT DKQNQIKECN EQPVTQN (UGG G.I. %JCRVGT 10), HQT FKXGTUG PWVTKVKQPCN KPITGFKGPVU CPF RTQDKQVKEU, CPF KP C YKFG CTGC QH JGCNVJ ECTG KUUWGU. 9KVJ TGICTF VQ VJG NCVVGT, VJG TGEGPVN[ EQORNGVGF IGPQOG UGSWGPEGU QH Saccharomyces cerevisiae (ôQHHGCW GV CN. 1ÿÿ7) CPF Schizosaccharomyces pombe (9QQF GV CN. 2002) EQPVCKP C KORTGUUKXG PWODGT QH IGPGU YKVJ ENGCT VQ GZVGPUKXG JQOQNQI[ VQ IGPGU KPXQNXGF KP FKXGTUG JWOCP VTCKVU CPF FKUGCUGU (&GEQVVKIPKGU CPF ôQHHGCW 1ÿÿ7þ øQWT[ 1ÿÿ7þ ôCTHKPMGN CPF $CKNKU 2002þ
4
*CP FG 9KPFG
(GTOGPVCVKQP KPFWUVTKGU ©PXKTQPOGPV CN VGEJPQNQI[ DKQTGOGFKCVKQP, YCUVG WVKNK\CVKQP, DKQUQTRVKQP QH OGVCNU
DTGYKPI, DCMKPI, DKQ-GVJCPQN, PQXGN RTQEGUUGU, HGTOGPVCVKQP RTQFWEVU
YGCUV $KQVGEJPQNQI[
$KQOGFKECN ÿGUGCTEJ ECPEGT, #+&5, FTWI OGVCDQNKUO, IGPQVQZKEKV[ UETGGPU
(QQF & EJGOKECN +PFWUVTKGU UCXQT[ HNCXQWTU, DCMKPI GP\[OGU, ECTQVGPQKFU, DKQEQPVTQN
*GCNVJ-ECTG KPFWUVTKGU (WPFCOGPVCN $KQNQIKECN ÿGUGCTEJ
RJCTOCEGWVKECNU, DNQQF EQCIWNCVQP HCEVQTU, XCEEKPGU
EGNN DKQNQI[, IGPGVKEU, DKQEJGOKUVT[, OQNGEWNCT DKQNQI[, HWPEVKQPCN IGPQOKEU
(KI. 2. õXGTXKGY QH FKXGTUG CRRNKECVKQP CTGCU QH [GCUV DKQVGEJPQNQI[. #FCRVGF HTQO (9CNMGT, 1ÿÿ8) YKVJ KPRWV HTQO %JCRVGT 2, 6CDNG 1.
1.1.4 5GV-WR QH VJKU DQQM +P VJKU DQQM, YG JCXG CKOGF CV DTKPIKPI VQIGVJGT GZVGPUKXG FGUETKRVKQPU QH DQVJ ENCUUKECN CPF OQNGEWNCT IGPGVKE TGUGCTEJ CPF CRRNKECVKQPU QH XCTKQWU [GCUVU VJCV CTG KORQTVCPV HQT EQPVGORQTCT[ DKQVGEJPQNQI[. +PFGGF, GORJCUKU KU QP KORNKECVKQP CPF CRRNKECVKQP QH [GCUVU KP CITQ-TGNCVGF CPF KPFWUVT[-TGNCVGF DKQVGEJPQNQI[. 9G FGUETKDG GZCORNGU CPF FGVCKNU QP TGUGCTEJ KPVQ CPF CRRNKECVKQP QH (OQNGEWNCT) IGPGVKEU QH [GCUVU CU FKXGTUG CU Saccharomyces, Kluyveromyces, Hansenula, Pichia, CPF Yarrowia. 5EKGPVKHKE MPQYNGFIG CPF KPUKIJV KU FGUETKDGF HQT ÉCPEKGPVÌ DCMGT•U, YKPG, CPF DTGYGT•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ûCVJGT, YG JCXG VTKGF VQ UGV C UVCIG QH KORQTVCPV GZCORNGU QH IGPGVKE OCMGWR, CPCN[UKU, CPF
1 øWPEVKQPCN IGPGVKEU QH KPFWUVTKCN [GCUVUþ QH CPEKGPV UMKNNU CPF OQFGTP CRRNKECVKQPU
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WVKNK\CVKQP QH [GCUVU KP OQFGTP DKQVGEJPQNQI[. +V KU WR VQ VJG KPVGTGUVGF TGCFGT VQ WUG VJG KPHQTOCVKQP CPF GZVGPF KPUKIJV CPF CRRNKECVKQPU VQ QVJGT CTGCU QH DKQVGEJPQNQI[.
1.2 (TQO ENCUUKECN IGPGVKEU VQ OQFGTP IGPQOKEU 6TCFKVKQPCNN[, YKNF V[RG CPF KPFWUVTKCN UVTCKPU QH Saccharomyces cerevisiae CTG HCT NGUU GCUKN[ CEEGUUKDNG HQT IGPGVKE CPCN[UKU CPF OCPKRWNCVKQP VJCP VJGKT NCDQTCVQT[ EQWPVGTRCTVU (%JCRVGT 2). öQY URQTWNCVKQP HTGSWGPE[ CPF XKCDKNKV[ QH URQTGU, KPUVCDKNKV[ QH OCVKPI V[RGU CPF RQQT OCVKPI GHHKEKGPEKGU, CPF VJG CDUGPEG QH GCUKN[ VTCEVCDNG CWZQVTQRJKE OCTMGTU CTG UQOG QH VJG EJCTCEVGTKUVKEU VJCV EQORTQOKUG WUGT HTKGPFNKPGUU QH KPFWUVTKCN [GCUV URGEKGU. 5RGEKGU QVJGT VJCP FQOGUVKECVGF Saccharomyces UQOGVKOGU OC[ QHVGP DG GXGP NGUU CEEGUUKDNG (UGG %JCRVGT 10). *GPEG, UVTCVGIKGU HQT FGFKECVGF UVTCKP KORTQXGOGPV VJTQWIJ ENCUUKECN IGPGVKE CRRTQCEJGU OC[ DG JKPFGTGF UWDUVCPVKCNN[ D[ VJG KPJGTGPV IGPGVKE EQPUVTCKPVU RTGUGPVGF D[ FKHHGTGPV [GCUVU. óGXGTVJGNGUU, CU UVCVGF CDQXG, VJG HWNN KORNGOGPVCVKQP QH VJG RJ[UKQNQIKECN RQVGPVKCN QH FKHHGTGPV [GCUVU DGIKPU YKVJ C VJQTQWIJ WPFGTUVCPFKPI QH VJGKT IGPGVKE DCEMITQWPF CPF FGNKPGCVKQP QH KORQTVCPV RJ[UKQNQIKECN EQPVTQN OGEJCPKUOU. &GVCKNGF IGPGVKE CPCN[UKU KU TGSWKTGF VQ EJCTCEVGTK\G TGNGXCPV CPF GHHKEKGPV VCTIGVU HQT UVTCKP KORTQXGOGPV CPF HQT FKTGEVGF UETGGPKPI RTQEGFWTGU KP UVTCKP UGNGEVKQP RTQITCOU. 9KVJ VJG EQORNGVKQP QH VJG IGPQOG UGSWGPEKPI RTQLGEV QH Saccharomyces cerevisiae (ôQHHGCW GV CN. 1ÿÿ7) CNUQ MPQYP CU DCMGT•U, YKPG, QT DTGYGT•U [GCUV (UGG %JCRVGTU 3, 4 CPF ý), YG CTG PQY HCEKPI VJG EJCNNGPIG QH WPEQXGTKPI VJG HWNN ÂJKFFGP RQVGPVKCN• QH VJKU FQOGUVKECVGF [GCUV, CPF QH OCP[ QVJGT [GCUVU KP KVU VTCKN. # OCLQT VTKCN NKGU KP DQVJ TGXGCNKPI RJ[UKQNQIKECN EQPVTQN OGEJCPKUOU QH VJG [GCUVU (CPF QH EQWTUG, QVJGT OKETQ-QTICPKUOU) CPF KP CUUKIPKPI HWPEVKQPCN TQNGU VQ CNN RTQVGKPU GPEQFGF D[ VJG IGPQOGU VJCV CTG PQY TCRKFN[ DGEQOKPI CXCKNCDNG. # RTQOKUKPI UVTCVGI[ VQ EQPXGTV EWTTGPV ÉDNCEM DQZÌ CRRTQCEJGU QH [GCUV RJ[UKQNQI[ VQ HWNN[ EJCTCEVGTK\GF ÉYJKVG DQZÌ MPQYNGFIG NKGU KP VJG KPVGITCVKQP QH EGNN DKQNQI[, RJ[UKQNQI[, OQNGEWNCT DKQNQI[, IGPGVKEU CPF DKQEJGOKUVT[, EQODKPGF KP PQXGN HWPEVKQPCN IGPQOKEU CRRTQCEJGU (øKI. 3). 6JGUG CNNQY HQT VJG KPXGUVKICVKQP QH HWPEVKQP CPF KORCEV QH OCP[ IGPGU CPF RTQVGKPU KP RCTCNNGN (%JCRVGT 11). #NUQ JGTG, VJG RKQPGGTKPI UVWFKGU GORNQ[KPI Saccharomyces cerevisiae JCXG KP TGEGPV [GCTU UJQYP VJG GPQTOQWU RQVGPVKCN QH WUKPI VJGUG VQQNU, CPCN[\KPI QP C IGPQOG-YKFG UECNG IGPG GZRTGUUKQP RTQHKNGU, IGPGVKE KPVGTCEVKQPU, RTQVGKP GZRTGUUKQP CPF NQECNK\CVKQP, CPF DKQEJGOKECN TGCEVKQPU CPF CEVKXKVKGU (TGXKGYGF KP (òWG CPF 9KP\GNGT 2002þ /CTVKP CPF &TWDKP 2003)). +PVGITCVKQP CPF EQODKPCVKQP QH QWT MPQYNGFIG QP [GCUV IGPGVKEU CPF RJ[UKQNQI[ YKNN GPCDNG UWEEGUUHWN UEKGPVKHKE CPF EQOOGTEKCN GZRNQKVCVKQP QH [GCUV KP DKQVGEJPQNQI[.
ü
*CP FG 9KPFG
Enzymes, HPLC, NMR
Bio-informatics
Metabolome Proteome Transcriptome
2D gels
Genome DNA chips (KI. 3. øTQO ÉDNCEM DQZÌ RJ[UKQNQI[ VQ ÉYJKVG DQZÌ MPQYNGFIG. 9KVJ KPVGITCVKXG CRRTQCEJGU WUKPI OQNGEWNCT IGPGVKEU CPF PQXGN HWPEVKQPCN IGPQOKEU VGEJPQNQI[, YG YKNN DG CDNG VQ HKNN VJG EWTTGPV MPQYNGFIG ICR QP VJG IGPGVKE DCUKU QH RJ[UKQNQIKECN EQPVTQN OGEJCPKUOU QRGTCVKPI KP [GCUV. øQT C OQTG FGVCKNGF GZRNCPCVKQP, UGG VJG VGZV.
1.3 +ORTQXKPI CPEKGPV UMKNNU: Saccharomyces cerevisiae KP HQQF CPF DGXGTCIG 1.3.1 $CMGT•U [GCUV #U FGUETKDGF CDQXG, [GCUVU JCXG C NQPIUVCPFKPI JKUVQT[ KP DTGCF OCMKPI. 6JGKT OCKP HWPEVKQP KP VJG DTGCF OCMKPI RTQEGUU KU VQ TGCFKN[ HGTOGPV UWICTU VJCV CTG CXCKNCDNG KP VJG HNQWT QH VJG FQWIJ QT VJCV CTG DGKPI CFFGF VQ VJG FQWIJ. #U C TGUWNV QH GHHKEKGPV HGTOGPVCVKQP, VJG [GCUV RTQFWEGU %õ2 CPF GVJCPQNþ VJG ECTDQP FKQZKFG KU VTCRRGF YKVJKP VJG INWVGP OCVTKZ QH VJG FQWIJ CPF ECWUGU VJG NGCXGPKPI QT TKUKPI, YJGTGCU VJG GVJCPQN EQPVTKDWVGU VQ HNCXQWT HQTOCVKQP, KP EQPEGTV YKVJ QVJGT XQNCVKNG EQORQWPFU CPF HNCXQWT RTGEWTUQTU VJCV HQTO FWTKPI VJG CPCGTQDKE HGTOGPVCVKQP RTQEGUU. 6JG RJ[UKQNQIKECN FGOCPF RWV VQFC[ QP DCMGT•U [GCUV DGVYGGP RTQFWEVKQP CPF CRRNKECVKQP EQPUVKVWVGU CP CRRCTGPV RCTCFQZ (øKI. 4). +PFGGF, UWICT-NKOKVGF TGURKTQHGTOGPVCVKXG HGF DCVEJ EWNVKXCVKQP KU OGCPV VQ [KGNF C [GCUV RTQFWEV VJCV JCU FGXGNQRGF C JKIJ HGTOGPVCVKXG ECRCEKV[ CNVJQWIJ VJKU KU PQV TGSWKTGF FWTKPI VJG RTQFWEVKQP RJCUG. 6JKU JKIJ ICUUKPI ECRCEKV[ UWDUGSWGPVN[ KU TGETWKVGF KP VJG CRRNKECVKQP RJCUG KP VJG FQWIJ, WPFGT CPCGTQDKE, UWICT GZEGUU EQPFKVKQPU. 6JG TGSWKTGF RJ[UKQNQIKECN HNGZKDKNKV[ KU V[RKECN HQT DCMGT•U [GCUV. #V VJG UCOG VKOG, KV KU UWDLGEV VQ IGPGVKE UVTCKP KORTQXGOGPV RTQITCOU YKVJ UGXGTCN KORQTVCPV DCMGT•U [GCUV RTQ-
1 øWPEVKQPCN IGPGVKEU QH KPFWUVTKCN [GCUVUþ QH CPEKGPV UMKNNU CPF OQFGTP CRRNKECVKQPU
Application
Production
(GTOGPVCVKQP - %12
4GURKTCVKQP - $KQOCUU LQw INWEQUG #GTQDKE
*KIJ OCNVQUG/INWEQUG #PCGTQDKE 57û#ÿ
57û#ÿ ú þ2
(GTOGPVCVKXG %#ü#%+6Y
EGNNU
7
%þ2 ý©VJCPQN)
(GTOGPVCVKXG #%6+8+6Y
EGNNU
%þ2 ý©VJCPQN)
(KI. 4. 6JG RCTCFQZ QH DCMGT•U [GCUV RTQFWEVKQP CPF CRRNKECVKQP. $CMGT•U [GCUV KU RTQFWEGF KP UWICT-NKOKVGF, TGURKTQ-HGTOGPVCVKXG HGF DCVEJ EWNVKXCVKQPU, CPF WPFGT VJGUG EQPFKVKQPU UJQWNF FGXGNQR VJG RQVGPVKCN VQ TGCFKN[ HGTOGPV GZEGUU UWICT VQ %õ2 (CPF GVJCPQN) KP UWDUGSWGPV FQWIJ CRRNKECVKQP. 6JWU, WPFGT TGURKTCVQT[ RJ[UKQNQIKECN EQPFKVKQPU YJGTG OGVCDQNKE HNWZ KU VQYCTF EGNN ITQYVJ CPF DKQOCUU RTQFWEVKQP, YG GZRGEV VJG [GCUV VQ FGXGNQR C HWNNUEQTG HGTOGPVCVKXG ECRCEKV[. øQT OQTG FGVCKNU, UGG VGZV CPF %JCRVGT 3.
FWEGTU, VQ OGGV KPETGCUKPI FGOCPFU HQT HNGZKDKNKV[ WPFGT C YKFGPKPI TCPIG QH FKXGTUG OCTMGV CRRNKECVKQPU. +P %JCRVGT 3 QH VJKU DQQM, YG GNCDQTCVG QP HWPFCOGPVCN CPF CRRNKGF CURGEVU QH DCMGT•U [GCUV IGPGVKEU VJCV EQPVTKDWVG VQ QWT EWTTGPV WPFGTUVCPFKPI QH VJG URGEKHKE EQPUVTCKPVU KORQUGF QP DCMGT•U [GCUV RJ[UKQNQI[ KP VGTOU QH RTQFWEVKQP CPF CRRNKECVKQP. 6JG [GCUV ICUUKPI TCVG, K.G. VJG TCVG QH %õ2 RTQFWEVKQP KPUKFG VJG FQWIJ KU ETKVKECN KP DCMKPI VGEJPQNQI[, CPF OQTG KORQTVCPVN[ UQ YJGP EQORCTKPI VJG FKHHGTGPV FQWIJ CRRNKECVKQPU RTGUGPVKPI NCTIGN[ XCT[KPI RJ[UKQNQIKECN EQPFKVKQPU VQ VJG [GCUV (#VVHKGNF 1ÿÿ7). ùPQYNGFIG QP CPF EQPVTQN QH VJG FKXGTUG UVTGUU TGURQPUG OGEJCPKUOU VJG [GCUV OC[ TGETWKV WPFGT FKHHGTGPV CRRNKECVKQP EQPFKVKQPU UWEJ CU UYGGV FQWIJ, UQWT FQWIJ, HTQ\GP FQWIJ GVE. YKNN DQQUV UVTCVGIKGU QP UVTCKP KORTQXGOGPV CPF GPJCPEG GHHKEKGPV WVKNK\CVKQP QH VJG DCMGT•U [GCUV HWNN RJ[UKQNQIKECN RQVGPVKCN. 1.3.2 9KPG [GCUV #U DTKGHN[ OGPVKQPGF CDQXG, VJG CTV QH YKPG OCMKPI RTQDCDN[ RTGUGPVU VJG QNFGUV VGEJPQNQIKECN EWNVKXCVKQP QH [GCUVU D[ OCP. õPN[ FWTKPI VJG NCUV EGPVWT[, C UEKGPVKHKE DCUKU QH YKPG OCMKPI JCU ITCFWCNN[ ITQYP, EWNOKPCVKPI KP FGVCKNGF KPXGUVKICVKQPU KPVQ VJG DKQNQIKECN CPF RJ[UKQNQIKECN FKXGTUKV[ QH [GCUV URGEKGU CUUQEKCVGF YKVJ YKPG OCMKPI (úTGVQTKWU 2000). #PCN[UKU QH VJG URGEKHKE IGPGVKE EQPUVKVWVKQP
8
*CP FG 9KPFG
CPF KFGPVKHKECVKQP QH IGPGVKE VCTIGVU HQT EQPVKPWGF KORTQXGOGPV QH YKPG [GCUVU CTG RTGUGPVN[ KORQTVCPV FTKXKPI HQTEGU HQT KPPQXCVKQPU KP YKPG OCMKPI. %JCRVGT 4 RTGUGPVU CP GZVGPUKXG CPF DGCWVKHWN QXGTXKGY QH EWTTGPV MPQYNGFIG CPF WPFGTUVCPFKPI QP YKPG [GCUV IGPGVKEU CPF EQPVTQN OGEJCPKUOU IQXGTPKPI VJG [GCUVU TGSWKTGF RJ[UKQNQIKECN EQPUVKVWVKQP FWTKPI YKPG HGTOGPVCVKQP. 5GXGTCN RJ[UKQNQIKECN VCTIGVU CPF UGNGEVKQP UVTCVGIKGU HQT VJG IGPGVKE VCKNQTKPI CPF KORTQXGOGPV QH YKPG [GCUV CTG FKUEWUUGF. 5RGEKCN CVVGPVKQP KU RCKF VQ VJG TQNG VJCV EWUVQOKUGF UVCTVGT [GCUV UVTCKPU OC[ RNC[ KP KORTQXKPI HGTOGPVCVKQP, RTQEGUUKPI CPF DKQRTGUGTXCVKQP QH YKPGU, CPF VJGKT ECRCEKV[ VQ GPJCPEG YJQNGUQOGPGUU CPF UGPUQT[ SWCNKV[. 1.3.3 $TGwGT•U [GCUV 6JG CTV QH DGGT DTGYKPI KU CPEKGPV, CPF Saccharomyces [GCUV OC[ JCXG RNC[GF C EGPVTCN TQNG KP VJG DTGYKPI RTQEGUU HTQO VJG UVCTV. öKMG KP YKPG OCMKPI, C UEKGPVKHKE DCUKU HQT DGGT DTGYKPI JCU QPN[ ITQYP YKVJ CP KPETGCUKPI URGGF FWTKPI VJG NCUV EQWRNG QH FGECFGU, KPENWFKPI IGPGVKE CPF RJ[UKQNQIKECN EJCTCEVGTK\CVKQP QH DTGYGT•U [GCUVU. 6JG RTQFWEVKQP QH DGGT KPXQNXGU UGXGTCN FKHHGTGPV KORQTVCPV UVGRU, QH YJKEJ QPN[ VJG NCVVGT FGRGPF QP [GCUV. 6JG EJCTCEVGTKUVKEU QH VJG [GCUV CTG, JQYGXGT, GUUGPVKCNN[ FGEKUKXG HQT VJG VJTQWIJRWV QH VJG YJQNG RTQEGUU, CPF HQT VJG SWCNKV[ CPF RTQRGTVKGU QH VJG GPF RTQFWEV. /QTGQXGT, DGGT RTQFWEKPI HGTOGPVCVKQPU CTG WPKSWG YKVJKP VJG DGXGTCIG KPFWUVT[, UKPEG VJG [GCUV KU PQV FKUECTFGF CHVGT WUCIG, DWV OCKPVCKPGF CPF TGWUGF C PWODGT QH VKOGU (úQYGNN GV CN. 2003). 6JKU KORNKECVGU VJCV CIGKPI-TGNCVGF CNVGTCVKQPU KP [GCUV RJ[UKQNQI[, OQTRJQNQI[ CPF QXGTCNN IGPG GZRTGUUKQP RNC[ CP KORQTVCPV TQNG KP VJG [GCUV RGTHQTOCPEG FWTKPI DGGT HGTOGPVCVKQP. 6JKU TGEGPV KPUKIJV KPVQ VJG KORCEV QH EGNN CIGKPI JCU CFFGF CP GZVTC UEQRG QP IGPGVKE CPF RJ[UKQNQIKECN KPXGUVKICVKQPU QH DTGYGT•U [GCUV. +P %JCRVGT ý, YG FGUETKDG EWTTGPV MPQYNGFIG CPF WPFGTUVCPFKPI QH DTGYKPI [GCUV RJ[NQIGP[, IGPGVKEU, CPF RJ[UKQNQI[. ôGPGVKE VGEJPQNQIKGU VJCV CTG CXCKNCDNG HQT UVTCKP KORTQXGOGPV CTG FKUEWUUGF, CU CTG OCLQT CTGCU HQT VJG KORTQXGOGPV QH VJG DTGYKPI [GCUV DGJCXKQWT CPF RGTHQTOCPEG. %JCTCEVGTKUVKE CPF KORQTVCPV RTQRGTVKGU CTG RCTVN[ QXGTNCRRKPI VJQUG QH YKPG [GCUV, CPF KPENWFG WRVCMG QH ECTDQJ[FTCVGU, COKPQ CEKFU CPF KQPU, RTKOCT[ OGVCDQNKUO (VWTPKPI ECTDQJ[FTCVGU KPVQ GPGTI[ CPF CNEQJQN), UGEQPFCT[ OGVCDQNKUO (HQTOCVKQP QH CTQOC EQORQWPFU), UGFKOGPVCVKQP EJCTCEVGTKUVKEU CPF UVTGUU TGURQPUG, XKVCNKV[ CPF XKCDKNKV[.
1.4 þH CRRNGU CPF RGCTU: PGw HQQF [GCUV +P RCTCNNGN VQ Saccharomyces cerevisiae, UGXGTCN QVJGT [GCUVU JCXG HQWPF, QT CTG DGIKPPKPI VQ HKPF VJGKT YC[ VQ CRRNKECVKQPU KP HQQF DKQVGEJPQNQI[. #P QDXKQWU RTGTGSWKUKVG KU VJCV VJQUG [GCUVU NKMG Saccharomyces JCXG CP CRRTQXGF ôû#5 (IGPGTCNN[ TGICTFGF CU UCHG) UVCVWU HQT WVKNK\CVKQP CPF KORNGOGPVCVKQP QH HQQF CFFKVKXGU ($QPGMCOR CPF õQUVGTQO 1ÿÿ4). 5KPEG VJGKT IGPGVKE CPF RJ[UKQNQIKECN EJCTCEVGT-
1 øWPEVKQPCN IGPGVKEU QH KPFWUVTKCN [GCUVUþ QH CPEKGPV UMKNNU CPF OQFGTP CRRNKECVKQPU
ÿ
KUVKEU CTG FKHHGTGPV HTQO VJQUG QH Saccharomyces NKMG CRRNGU HTQO RGCTU, VJGUG PQXGN HQQF [GCUVU RTQOKUG VQ YKFGP VJG RCNGVVG QH RQUUKDKNKVKGU HQT EGTVCKP URGEKHKE CPF PGY DKQVGEJPQNQIKECN CRRNKECVKQPU. 1.4.1 Kluyveromyces lactis Kluyveromyces lactis YCU VJG HKTUV [GCUV HQNNQYKPI Saccharomyces cerevisiae VJCV QDVCKPGF VJG ôû#5 UVCVWU HQT HQQF CFFKVKXG WUCIG ($QPGMCOR CPF õQUVGTQO 1ÿÿ4). óQYCFC[U, VJKU [GCUV KU RTQDCDN[ VJG DGUV-GUVCDNKUJGF PQP-Saccharomyces [GCUV KP DKQVGEJPQNQIKECN TGUGCTEJ CPF CRRNKECVKQP. Kluyveromyces JCU RTQXGP VQ DG C HCT DGVVGT JQUV HQT TGEQODKPCPV RTQVGKP RTQFWEVKQP VJCP Saccharomyces, PQV KP VJG NGCUV DGECWUG QH OCTMGF FKHHGTGPEGU KP VJG EQPVTQN QH ECTDQP WVKNK\CVKQP CPF VJG CDUGPEG QH VJG UQ-ECNNGF %TCDVTGG GHHGEV (FG &GMGP 1ÿüü) QH QDNKICVG CNEQJQNKE CGTQDKE HGTOGPVCVKQP WPFGT UWICT GZEGUU. 6JKU Saccharomyces-URGEKHKE EJCTCEVGTKUVKE KU JKIJN[ WUGHWN HQT HGTOGPVCVKXG CRRNKECVKQPU KP DCMGT[, YKPG OCMKPI CPF DGGT DTGYKPI, DWV KU FGNGVGTKQWU HQT JKIJ RTQFWEV [KGNFU KP RTQVGKP RTQFWEVKQP. *GPEG, Kluyveromyces lactis JCU DGGP GORNQ[GF KP VJG RTQFWEVKQP QH UGXGTCN HQQF GP\[OGU, UWEJ CU NCEVCUG CPF EJ[OQUKP (XCP FGP $GTI GV CN. 1ÿÿ0þ 5YKPMGNU GV CN. 1ÿÿ3). ùKNNGT UVTCKPU QH Kluyveromyces JCXG DGGP WUGF KP DKQEQPVTQN QH ITCRG OWUV CPF UKNCIG HGTOGPVCVKQPU, CPF URGEKHKE UVTCKPU JCXG DGGP GPIKPGGTGF VQ RTQFWEG NCEVKE CEKF. %JCRVGT ü RTGUGPVU C FGVCKNGF QXGTXKGY QH EWTTGPV WPFGTUVCPFKPI QH Kluyveromyces IGPGVKE OCMGWR CPF EQPUGSWGPVN[, KVU RJ[UKQNQIKECN EQPVTQN TGRGTVQKTG. 9KVJ KVU IGPQOG UGSWGPEG CDQWV VQ DG HWNN[ WPTCXGNNGF ($QNQVKP-øWMWJCTC GV CN. 2000) VJKU [GCUV KU CU GCUKN[ CEEGUUKDNG HQT IGPGVKE CPCN[UKU CPF OCPKRWNCVKQP CU KVU FKUVCPV TGNCVKXG Saccharomyces, GORJCUKUKPI KVU KORQTVCPV TQNG KP OQFGTP DKQVGEJPQNQI[. 1.4.2 Torulaspora delbrueckii õTKIKPCNN[ EQPUKFGTGF CU C URQKNCIG [GCUV (5EJ·NNGT GV CN. 2001), Torulaspora delbrueckii JCU DGEQOG CP KPVTKIWKPI GZCORNG QH PQXGN [GCUVU EQOKPI QH CIG KP OQFGTP DKQVGEJPQNQI[. 5VTCKPU QH VJKU [GCUV YGTG TGEQIPK\GF HQT VJGKT GZVTGOG QUOQVKE CPF HTGG\G VQNGTCPEG CPF TGUKUVCPEG VQ UGXGTCN QVJGT V[RGU QH RJ[UKQNQIKECN UVTGUUGU (#NOGKFC CPF úCKU 1ÿÿü). 6JGUG RTQRGTVKGU JCXG OCFG VJKU [GCUV JKIJN[ CVVTCEVKXG CU C PQXGN DCMGT•U [GCUV HQT CRRNKECVKQP KP JKIJ UWICT FQWIJU (QUOQVKE UVTGUU) CPF HTQ\GP FQWIJU (HTGG\G UVTGUU). $GKPI C PQXGN DKQVGEJPQNQIKECNN[ TGNGXCPV [GCUV, IGPGVKE CPCN[UKU CPF IGPGVKE VQQNU CTG QPN[ DGIKPPKPI VQ FGXGNQR. +P %JCRVGT 3, VJKU RTQOKUKPI GZCORNG QH C PGY DKQVGEJPQNQIKECN [GCUV KU FGUETKDGF KP EQPUKFGTCDNG FGVCKN.
10
*CP FG 9KPFG
1.ù YGCUV CU OQFGN U[UVGO 1.ù.1 Saccharomyces [GCUV KP UKIPCN VTCPUFWEVKQP CPF EGNN wCNN DKQU[PVJGUKU ôKXGP KVU UVCVWU CU C YKFGN[ EWNVKXCVGF [GCUV CPF CV VJG UCOG VKOG DGKPI IGPGVKECNN[ YGNN CEEGUUKDNG, Saccharomyces cerevisiae JCU FWTKPI VJG NCUV VYQ FGECFGU UGTXGF CU CP KFGCN GWMCT[QVKE OQFGN U[UVGO HQT VJG WPTCXGNNKPI QH VJG IGPGVKE DCUKU QH OCP[ EGNNWNCT CPF RJ[UKQNQIKECN EQPVTQN OGEJCPKUOU. 5KIPCN VTCPUFWEVKQP RCVJYC[U CPF PGVYQTMU VJCV CTG QRGTCVKPI KP TGURQPUG VQ XCTKQWU GZVTCEGNNWNCT UVKOWNK, PWVTKGPV CXCKNCDKNKV[, CPF GPXKTQPOGPVCN EJCPIGU JCXG DGGP FGNKPGCVGF KP FGVCKN (9KPFGTKEMZ GV CN. 2002). 6JGUG UKIPCNNKPI ECUECFGU YKVJQWV GZEGRVKQP JCXG VJGKT EQWPVGTRCTVU KP JKIJGT GWMCT[QVGU, VJG WPTCXGNNKPI QH YJKEJ JCU DGGP NCTIGN[ HCEKNKVCVGF D[ MPQYNGFIG FGTKXGF HTQO VJG [GCUV GZCORNGU. 6JKU HQWPFKPI TQNG QH Saccharomyces cerevisiae CPF, HQT C NKVVNG NGUU VKOG DWV PQV NGUU KORQTVCPVN[, QH Schizosaccharomyces pombe HQT QWT WPFGTUVCPFKPI QH VJG OGEJCPKUOU TGIWNCVKPI EGNN ITQYVJ CPF FKXKUKQP JCU TGEGPVN[ DGGP YKFGN[ CEMPQYNGFIGF YJGP NGCFKPI TGUGCTEJGTU CPF ÂHQWPFKPI HCVJGTU• QH [GCUV UKIPCN VTCPUFWEVKQP TGUGCTEJ YQP VJG óQDGN úTK\G KP RJ[UKQNQI[ CPF OGFKEKPG (*CTVYGNN 2002þ óWTUG 2002). ©ZCORNGU QH TGUGCTEJ CPF CRRNKECVKQP QH VJG IGPGVKEU QH UKIPCN VTCPUFWEVKQP CPF RJ[UKQNQIKECN EQPVTQN OGEJCPKUOU KP Saccharomyces cerevisiae CTG FGUETKDGF KP EQPUKFGTCDNG FGVCKN KP %JCRVGTU 3 (DCMGT•U [GCUV), 4 (YKPG [GCUV) CPF ý (DTGYGT•U [GCUV). 6JG IGPGVKE DCUKU HQT URGEKHKE EJCTCEVGTKUVKEU QH KORQTVCPV OGVCDQNKE TGURQPUGU KP GCEJ QH VJGUG FKHHGTGPV DKQVGEJPQNQIKECN CRRNKECVKQPU HWTVJGT KNNWUVTCVGU VJG RJ[UKQNQIKECN HNGZKDKNKV[ CUUQEKCVGF YKVJ VJKU [GCUV IGPWU. %JCRVGT ÿ FGUETKDGU CPQVJGT KORQTVCPV KUUWG QH [GCUV KP DKQVGEJPQNQI[, CPF VJG TGNCVKQP YKVJ GPXKTQPOGPVCN UKIPCN VTCPUFWEVKQP TGUGCTEJ CPF OCPKRWNCVKQP. ;GCUVU (CPF HWPIK) CTG C OCLQT UQWTEG QH HQQF URQKNCIG, ECWUKPI EQPUKFGTCDNG GEQPQOKE NQUUGU. +PXGUVKICVKQPU QXGT VJG NCUV [GCTU KPVQ VJG IGPGVKE DCUGU QH [GCUV OGEJCPKUOU EQPVTQNNKPI EGNN UWTXKXCN, UVTGUU TGUKUVCPEG, CPF EGNN YCNN EQPUVKVWVKQP JCU EQPVTKDWVGF NCTIGN[ VQ VJG FGXGNQROGPV QH CPVK-[GCUV CPF CPVK-HWPICN UVTCVGIKGU YJKEJ CTG EWTTGPVN[ DGKPI CRRNKGF CPF HWTVJGT FGXGNQRGF. 1.ù.2 Kluyveromyces lactis #U FGUETKDGF CDQXG Kluyveromyces lactis DGKPI FKUVCPVN[ TGNCVGF VQ Saccharomyces URR., JCU RTQXGP CPQVJGT KORQTVCPV OQFGN U[UVGO HQT UKIPCN VTCPUFWEVKQP PGVYQTMU TGIWNCVKPI EGNN ITQYVJ CPF RJ[UKQNQI[ KP TGURQPUG VQ GPXKTQPOGPVCN EJCPIGU. #U FGUETKDGF KP %JCRVGT ü, EQPVTQN OGEJCPKUOU QH KPVGTOGFKCT[ OGVCDQNKUO CTG RCTVN[ EQORCTCDNG, CPF RCTVN[ FKHHGTGPV HTQO VJQUG KP Saccharomyces [GCUV. *GPEG, C VJQTQWIJ KPXGUVKICVKQP QH VJG IGPGVKE DCUKU WPFGTN[KPI VJGUG FKHHGTGPEGU JCU EQPVTKDWVGF VQ QWT WPFGTUVCPFKPI QH VJG HCEVQTU FGVGTOKPKPI URGEKGUURGEKHKEKV[ CPF HNGZKDKNKV[ KP CFCRVCVKQP VQ FKHHGTGPV ITQYVJ EQPFKVKQPU. +P CFFKVKQP, UWEJ MPQYNGFIG KU ETWEKCN VQ EQOG VQ CP QRVKOCN WVKNK\CVKQP QH [GCUV URGEKGU KP URGEKHKE DKQVGEJPQNQIKECN CRRNKECVKQPU.
1 øWPEVKQPCN IGPGVKEU QH KPFWUVTKCN [GCUVUþ QH CPEKGPV UMKNNU CPF OQFGTP CRRNKECVKQPU
11
1.ù.3 Hansenula, Pichia, CPF Yarrowia [GCUVU 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris, CPF VJG J[FTQECTDQP WVKNK\KPI [GCUV Yarrowia lipolytica RTGUGPV XGT[ PKEG GZCORNGU QH PQP-Saccharomyces [GCUVU VJCV JCXG HQWPF VJGKT YC[ KP KORQTVCPV PKEJGU QH OKETQDKCN DKQVGEJPQNQI[ (*QNNGPDGTI CPF ôGNNKUUGP 1ÿÿ7þ /CF\CM GV CN. 2003). 6JGUG [GCUVU CTG VCZQPQOKECNN[ FKUVCPV HTQO Saccharomyces URR. CPF JCXG FGXGNQRGF URGEKHKE OGVCDQNKE CPF RJ[UKQNQIKECN EJCTCEVGTKUVKEU VJCV JCXG RTQXGP VQ DG KPXCNWCDNG HQT URGEKHKE DKQVGEJPQNQIKECN CRRNKECVKQPU. /QTGQXGT, VJGKT URGEKHKE RJ[UKQNQIKECN EQPUVKVWVKQP JCU OCFG VJGUG [GCUVU KORQTVCPV OQFGN QTICPKUOU HQT VJG WPTCXGNNKPI QH GWMCT[QVKE TGIWNCVQT[ RCVJYC[U KPXQNXGF KP RTQVGKP RTQFWEVKQP, CEEWOWNCVKQP, CPF UGETGVKQP. %JCRVGTU 7 CPF 8 RTGUGPV CP GZVGPUKXG CPF FGVCKNGF QXGTXKGY QH EWTTGPV MPQYNGFIG QP VJG IGPGVKE WPKSWGPGUU CPF EQPUVKVWVKQP QH VJGUG [GCUVU, YJKEJ HQTOU VJG DCUKU HQT VJGKT WVKNK\CVKQP DQVJ CU KORQTVCPV OQFGN U[UVGOU CPF CU DKQVEJPQNQIKECN VQQN QTICPKUOU.
1.ø 0QXGN DKQVGEJPQNQIKECN VQQNU CPF CRRNKECVKQPU 1.ø.1 YGCUV KP DKQEQPVTQN õH QNF, VJG RTQFWEVKQP QH GVJCPQN D[ Saccharomyces [GCUV KP G.I. YKPG HGTOGPVCVKQP JCU DGGP TGEQIPK\GF VQ KPHGT CPVKOKETQDKCN CEVKXKV[, KPJKDKVKPI VJG RTQNKHGTCVKQP QH ÉWPYCPVGFÌ DCEVGTKC CPF HWPIK CV KPETGCUKPI GVJCPQN EQPEGPVTCVKQPU (úTGVQTKWU 2000). /QTG TGEGPVN[, PQXGN OGEJCPKUOU QH CPVKOKETQDKCN CEVKXKV[ JCXG DGGP HQWPF YKVJ C XCTKGV[ QH FKHHGTGPV [GCUVU. 5GXGTCN QH VJGUG [GCUV URGEKGU CTG KPETGCUKPIN[ DGKPI WUGF KP DKQEQPVTQN QH FGNGVGTKQWU HWPIK (KPENWFKPI [GCUVU) KP XCTKQWU CTGCU QH DKQVGEJPQNQI[ KPENWFKPI CITKEWNVWTG, RQUVJCTXGUV VGEJPQNQI[, HQTGUV KPFWUVT[, CPF HQQF CRRNKECVKQPU. %JCRVGT 10 RTGUGPVU CP GZVGPUKXG QXGTXKGY QH VJG EWTTGPV UVCVWU QH IGPGVKE CPCN[UKU CPF KFGPVKHKECVKQP QH VJG YKFG XCTKGV[ QH EWTTGPVN[ WUGF CPF RQVGPVKCNN[ RTQOKUKPI [GCUV URGEKGU KP DKQ EQPVTQN, CPF VJGKT FKHHGTGPV OQFGU QH CEVKQP. #NVJQWIJ TGRTGUGPVKPI C TGNCVKXGN[ [QWPI CTGC QH [GCUV VGEJPQNQI[, KPHQTOCVKQP QP CPF KPUKIJV KPVQ FKXGTUG DKQEQPVTQN OGEJCPKUOU CTG TCRKFN[ ITQYKPI. 9KVJ VJG KPRWV QH OQFGTP IGPGVKEU CPF IGPQOKEU OGVJQFQNQIKGU, VJG CRRNKECVKQP QH [GCUVU KP DKQEQPVTQN YKNN UQQP EQPUVKVWVG CP KORQTVCPV UGEVQT KP OQFGTP DKQVGEJPQNQI[. 1.ø.2 ÷GVCDQNKE RCVJwC[ GPIKPGGTKPI +P DKQVGEJPQNQI[ KPFWUVT[, VJGTG KU C EQPVKPWKPI YKUJ VQ KORTQXG OKETQDKCN HGTOGPVCVKQP CPF RTQFWEVKQP RTQEGUUGU. 6JGTGHQTG, KP VJG NCUV FGECFG IGPGVKE OCPKRWNCVKQP QH OKETQDKCN OGVCDQNKE TQWVGU VGTOGF CU OGVCDQNKE RCVJYC[ GPIKPGGTKPI (/ú©) JCU KPETGCUKPIN[ DGGP GORNQ[GF KP UVTCKP CPF RTQEGUU KORTQXGOGPV RTQITCOU, KPENWFKPI UVTCVGIKGU WUKPI [GCUV (óKGNUGP 2001). /GVCDQNKE GPIKPGGTKPI KU FGHKPGF CU VJG TCVKQPCN KORTQXGOGPV QH EGNNWNCT RTQEGUUGU VJTQWIJ
12
*CP FG 9KPFG
FGHKPGF CU VJG TCVKQPCN KORTQXGOGPV QH EGNNWNCT RTQEGUUGU VJTQWIJ VJG KPVTQFWEVKQP QH FKTGEVGF EJCPIGU KP VJG IGPGVKE DCEMITQWPF QH VJG QTICPKUO, XKC IGPGVKE GPIKPGGTKPI ($CKNG[ 1ÿÿ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ú© QH KPFWUVTKCN [GCUV. 6JG TGUWNVKPI EKTEWNCT RTQEGUU QH GPIKPGGTKPI [GCUV UVTCKPU HQT QRVKOK\GF QT PGY RTQFWEV HQTOCVKQP, VJTQWIJ INQDCN KPXGUVKICVKQP CPF OCPKRWNCVKQP QH IGPGVKE HCEVQTU KPXQNXGF JCU CNTGCF[ KP C PWODGT QH ECUGU [KGNFGF RQUKVKXG TGUWNVU (øKIWTG ý). Saccharomyces cerevisiae JCU HQT GZCORNG DGGP UWEEGUUHWNN[ GPIKPGGTGF, CPF QRVKOK\GF HQT VJG RTQFWEVKQP QH UGXGTCN PQXGN NKRKFU, YJGP ITQYP QP HCVV[ CEKFU CU UQNG ECTDQP UQWTEG (&[GT GV CN., 2002). 6JKU OC[ RTGUGPV C XCNWCDNG VQQN HQT VJG DKQEQPXGTUKQP QH EJGCR QKNU KPVQ JKIJ-XCNWG NKRKF RTQFWEVU. ûGEGPVN[, CPQVJGT DGCWVKHWN GZCORNG QH OGVCDQNKE RCVJYC[ GPIKPGGTKPI KP DCMGT•U [GCUV YCU RWDNKUJGF (5\E\GDCTC GV CN., 2003). #P CTVKHKEKCN, UGNH-UWHHKEKGPV DKQU[PVJGUKU RCVJYC[ KPXQNXKPI PQ NGUU VJCP 13 JGVGTQNQIQWU GPIKPGGTGF IGPGU YCU CUUGODNGF CPF GZRTGUUGF
(WPEVKQPCN ûGPQOKEU
From products to genes
/ÿ©
From genes to products (KI. 5. 5VTCKP CPF RTQEGUU KORTQXGOGPV VJTQWIJ OGVCDQNKE RCVJYC[ GPIKPGGTKPI (/ú©) KU C EKTEWNCT RTQEGUU. +ORNGOGPVCVKQP QH HWPEVKQPCN IGPQOKEU VGEJPQNQI[ YKNN GPJCPEG VJG GHHKECE[ QH IGPGVKE GPIKPGGTKPI HQT KORTQXGF CPF PQXGN RTQFWEV RCVJYC[U KP [GCUV DKQVGEJPQNQI[.
1 øWPEVKQPCN IGPGVKEU QH KPFWUVTKCN [GCUVUþ QH CPEKGPV UMKNNU CPF OQFGTP CRRNKECVKQPU
13
KP C UKPING [GCUV UVTCKP, VQ GPCDNG OCOOCNKCP J[FTQEQTVKUQPG RTQFWEVKQP HTQO C UKORNG ECTDQP UQWTEG NKMG INWEQUG. 6JG GPFQIGPQWU UVGTQN DKQU[PVJGVKE RCVJYC[ QH VJG [GCUV YCU TGTQWVGF VQ RTQFWEG EQORCVKDNG KPVGTOGFKCVGU, YJKEJ EQWNF UGTXG CU UWDUVTCVGU HQT VJG JGVGTQNQIQWU GP\[OGU KP VJG GPIKPGGTGF RCVJYC[. #NVJQWIJ [KGNFU OC[ UVKNN DG KORTQXGF, VJG HGCUKDKNKV[ QH VTCPUHGTTKPI EQORNGZ DKQU[PVJGVKE RCVJYC[U KPVQ [GCUV HQT RTQFWEVKQP RWTRQUGU JCU VJWU DGGP EQPENWUKXGN[ UJQYP.
1.ö üGTURGEVKXGU, TWNGU CPF TGIWNCVKQPU 6JKU DQQM FGUETKDGU KP ITGCV FGVCKN VJG CPCN[UKU, WVKNK\CVKQP, CPF CRRNKECVKQP QH IGPGVKE EJCTCEVGTKUVKEU QH KPFWUVTKCNN[ TGNGXCPV [GCUVU KP OQFGTP DKQVGEJPQNQI[. ôGPGVKE CPCN[UKU, TGEQODKPCPV &ó# VGEJPQNQI[, CPF PQXGN HWPEVKQPCN IGPQOKEU VGEJPQNQIKGU JCXG DGGP GORNQ[GF VQ FGHKPG VJG OQNGEWNCT DCUKU HQT KORQTVCPV DKQVGEJPQNQIKECN VTCKVU CPF RJGPQV[RGU. #NTGCF[, TGEQODKPCPV &ó# VGEJPQNQI[ CPF OGVCDQNKE GPIKPGGTKPI JCXG [KGNFGF PQXGN [GCUV UVTCKPU YKVJ KORTQXGF, QRVKOK\GF QT GXGP PGY RTQRGTVKGU CPF HWVWTG RQUUKDKNKVKGU UGGO XKTVWCNN[ WPNKOKVGF. %QPPGEVGF YKVJ VJGUG HWVWTG RGTURGEVKXGU QP DKQVGEJPQNQI[, VJG EWTTGPV NCEM QH RQNKVKECN CPF RWDNKE WPFGTUVCPFKPI CPF CEEGRVCPEG QH VJG WUG QH TGEQODKPCPV &ó# VGEJPQNQI[ QHHGTU UKPEGTG RTQDNGOU VQ HWTVJGT OKETQDKCN DKQVGEJPQNQIKECN FGXGNQROGPVU. 6Q D[RCUU CPF UQNXG EWTTGPV JGUKVCVKQPU CPF OKUEQPEGRVKQPU EQPEGTPKPI UCHGV[ QH TGEQODKPCPV VGEJPQNQI[ CPF VJG WUG QH ô/õ QTICPKUOU, EQPUWOGTU, UEKGPVKUVU, CPF KPFWUVT[ UJQWNF YQTM ENQUGN[ VQIGVJGT QP RTQRGT GFWECVKQP CPF EQOOWPKECVKQP RTQITCOU. õP VJG QVJGT JCPF, TGEQODKPCPV &ó# VGEJPQNQI[ CPF OQNGEWNCT IGPGVKEU CTG GUUGPVKCN KP DKQVGEJPQNQIKECN TGUGCTEJ CPF FGXGNQROGPV, HQT VJG KFGPVKHKECVKQP QH VCTIGVU HQT UVTCKP KORTQXGOGPV RTQITCOU VJTQWIJ ENCUUKECN (K.G. PQP-TGEQODKPCPV) IGPGVKEU UVTCVGIKGU. 6JKU KU FGRKEVGF ENGCTN[ KP 5VQMGU•U ÉòWCFTCPVÌ OQFGN HQT ENCUUKH[KPI TGUGCTEJ UVTCVGIKGU, YJKEJ KU DGCWVKHWNN[ KPVTQFWEGF CV VJG GPF QH %JCRVGT 4. #EEQTFKPI VQ VJG ÉúCUVGWT•U SWCFTCPVÌ OQFGN, YJCV KU PGGFGF KP OQFGTP DKQVGEJPQNQI[ KU C HWNN[ KPVGITCVGF CRRTQCEJ QH HWPFCOGPVCN OQNGEWNCT WPFGTUVCPFKPI QH OKETQDKCN RJ[UKQNQI[ CPF CP QRVKOCN RTCEVKECN WVKNK\CVKQP QH FGXGNQRKPI PGY RGTURGEVKXGU. õPN[ VJTQWIJ UWEJ CRRTQCEJGU YKNN YG DG CDNG VQ VCMG HWNN CFXCPVCIG QH VJG RJ[UKQNQIKECN RQVGPVKCN JCTDQWTGF KP VJG [GCUVU. úNC[KPI VJG ICOG D[ VJG TWNGU YKNN [KGNF QPN[ YKPPGTU.
#EMPQwNGFIGOGPVU øQT VJG EQORNGVKQP QH VJKU DQQM, O[ UKPEGTG VJCPMHWNPGUU IQGU VQ CNN VJG %JCRVGT CWVJQTU, YJQ JCXG FQPG C OCTXGNNQWU LQD KP EQORNGVKPI VJGKT GZVGPUKXG CPF WR-VQFCVG QXGTXKGYU. 9KVJQWV VJGKT GPVJWUKCUVKE CPF GZRGTV EQPVTKDWVKQPU, VJKU ÂRTQLGEV• YQWNF QDXKQWUN[ PQV JCXG DGGP RQUUKDNG. 5WDUGSWGPVN[, + YKUJ VQ VJCPM VJG 5GTKGU ©FKVQT 5VGHCP *QJOCPP HQT JKU WPNKOKVGF KPVGTGUV, GPVJWUKCUO, CPF -CDQXG CNN- JKU RCVKGPEG, YJKEJ UQOGVKOGU YCU SWKVG PGEGUUCT[. öCUV DWV PQV NGCUV, + YCPV VQ VJCPM
14
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O[ EQNNGCIWGU CV &5/, GURGEKCNN[ 6JQOCU úGVKV (CNUQ HQT JGNR YKVJ KNNWUVTCVKQPU), #PFTo 6GTYKUUEJC XCP 5EJGNVKPIC, 5GTIG &QPMGTU, óQrN XCP úGKL, /CTEQ XCP FGP $GTI, CPF ûQGN $QXGPDGTI, CPF CV &GNHV 6GEJPKECN ñPKXGTUKV[, GURGEKCNN[ ðCEM úTQPM, /CVV úKRGT, 5GH *GKLPGP CPF /KEMGN ðCPUGP, HQT OCP[ UVKOWNCVKPI CPF JGNRHWN FKUEWUUKQPU. %NGCTN[, VJKU YCU PQV VJG NCUV QH [GCUV KP DKQVGEJPQNQI[!
ÿGHGTGPEGU #NOGKFC /ð, úCKU %5 (1ÿÿü) %JCTCEVGTK\CVKQP QH VJG [GCUV RQRWNCVKQP HTQO VTCFKVKQPCN EQTP CPF T[G DTGCF FQWIJU. öGVV #RN /KETQDKQN 23:1ý4-1ý8 #TKUVKFQW #, úGPVKNNk / (2000) /GVCDQNKE GPIKPGGTKPI CRRNKECVKQPU VQ TGPGYCDNG TGUQWTEG WVKNK\CVKQP. %WTT õRKP $KQVGEJPQN 11:187-1ÿ8 #VVHKGNF ú÷ (1ÿÿ7) 5VTGUU VQNGTCPEG: VJG MG[ VQ GHHGEVKXG UVTCKPU QH KPFWUVTKCN DCMGT•U [GCUV. óCV $KQVGEJPQN 1ý:13ý1-13ý7 $CKNG[ ð© (1ÿÿ1) 6QYCTF C UEKGPEG QH OGVCDQNKE GPIKPGGTKPI. 5EKGPEG 2ý2:1üü8-1ü7ý $QNQVKP-øWMWJCTC /, 6QHHCPQ-óKQEJG %, #TVKIWGPCXG ø, &WEJCVGCW-óIW[GP ô, öGOCKTG /, /CTOGKUUG û, /QPVTQEJGT û, ûQDGTV %, 6GTOKGT /, 9KPEMGT ú, 9oUQNQYUMK-öQWXGN / (2000) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: 11. Kluyveromyces lactis. ø©$5 öGVV 487:üü-70 $QPGMCOR øð, õQUVGTQO ð (1ÿÿ4) õP VJG UCHGV[ QH Kluyveromyces lactis: C TGXKGY. #RRN /KETQDKQN $KQVGEJPQN 41:1-3 %JGP 5ö, %JKIGT / (1ÿ8ý) úTQFWEVKQP QH DCMGT•U [GCUV. +P: /QQ-[QWPI / (GF) %QORTGJGPUKXG $KQVGEJPQNQI[ ÷QN. 3. úGTICOQP úTGUU, õZHQTF, ñù, RR 42ÿ-4ýý %QTTCP *5 (1ÿ7ý) # JKUVQT[ QH DTGYKPI. &CXKU CPF %JCTNGU, óGYVQP #DDQVV, ñù &G &GMGP û* (1ÿüü) 6JG %TCDVTGG GHHGEV: C TGIWNCVQT[ U[UVGO KP [GCUV. ð ôGP /KETQDKQN 44:14ÿ-1ýü &GEQVVKIPKGU #, ôQHHGCW # (1ÿÿ7) %QORNGVG KPXGPVQT[ QH VJG [GCUV #$% RTQVGKPU. óCV ôGPGV 1ý:137-14ý &GOCKP #ö, 5QNQOQP ó# (1ÿ81) +PFWUVTKCN OKETQDKQNQI[. 5EK #OGTKECP 24ý, 42-ý1. &[GT ð/, %JCRKVCN &%, ùWCP ð9, /WNNGP û6, úGRRGTOCP #$ (2002) /GVCDQNKE GPIKPGGTKPI QH Saccharomyces cerevisiae HQT RTQFWEVKQP QH PQXGN NKRKF EQORQWPFU. #RRN /KETQDKQN $KQVGEJPQN ýÿ:224-230 øQWT[ ø (1ÿÿ7) *WOCP IGPGVKE FKUGCUGU: C ETQUU-VCNM DGVYGGP OCP CPF [GCUV. ôGPG 1ÿý:110 øTQP\C ô, +PIC #, /QPVK ú, 5EQVV ô, %CORQOGPQUK ú, /GPKEJKPK ú, õVVCIIKQ ö, ÷KCIIK 5, $WTPU ú#, ôQNF $, #DDQPFCPFQNQ # (2000) 6JG [GCUV Rý3 HWPEVKQPCN CUUC[: C PGY VQQN HQT OQNGEWNCT GRKFGOKQNQI[. *QRGU CPF HCEVU. /WVCVKQP ûGU 4ü2:2ÿ3-301 ôCTHKPMGN &ð, $CKNKU #/ (2002) óWENGQVKFG GZEKUKQP TGRCKT, IGPQOG UVCDKNKV[ CPF JWOCP FKUGCUG: PGY KPUKIJV HTQO OQFGN U[UVGOU. ð $KQOGF $KQVGEJPQN 2:ýý-ü0 ôQHHGCW # et al. (1ÿÿ7) 6JG ;GCUV ôGPQOG &KTGEVQT[. óCVWTG 387 (UWRRN):1-10ý *CTVYGNN ö (2002) óQDGN öGEVWTG. ;GCUV CPF ECPEGT. $KQUEK ûGR 22:373-3ÿ4 *QNNGPDGTI %ú, ôGNNKUUGP ô (1ÿÿ7) úTQFWEVKQP QH TGEQODKPCPV RTQVGKPU D[ OGVJ[NQVTQRJKE [GCUVU. %WTT õRKP $KQVGEJPQN 8:ýý4-ýü0 *WCPI ú, 5VTQHHGMQXC ù, %WRRQNGVVK ð, /CJCPV[ 5ù, 5ECTDQTQWIJ ô# (1ÿÿü) øWPEVKQPCN GZRTGUUKQP QH VJG E[UVKE HKDTQUKU VTCPUOGODTCPG EQPFWEVCPEG TGIWNCVQT KP [GCUV. $KQEJKO $KQRJ[U #EVC 1281:80-ÿ0
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ùKUGT ôö, ôGPV\UEJ /, ùNQUGT #ù, $CN\K ©, 9QNH &*, ôQHHGCW #, ûKQTFCP ðû (2001) ©ZRTGUUKQP CPF FGITCFCVKQP QH VJG E[UVKE HKDTQUKU VTCPUOGODTCPG EQPFWEVCPEG TGIWNCVQT KP Saccharomyces cerevisiae. #TEJ $KQEJGO $KQRJ[U 3ÿ0:1ÿý-20ý ùWTV\OCP %ú, øGNN ð9 (1ÿÿ8) 6JG ;GCUVU, C VCZQPQOKE UVWF[, 3TF ©FKVKQP. ©NUGXKGT 5EKGPEG úWDNKUJGTU, #OUVGTFCO öGCVJGTU 6 (2003) $KQEQPXGTUKQPU QH OCK\G TGUKFWGU VQ XCNWG-CFFGF EQRTQFWEVU WUKPI [GCUVNKMG HWPIK. ø©/5 ;GCUV ûGU 3:133-140 /CF\CM /, ôCKNNCTFKP %, $GEMGTKEJ ð/ (2003) *GVGTQNQIQWU RTQVGKP GZRTGUUKQP CPF UGETGVKQP KP VJG PQP-EQPXGPVKQPCN [GCUV ;CTTQYKC NKRQN[VKEC. ð $KQVGEJPQN, KP VJG RTGUU /CTVKP #%, &TWDKP &ô (2003) +ORCEV QH IGPQOG-YKFG HWPEVKQPCN CPCN[UGU QP EGNN DKQNQI[ TGUGCTEJ. %WTT õRKP %GNN $KQN 1ý, ü-13 óKGNUGP ð (2001) /GVCDQNKE GPIKPGGTKPI. #RRN /KETQDKQN $KQVGEJPQN ýý:2ü3-283 óWTUG ú (2002) %[ENKP FGRGPFGPV MKPCUGU CPF EGNN E[ENG EQPVTQN (PQDGN NGEVWTG). %JGODKQEJGO 3:ýÿü-ü03 úQYGNN %&, òWCKP &©, 5OCTV ù# (2003) 6JG KORCEV QH DTGYKPI [GCUV EGNN CIG QP HGTOGPVCVKQP RGTHQTOCPEG, CVVGPWCVKQP CPF HNQEEWNCVKQP. ø©/5 ;GCUV ûGU 3:14ÿ-1ý7 úTGVQTKWU +5 (2000) 6CKNQTKPI YKPG [GCUV HQT VJG PGY OKNNGPPKWO: PQXGN CRRTQCEJGU VQ VJG CPEKGPV CTV QH YKPG OCMKPI. ;GCUV 1ü:ü7ý-72ÿ òWG òò, 9KP\GNGT ©# (2002) öCTIG-UECNG OWVCIGPGUKU CPF HWPEVKQPCN IGPQOKEU KP [GCUV. øWPE +PVGIT ôGPQOKEU 2:1ÿ3-1ÿ8 ûQDKPUQP ð (1ÿÿ4) 6JG õZHQTF %QORCPKQP VQ 9KPG. õZHQTF ñKXGTUKV[ úTGUU, õZHQTF, ñù 5EJWNNGT &, %QTVG-ûGCN /, öGCQ % (2001) # FKHHGTGPVKCN OGFKWO HQT VJG GPWOGTCVKQP QH VJG URQKNCIG [GCUV Zygosaccharomyces bailii KP YKPG. ð øQQF úTQVGEV ü3:1ý70-1ý7ý 5OCTFQXC ð, úCXNQXC 5, ùQWNCMQXC * (2002) &GVGTOKPCVKQP QH QRVKOCN EQPFKVKQPU HQT CPCN[UKU QH Rý3 UVCVWU KP NGWMGOKE EGNNU WUKPI HWPEVKQPCN CPCN[UKU QH UGRCTCVGF CNNGNGU KP [GCUV. úCVJ õPEQN ûGU 8:24ý-2ý1 5YKPMGNU $9, XCP õQ[GP #ð, $QPGMCOR øð (1ÿÿ3) 6JG [GCUV Kluyveromyces lactis CU CP GHHKEKGPV JQUV HQT JGVGTQNQIQWU IGPG GZRTGUUKQP. #PVQPKG XCP öGGWYGPJQGM ü4:187-201 5\E\GDCTC ø/, %JCPFGNKGT %, ÷KNNGTGV %, /CUWTGN #, $QWTQV 5, &WRQTV %, $NCPEJCTF 5, ôTQKUKNNKGT #, 6GUVGV ©, %QUVCINKQNK ú, %CWGV ô, &GIT[UG ©, $CNDWGPC &, 9KPVGT ð, #EJUVGVVGT 6, 5RCIPQNK û, úQORQP &, &WOCU $ (2003) 6QVCN DKQU[PVJGUKU QH J[FTQEQTVKUQPG HTQO C UKORNG ECTDQP UQWTEG KP [GCUV. óCV $KQVGEJPQN 21:143-14ÿ 6TKXGFK ó$, ðCEQDUQP ôù, 6GUEJ 9 (1ÿ8ü) $CMGT•U [GCUV. %û% %TKV ûGX $KQVGEJPQN 4:7ý10ÿ ÷CP FGP $GTI ð#, ÷CP FGT öCMGP ùð, XCP õQ[GP #ð, ûGPPKGTU 6%, ûKGVXGNF ù, 5EJCCR #, $TCMG #ð, $KUJQR ûð, 5EJWNV\ ù, /Q[GT & (1ÿÿ0) Kluyveromyces CU C JQUV HQT JGVGTQNQIQWU IGPG GZRTGUUKQP: GZRTGUUKQP CPF UGETGVKQP QH RTQEJ[OQUKP. $KQVGEJPQNQI[ (ó;) 8:13ý-13ÿ ÷CP &KLMGP * (2001) %QPHGTGPEG ûGRQTV: 6JG 21UV +PVGTPCVKQPCN 5RGEKCNK\GF U[ORQUKWO QP ;GCUVU (+55;2001) QP Â$KQEJGOKUVT[, ôGPGVKEU, $KQVGEJPQNQI[ CPF ©EQNQI[ QH óQPEQPXGPVKQPCN ;GCUVU•. ø©/5 ;GCUV ûGU 2:42ÿ-432 ÷CWIJCP-/CTVKPK #, /CTVKPK # (1ÿÿý) øCEVU, O[VJU CPF NGIGPFU QP VJG RTKOG KPFWUVTKCN OKETQQTICPKUO. ð +PF /KETQDKQN 14:ý14-ý22 9CNMGT ô/ (1ÿÿ8) ;GCUV RJ[UKQNQI[ CPF DKQVGEJPQNQI[. ðQJP 9KNG[ CPF 5QPU, %JKEJGUVGT, ñù 9KPFGTKEMZ ð, *QNUDGGMU +, öCICVKG, õ, ôKQVU ø, 6JGXGNGKP ð, FG 9KPFG ð* (2002) øTQO HGCUV VQ HCOKPGþ CFCRVCVKQP VQ PWVTKGPV CXCKNCDKNKV[ KP [GCUV. +P: 6QRKEU KP %WTTGPV ôGPGVKEU, ÷QN. 1 (*QJOCPP 5, /CIGT 9*, GFU.), Â;GCUV UVTGUU TGURQPUGU•. 5RTKPIGT
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2 )GPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU 2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
#DUVTCEV ;GCUVU TGRTGUGPV JKIJN[ KORQTVCPV CPF XCNWCDNG QTICPKUOU HQT TGUGCTEJ CPF KPFWUVTKCN CRRNKECVKQPU. #NVJQWIJ Saccharomyces cerevisiae JCU JKUVQTKECNN[ DGGP C MG[ URGEKGUÿ QVJGT [GCUV IGPGTC CPF URGEKGU CTG KPETGCUKPIN[ KORQTVCPV. 6JG TCRKF GOGTIGPEG CPF GZRCPUKQP QH IGPQOKE CPF HWPEVKQPCN IGPGVKE KPHQTOCVKQP QP XCTKQWU [GCUVU KPFKECVGU C JKIJ NGXGN QH EQORNGZKV[ COQPIUV VJGUG QTICPKUOU. +V KU CRRCTGPV VJCV VJG S. cerevisiae IGPQOG YCU FWRNKECVGF UQOG 100 OKNNKQP [GCTU CIQ CPF VJCV UGXGTCN QH VJG QVJGT KORQTVCPV [GCUV URGEKGU FKXGTIGF RTKQT VQ VJKU FWRNKECVKQPÿ YJKNUV QVJGTU FKXGTIGF RQUV-RQN[RNQKFK\CVKQP QH VJG CPEGUVTCN [GCUV IGPQOG. *GTG YG RTGUGPV CP QXGTXKGY QH VJG XCTKQWU [GCUV IGPQOGU VJCV UGTXGU VQ JKIJNKIJV VJG EQORNGZKV[ QH [GCUVUÿ GURGEKCNN[ YKVJKP VJG EQPVGZV QH KPFWUVTKCNN[ TGNGXCPV UVTCKPU. 6JG EJCNNGPIGU HCEGF D[ KPFWUVTKCN [GCUV IGPGVKEKUVU CTG FKUEWUUGF CPF MG[ KUUWGU EQPEGTPKPI UVTCVGIKGU QH OCVKPIÿ OWVCIGPGUKUÿ URJGTQRNCUV HWUKQP CPF E[VQFWEVKQP CTG RTGUGPVGF. ©ZCORNGU QH UVTCKP KORTQXGOGPVU CEJKGXGF XKC ENCUUKECN IGPGVKEU CTG RTGUGPVGFÿ CNQPI YKVJ CP CPCN[UKU QH OQNGEWNCT IGPGVKEU QH OCNVQUG OGVCDQNKUO KP KPFWUVTKCN [GCUVUÿ YJKEJ UGTXGU VQ JKIJNKIJV VJG WUGHWNPGUU QH EQODKPGF OQNGEWNCT CPF ENCUUKECN IGPGVKEU KP UVTCKP KORTQXGOGPV.
2.1 +PVTQFWEVKQP 8CTKQWU [GCUVU CTG WUGF HQT VGEJPQNQIKECN RTQEGUUGU VJCV TCPIG HTQO VJG CPEKGPV CTVU QH DTGCFÿ YKPG CPF DGGT OCMKPIÿ VQ VJG OQFGTP CRRNKECVKQP QH JGVGTQNQIQWU RTQVGKP RTQFWEVKQP (UGG QVJGT EJCRVGTU KP VJKU XQNWOGþ 5RGPEGT CPF 5RGPEGT 1990þ 4GGF CPF ýCIQFCYKVJCPC 1991). /QFGTP [GCUV VGEJPQNQI[ TGRTGUGPVU C XCUV KPFWUVTKCN UGEVQT YQTVJ CDQWV 75üû0 DKNNKQP RGT CPPWO. %QPUWOGTU QH [GCUVU CPF [GCUVDCUGF RTQFWEVU FGOCPF EQPVKPWCNN[ KORTQXGF SWCNKV[ CPF GEQPQOKEU. 6JGTGHQTGÿ [GCUV VGEJPQNQIKUVU CTG WPFGT ITQYKPI RTGUUWTG VQ GPJCPEG VJG RGTHQTOCPEGU QH KPFWUVTKCN UVTCKPU KP EQOOGTEKCN RTQEGUUGU. #P KP FGRVJ TGXKGY QH KPFWUVTKCNN[ TGNGXCPV VTCKVU KU DG[QPF VJG UEQRG QH VJKU CTVKENGÿ DWV DTKGHN[ÿ jWUV UQOG QH VJG MG[ RTQRGTVKGU VJCV [GCUV KPFWUVTKGU NQQM VQ KORTQXG KPENWFG [KGNF QH [GCUV DKQOCUU QP EJGCR UWDUVTCVGUÿ HNQY QH UWICTU VJTQWIJ VQ GVJCPQN CPF %úù RTQFWEVKQPÿ TGURKTCVQT[ OGVCDQNKUO YJGTG OCZKOWO EQPXGTUKQP QH ECTDQP UQWTEG VQ EGNN OCUU KU TGSWKTGFÿ NGXGNU QH GP\[OGU CPF OGVCDQNKE KPVGTOGFKCVGU HQT RTQFWEKPI 4&ø EJGOKECNU QT FTWI RTGEWTUQTUÿ RTQVGKP CPF COKPQ CEKF EQORNGOGPVU HQT [GCUV GZVTCEVU Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeats Springer-Verlag Berlin Heidelberg 2003
1÷
2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
CPF HGGF [GCUVUÿ PWENGQVKFG EQPVGPVU HQT HNCXQWT GPJCPEGTUÿ XKVCOKP RTQFWEVKQPÿ CPF RTQFWEVKXKV[ (CPF UGETGVKQP) QH HQTGKIP RTQVGKPU. # OCjQT TGSWKTGOGPV HQT UVTCKP KORTQXGOGPV KU KP VJG CTGC QH VQNGTCPEG CPF CFCRVCDKNKV[ QH EGNNU VQ OKETQCPF OCETQ-GPXKTQPOGPVCN UVTGUUGU KORCTVGF D[ KPFWUVTKCN CRRNKECVKQPU. ;GCUVUÿ NKMG CNN EGNNWNCT QTICPKUOUÿ PGGF VQ OCKPVCKP KPVGTPCN EQPFKVKQPU YKVJKP C FGHKPGF TCPIG KP QTFGT VQ ITQY CPF HWPEVKQP QRVKOCNN[. +V UJQWNF DG TGOGODGTGF VJCV KP IGPGTCNÿ [GCUVU JCXG GXQNXGF VQ QEEWR[ GEQNQIKECN PKEJGU VJCV CTG SWKVG FKHHGTGPV HTQO VJQUG GPEQWPVGTGF KP NCDQTCVQTKGU QT KP KPFWUVTKCN RTQEGUUGU. 6JG HCEV KU VJCV [GCUVU TCTGN[ GPEQWPVGT KFGCN RJ[UKQNQIKECN EQPFKVKQPU FWTKPI VJGKT KPFWUVTKCN NKHG URCPU. 6JGTGHQTGÿ VJG CDKNKV[ QH C [GCUV UVTCKP VQ CFCRV VQ XCTKQWU EJCNNGPIGUÿ YJKEJ HCNN QWVUKFG QH VJG QRVKOCN GPXKTQPOGPVCN EQPFKVKQPU HQT ITQYVJÿ YKNN IQXGTP KVU WUGHWNPGUU CPF CRRNKECDKNKV[ VQ KPFWUVTKCN CRRNKECVKQPU. 6JGUG EQPUKFGTCVKQPU QH VCTIGVU HQT UVTCKP KORTQXGOGPV CTG UQOGYJCV EWTUQT[ÿ PGXGTVJGNGUU VJG[ JKIJNKIJV VJG FKXGTUG TCPIG QH HGCVWTGU VJCV [GCUV VGEJPQNQIKUVU TGSWKTG HTQO UVTCKPU. úH EQWTUG VJG DKQNQIKECN DCUGU QH VJGUG HGCVWTGU CTG EQORNGZ CPF CU UWEJÿ KORTQXKPI VJGUG CVVTKDWVGU KP UVTCKPU KU C EJCNNGPIKPI VCUM. +ORTQXGOGPV QH [GCUV IGPGVKEU ECP DG CEJKGXGF VJTQWIJ ENCUUKECNÿ QT OQNGEWNCT OCPKRWNCVKQPU (TGEQODKPCPV (T)øý# QT IGPGVKE GPIKPGGTKPI). 5Q-ECNNGF ENCUUKECN IGPGVKEU TGHGTU VQ VGEJPKSWGU QH DTGGFKPI (CNUQ TGHGTTGF VQ CU OCVKPI QT J[DTKFK\CVKQP)ÿ OWVCIGPGUKU CPF RTQVQRNCUVöURJGTQRNCUV HWUKQP. ©CEJ QH VJGUG JCU KVU CFXCPVCIGU CPF FKUCFXCPVCIGUÿ YJKEJ CTG FKUEWUUGF KP 5GEVKQP 3. 6JG EJCNNGPIG QH KORTQXKPI KPFWUVTKCN [GCUVU KU OCFG GURGEKCNN[ FKHHKEWNV D[ VJG IGPGVKE PCVWTG QH VJGUG [GCUVU. 7PNKMG NCDQTCVQT[ UVTCKPU QH Saccharomyces cerevisiae VJCV JCXG DGGP DTGF VQ DG IGPGVKECNN[ VTCEVCDNGÿ KPFWUVTKCN [GCUV UVTCKPU CTGÿ IGPGTCNN[ÿ OWEJ NGUU COGPCDNG VQ IGPGVKE OCPKRWNCVKQP. 6JWUÿ YJGTGCU NCDQTCVQT[ UVTCKPU CTG WUWCNN[ JCRNQKFU YKVJ UVCDNG OCVKPI V[RGUÿ QT FKRNQKFU VJCV ECP DG URQTWNCVGF VQ IKXG IGPG UGITGICVKQPU VJCV QDG[ /GPFGN'U NCYUÿ KPFWUVTKCNN[ TGNGXCPV [GCUV UVTCKPU CTG QHVGP RQN[RNQKF QT CPGWRNQKF CPF TCTGN[ GZJKDKV /GPFGNKCP UGITGICVKQP. #NVJQWIJ DTGGFKPI UVTCVGIKGU ECP DG CRRNKGF VQ [GCUVU YKVJ EQORNGZ IGPQOGUÿ XGT[ JKIJ PWODGTU (> 10õ) QH J[DTKFU PGGF VQ DG UETGGPGF KP QTFGT VQ QDVCKP FGUKTGF IGPQV[RGU. +P UQOG ECUGUÿ KPFWUVTKCN UVTCKPU CTG CUGZWCNÿ QT CURQTQIGPQWU CPF VJGTGHQTG DTGGFKPI UVTCVGIKGU ECPPQV DG GORNQ[GF VQ KORTQXG IGPGVKE DCEMITQWPFÿ KP YJKEJ ECUG OWVCIGPGUKU KU CP CNVGTPCVKXG UVTCVGI[. #NUQÿ UQOGVKOGU KV KU FGUKTCDNG VQ EQODKPG VTCKVU QH [GCUVU HTQO FKHHGTGPV IGPGTC QT URGEKGUÿ KP YJKEJ ECUG EGNN HWUKQP ECP DG GORNQ[GF. #P CFFGF EQORNGZKV[ KP IGPGVKE OCPKRWNCVKQP QH [GCUVU KU VJG OWNVKIGPKE PCVWTG QH OQUV QH VJG VTCKVU VJCV YG UGGM VQ KORTQXG. 5KPING IGPG EJCPIGU TCTGN[ JCXG CP GHHGEV QP TGNGXCPV RJGPQV[RGU CPF UQ OCjQT KORTQXGOGPVU XKC ENCUUKECN IGPGVKE QT Tøý# CRRTQCEJGU JCXG VQ DG HQWPFGF QP C IQQF WPFGTUVCPFKPI QH VJG EGNNWNCT CPF OQNGEWNCT DKQNQI[ IQXGTPKPI VCTIGVGF VTCKVUÿ UQ VJCV GKVJGT UWKVCDNG UGNGEVKQP CPF GPTKEJOGPV EQPFKVKQPU ECP DG FGUKIPGF HQT KUQNCVKPI ENCUUKECNN[ KORTQXGF EGNN V[RGUÿ QT VJG TGNGXCPV IGPGU ECP DG VCTIGVGF KP Tøý# UVTCVGIKGU. +P VJKU EJCRVGT YG EQORCTG CPF EQPVTCUV VJG IGPQOKEU QH KPFWUVTKCNN[ TGNGXCPV [GCUVUÿ CPF TGXKGY VJG UVTCVGIKGU HQT KORTQXGOGPV QH UVTCKPU KP NKIJV QH VJGKT IGPGVKE EQORNGZKVKGU.
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
19
2.2 )GPGVKE RTQRGTVKGU QH KPFWUVTKCN [GCUVU 6JG OCjQT KPFWUVTKCNN[ WUGHWN [GCUVU KPENWFG UVTCKPU QH S. cerevisiaeÿ QVJGT Saccharomyces UR.ÿ CPF URGEKGU QH Candidaÿ Hansenula (Pichia)ÿ Hanseniaspora (Kloeckera)ÿ Kluyveromycesÿ Phaffiaÿ Rhodotorulaÿ Torulasporaÿ Torulopsisÿ Yarrowiaÿ CPF Zygosaccharomyces (5RGPEGT CPF 5RGPEGT 1990þ 4GGF CPF ýCIQFCYKVJCPC 1991). 6CDNG 1 NKUVU UQOG QH VJG KPFWUVTKCN WUGU QH TGRTGUGPVCVKXG [GCUVU CPF KPFKECVGU VJG FKXGTUKV[ QH CRRNKECVKQPU. #U YGNN CU DGKPI CP GZVTGOGN[ KORQTVCPV GEQPQOKE [GCUVÿ S. cerevisiae KU VJG OQUV UVWFKGF [GCUV JCXKPI DGGP CV VJG XCPIWCTF QH HWPFCOGPVCN CPF CRRNKGF DKQNQI[ TGUGCTEJ HQT OCP[ FGECFGU. *QYGXGTÿ VJG FGGR CPF TCRKFN[ ITQYKPI MPQYNGFIG VJCV YG JCXG QH S. cerevisiae KU DCUGF NCTIGN[ WRQP UVWFKGU YKVJ NCDQTCVQT[ OWVCPVU. 6JG TGNGXCPEG QH VJGUG UVWFKGU VQ KPFWUVTKCN UVTCKPU QH S. cerevisiae KU UQOGYJCV SWGUVKQPCDNG UKPEG KP OCP[ ECUGU VJG IGPQOKEU QH UVTCKPU WUGF KP KPFWUVT[ KU UKIPKHKECPVN[ FKHHGTGPV HTQO VJCV QH V[RKECN NCDQTCVQT[ UVTCKPU. óJGP YG EQPUKFGT VJG PQP-Saccharomyces URGEKGU QH KPFWUVTKCNN[ TGNGXCPV [GCUVUÿ VJG CXCKNCDNG KPHQTOCVKQP KU GXGP NGUU GZVGPUKXG. ýGXGTVJGNGUUÿ KORQTVCPV KPHQTOCVKQP KU CTKUKPI HTQO IGPQOKE GZRNQTCVKQPU QH VJGUG [GCUVU. 2.2.1 #P QXGTXKGY QH VJG IGPQOG CPF HWPEVKQPCN IGPGVKE CPCN[UKU QH NCDQTCVQT[ S. cerevisiae UVTCKPU: C DCUKU HQT EQORCTKUQP YKVJ KPFWUVTKCN [GCUVU 7PFGTUVCPFKPI JQY VJG IGPQOG QH C [GCUV UVTCKP EQPHGTU VJG EJCTCEVGTKUVKEU GUUGPVKCN HQT GHHKEKGPV KPFWUVTKCN RGTHQTOCPEG KU ETWEKCN VQ FGXGNQRKPI VCTIGVU CPF UVTCVGIKGU HQT UVTCKP KORTQXGOGPV. 6JG OQUV RTQOKUKPI CRRTQCEJ VQ WPTCXGNNKPI VJG TGNC 6CDNG 1. #RRNKECVKQPU QH UQOG TGRTGUGPVCVKXG KPFWUVTKCN [GCUVU )GPWU Saccharomyces
5RGEKGU cerevisiae
Saccharomyces Candida Candida Candida Rhodotorula Pichia Pichia
various sp utilis paraffinica lipolytica sp. angusta sp.
Kluyveromyces Kluyveromyces Kluyveromyces Hansenula Torulopsis Torulopsis Zygosaccharomyces
fragilis marxianus lactis sp. sp. sp. rouxii
#RRNKECVKQP $CMKPIÿ DTGYKPIÿ YKPGÿ FKUVKNNKPIÿ HWGN CNEQJQNÿ CPF HQTGKIP RTQVGKPUÿ KPXGTVCUGÿ GTIQUVGTQN $TGYKPIÿ YKPGÿ UCMGÿ CO[NCUGÿ OGNKDKCUG òQQF [GCUVÿ PWENGQVKFGU òQQF [GCUV %KVTKE CEKF %CTQVGPGÿ KVCEQPKE CEKF òQTGKIP RTQVGKPU òQQF [GCUVÿ UVGTQKF RTGEWTUQTUÿ ½-ICNCEVQUKFCUGÿ TKDQHNCXKP +PXGTVCUGÿ HQQF [GCUV %JGGUG òQQF [GCUVÿ ¾-ICNCEVQUKFCUG 5VGTQKF RTGEWTUQTU òQQF [GCUV ôNWEQPKE CEKFÿ HQQF [GCUV 5Q[ UCWEGÿ OKUQ
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
VKQPUJKR DGVYGGP VJG [GCUV IGPQOG CPF EGNN HWPEVKQPU KPXQNXGU CP KPVGITCVGF CPCN[VKECN CRRTQCEJ. òKIWTG 1 UJQYU VJG KPVGITCVGF CPCN[VKECN RTQEGUUÿ YJKEJ KPXQNXGU CURGEVU QH ENCUUKECN IGPGVKEUÿ OQNGEWNCT DKQNQI[ÿ DKQEJGOKUVT[ÿ EGNN DKQNQI[ CPF EQORWVCVKQPCN VGEJPKSWGU. /CP[ QH VJG OGVJQFU WUGF KP VJG CPCN[UGU EQPVKPWG VQ DG FGXGNQRGF CPF TGHKPGFÿ CPF FCVC JCPFNKPI (DKQKPHQTOCVKEU) KUUWGU TGOCKP C RCTVKEWNCT EJCNNGPIG. *QYGXGTÿ YG HQTGUGG VJCV VJG KPVGITCVGF CRRTQCEJ YKNN DG KPETGCUKPIN[ KORQTVCPV VQ FGHKPKPI ETKVKECN FGVGTOKPCPVU QH KPFWUVTKCN RGTHQTOCPEG D[ [GCUV UVTCKPU. 4GIWNCTN[ WRFCVGF KPHQTOCVKQP EQPEGTPKPI VJG IGPQOKE CPF HWPEVKQPCN CPCN[UKU QH [GCUVU KU CXCKNCDNG QP C PWODGT QH GZVGPUKXG FCVCDCUGU. 6JGUG KPENWFG VJG ôoPQNGXWTGU RTQjGEV YGD UKVGU (UGG 5QWEKGV GV CN. ù000)ÿ VJG 5VCPHQTF ôGPQOG øCVCDCUG (5ôø)ÿ VJG /WPKEJ +PHQTOCVKQP %GPVGT HQT 2TQVGKP 5GSWGPEGU %QORTGJGPUKXG ;GCUV ôGPQOG øCVCDCUG (/+25 %;ôø)ÿ CPF VJG ;GCUV 2TQVGQOG øCVCDCUG (;2ø). 5GG ôQHHGCW (ù000)ÿ 4QUU-/CEFQPCNF (ù000) CPF 5QWEKGV GV CN. (ù000) HQT C DTQCFGT NKUV QH WUGHWN YGD UKVGU. #U CP GZCORNG QH JQY VJG KPVGITCVGF CRRTQCEJ VQ [GCUV CPCN[UKU JCU DGGP CRRNKGF D[ TGUGCTEJ ITQWRU CTQWPF VJG YQTNFÿ YG RTQXKFG C DTKGH QXGTXKGY QH NCDQTCVQT[ S. cerevisiae IGPQOKEU. 6JKU QXGTXKGY CNUQ UGTXGU CU C RQKPV QH TGHGTGPEG HQT QWT NCVGT EQORCTKUQP YKVJ KPFWUVTKCNN[ TGNGXCPV [GCUV UVTCKPU. 6JG PWENGCT IGPQOG QH VJG UGSWGPEGF S. cerevisiae UVTCKP 5ù÷÷E VQVCNU 1ù /D. 6JG UVTCKP ECTTKGU 1õ EJTQOQUQOGU XCT[KPI HTQO ù30MD VQ 1.ñ /D KP NGPIVJ. 6JGTG CTG OCP[ FWRNKECVGF EJTQOQUQOCN TGIKQPU KP S. cerevisiae UWIIGUVKPI C YJQNG IGPQOG FWRNKECVKQP CRRTQZKOCVGN[ 10÷ [GCTU CIQ (óQNHG CPF 5JKGNFU 199ûþ 5GQKIJG CPF óQNHG 199÷). ôGPG ENWUVGT CPCN[UKU UWIIGUVU CDQWV ññ OCjQT TGIKQPU QH ENWUVGTGF IGPG FWRNKECVKQPUÿ PQV KPENWFKPI VGNQOGTKE CPF UWDVGNQOGTKE TGIKQPUÿ YJKEJ CTG YGNN MPQYP HQT JKIJ NGXGN UKOKNCTKV[ (/GYGU GV CN. 199ûþ óQNHG CPF 5JKGNFU 199ûþ 5GQKIJG CPF óQNHG 199÷). %QODKPKPI IGPG QTFGT YKVJ IGPG FWRNKECVKQP FCVC CUUKIPU OQTG VJCP ÷0ð QH VJG IGPQOG KPVQ 11ù RCKTGF TGIKQPUÿ YKVJ VJG NCTIGUV WPRCKTGF ICR DGKPI QPN[ 3õ IGPGU NQPI (óQPI GV CN. ù00ù). 5GXGTCN QH VJG UKUVGT TGIKQPU URCP EGPVTQOGTGUÿ YJKEJ CNNQYU EGPVTQOGTG RCKTU VQ DG KPFKECVGF CU HQNNQYU: +ö8++ÿ ++ö+8ÿ +++öï+8ÿ 8ö+ïÿ ïöï++ÿ CPF ï+++öï8. LKMGN[ EGPVTQOGTG RCKTKPI CNUQ CRRGCTU DGVYGGP EJTQOQUQOGU 8+öï8+ CPF 8+++öï+ÿ CNVJQWIJ KP VJGUG ECUGU VJG UKUVGT TGIKQPU GPF ENQUG VQ EGPVTQOGTGU TCVJGT VJCP URCPPKPI VJGO. /CP[ QH VJG OCjQT FWRNKECVKQPU CTG KP VJG UCOG QTKGPVCVKQP TGNCVKXG VQ VJG EGPVTQOGTG KP VJG VYQ EQRKGUÿ UWIIGUVKPI VJCV TGEKRTQECN VTCPUNQECVKQP JCU DGGP VJG OCKP HQTO QH NCTIG-UECNG TGCTTCPIGOGPV YKVJKP VJG [GCUV IGPQOG (5GQKIJG CPF óQNHG 199÷). 6JG S. cerevisiae IGPQOG YCU OCRRGF RTKQT VQ IGPQOG UGSWGPEKPI WUKPI OGVJQFU UWEJ CU ENCUUKECN VGVTCF CPCN[UKUÿ TCPFQO URQTG CPCN[UGUÿ VTKUQOKE CPCN[UKUÿ OKVQVKE ETQUUKPI-QXGTÿ CPF EJTQOQUQOG NQUU CPF VTCPUHGT (/QTVKOGT CPF 5EJKNF 19÷1). 6JG CEEWOWNCVGF OCRRKPI FCVC RTQXKFGF KPHQTOCVKQP QP EJTQOQUQOCN NQECVKQPU CPF QTFGTU QH OCP[ QH VJG IGPGU YKVJ TGURGEV VQ EGPVTQOGTGU. 6JG FGTKXGF OCRU YGTG RWDNKUJGF KP C UGTKGU QH TGIWNCT WRFCVGU D[ 4.î. /QTVKOGT CPF EQNNGCIWGU (EKVGF KP %JGTT[ GV CN. 199û). 6JG UGSWGPEKPI QH UVTCKP 5ù÷÷E RTQXKFGF HQT C RJ[UKECN OCR QH VJG IGPQOG (%JGTT[ GV CN. 199û)ÿ YJKEJ KU CXCKNCDNG HTQO VJG 5VCPHQTF 7PKXGTUKV['U Saccharomyces ôGPQOG øCVCDCUG. 5VTCKP 5ù÷÷E ECTTKGU CDQWV õ000 IGPGU QT QRGP TGCFKPI HTCOGU (ú4òU) (øWjQP 199õ). 6JG GZCEV PWODGT
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
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)GPQOG %NCUUKECN IGPGVKE OCRRKPIÿ øý# UGSWGPEKPIÿ KFGPVKHKECVKQP QH QRGP TGCFKPI HTCOGUÿ IGPG QTFGTKPI
)GPG RTQFWEV CPF HWPEVKQP òWPEVKQPCN CPCN[UGUÿ KP UKNKEQ (FCVCDCUG TGHGTGPEG)ÿ FGNGVKQPU (MPQEM-QWVU)ÿ VTCPURQUQP KPUGTVKQPUÿ VYQJ[DTKFU GVE
2TQVGQOG ù-ø IGNUÿ RTQVGKP UGSWGPEKPI
6TCPUETKRVQOG
/GVCDQNQOG
/KETQCTTC[U CPF QVJGT INQDCN GZRTGUUKQP CPCN[UGU
#PCN[UGU QH KPVGTOGFKCVGUÿ EQHCEVQTUÿ RTQFWEVU QH RCVJYC[U GVE.
$KQKPHQTOCVKEU CPF EQORWVCVKQPCN OQFGNNKPI
)GPG HCOKNKGUÿ GZRTGUUKQP ENWUVGTU, RTQVGKP EQORNGOGPV, EQ-QTFKPCVGF TGURQPUGU CPF RCVJYC[ CEVKXKVKGU
(WPEVKQPKPI %GNN
(KI. 1. #P KPVGITCVGF CPCN[VKECN CRRTQCEJ VJCV NKPMU VJG IGPQOG VQ EGNN HWPEVKQP. # [GCUV UVTCKP ECP DG CPCN[\GF XKC IGPQOKEU VQ RTQXKFG C FGVCKNGF IGPQOG UGSWGPEG CPF RJ[UKECN OCR QH EJTQOQUQOGU. 8CTKQWU VGEJPKSWGU QH HWPEVKQPCN CPCN[UGU CTG WUGF VQ FGVGTOKPG IGPG RTQFWEVUÿ CPF IGPG HWPEVKQPUÿ CU YGNN CU INQDCN IGPG GZRTGUUKQP (VTCPUETKRVQOKEU) CPF INQDCN RTQVGKP (RTQVGQOKEU) EQORNGOGPVU KP EGNNU WPFGT URGEKHKE EQPFKVKQPU. 6JG VTCPUETKRVQOG CPF RTQVGQOG FCVC KU EQWRNGF YKVJ EQORWVCVKQPCN CPCN[UGU VQ KFGPVKH[ IGPG HCOKNKGU CPF ENWUVGTUÿ CPF EQ-QTFKPCVGF TGURQPUGU D[ EGNNU KPENWFKPI EQPVTQN QH RCVJYC[U. #PCN[UKU QH VJG OGVCDQNKVGU RTGUGPV KP EGNNU WPFGT VJG UCOG EQPFKVKQPU (OGVCDQNQOKEU) RTQXKFGU KPHQTOCVKQP QP HNWZ QH RCVJYC[U CPF GP\[OG CEVKXKVKGU. 6JKU KPVGITCVGF CRRTQCEJ CNNQYU HQT KFGPVKHKECVKQP QH VJG DTQCF IGPG HWPEVKQPU VJCV CTG TGSWKTGF VQ RTQXKFG VJG KORQTVCPV EJCTCEVGTKUVKEU QH [GCUV EGNNU WPFGT FKHHGTGPV EQPFKVKQPU.
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
QH EQFKPI ú4òU TGOCKPU WPEGTVCKP ($CUTCK GV CN. 199ûþ îQYCNE\WM GV CN. 1999þ ôQHHGCW ù000þ 4QUU-/CEFQPCNF ù000). 5VTCKP 5ù÷÷E KU VJG OQUV EQOOQPN[ WUGF IGPGVKE DCEMITQWPF KP TGUGCTEJ NCDQTCVQTKGU CPF YCU FGTKXGF HTQO C PCVWTCN FKRNQKF KUQNCVGÿ ©/93 KUQNCVGF D[ ©OKN /TCM HTQO C TQVVKPI HKI KP %CNKHQTPKC KP 193÷ (/QTVKOGT CPF ,QJPUVQP 19÷õ). 6JG TGNCVKQPUJKR DGVYGGP VJG UGSWGPEGF S. cerevisiae UVTCKP CPF KPFWUVTKCN UVTCKPU KU WPENGCTÿ CNVJQWIJ KV JCU DGGP GUVKOCVGF VJCV ÷ñð QH VJG 5ù÷÷E IGPQOG EQOGU HTQO ©/93ÿ YJKEJ YCU RTQDCDN[ C YKPG [GCUV (/QTVKOGT ù000). 5KIPKHKECPVN[ÿ YKVJ TGICTFU VQ DCMKPIÿ FKUVKNNKPI QT DTGYKPI CRRNKECVKQPUÿ UVTCKP 5ù÷÷E KU KPECRCDNG QH WVKNK\KPI OCNVQUGÿ VJG OCjQT UWICT CXCKNCDNG VQ [GCUVU KP RNCKP FQWIJUÿ FKUVKNNGT'U OCUJGU QT DTGYGT'U YQTVÿ GXGP VJQWIJ VJG [GCUV IGPQOG UGSWGPEKPI RTQjGEV JCU KFGPVKHKGF VYQ EQRKGU QH VJG MAL NQEK (òGWGTOCPP GV CN. 199ñ). 5VTCKP 5ù÷÷E KU KPECRCDNG QH HGTOGPVKPI OCNVQUG DGECWUG KV NCEMU C HWPEVKQPCN MAL TGIWNCVQT[ RTQVGKP (8. *KIIKPUÿ RGTU. EQOO.). ýGXGTVJGNGUUÿ KPHQTOCVKQP QDVCKPGF HTQO UVWFKGU QH UVTCKP 5ù÷÷E CPF QVJGT NCDQTCVQT[ UVTCKPU FQGU RTQXKFG C RNCVHQTO HQT WPFGTUVCPFKPI KPFWUVTKCN [GCUV UVTCKPU. 6JG HWPFCOGPVCN EJCNNGPIG HQT [GCUV DKQNQIKUVU KU VQ FGVGTOKPG YJCV VJG HWPEVKQPU QH VJG õ000 QT UQ S. cerevisiae IGPGU CTG. #V VJG VKOG QH VJG RWDNKECVKQP QH VJG IGPQOG UGSWGPEGÿ QPN[ CDQWV 30ð QH VJG ú4òU EQTTGURQPFGF VQ IGPGU YJQUG RTQFWEV HWPEVKQPU YGTG EJCTCEVGTK\GF D[ EQPXGPVKQPCN OGCPUÿ KPENWFKPI HWPEVKQPCN CUUC[. #DQWV QPG JCNH QH VJG TGOCKPKPI ú4òU UJQYGF UQOG FGITGG QH JQOQNQI[ QT EQOOQP OQVKHU YKVJ IGPGU QH [GCUV QT QVJGT QTICPKUOU YJQUG RTQFWEVU CTG HWPEVKQPCNN[ EJCTCEVGTK\GFÿ CPF CU UWEJ C TQNG KP [GCUV ECP DG KPHGTTGF. 6JG TGOCKPKPI ù000 QT UQ ú4òU JCF PQ MPQYP HWPEVKQP. 6JG /+25 %QORTGJGPUKXG ;GCUV ôGPQOG øCVCDCUG NKUVU ÷û÷ IGPG RTQFWEVU CU DGKPI GUUGPVKCN HQT XKCDKNKV[ÿ 31ñ÷ CU DGKPI FKURGPUCDNG HQT XKCDKNKV[ÿ YJKNUV í÷ IGPGU YGTG HQWPF VQ DG CODKIWQWU YKVJ TGURGEV VQ VJGKT GHHGEVU QP XKCDKNKV[. 8CTKQWU CWVJQTU JCXG CNTGCF[ FGUETKDGF MG[ GNGOGPVU QH VJG KPVGITCVGF CRRTQCEJ VQ GNWEKFCVKPI VJG HWPEVKQPU QH VJG PQXGN IGPGU (óKP\GNGT CPF øCXKU 199ûþ úNKXGT GV CN. 199÷þ ôQHHGCW ù000þ 4QUU-/CEFQPCNF ù000þ úNKXGT ù00ù). #RRTQCEJGU VQ IGPG HWPEVKQPCN CPCN[UKU JCXG KPENWFGF KP UKNKEQ EQORCTKUQPU DGVYGGP GZKUVKPI CPF FGXGNQRKPI FCVCDCUGUÿ EQPUVTWEVKQP QH FGNGVKQP-OWVCPV NKDTCTKGU CPF VTCPURQUQP KPUGTVKQP NKDTCTKGUÿ øý# (OKETQ)CTTC[Uÿ VYQ-J[DTKF CPCN[UKU CPF RTQVGQOKEU. $KCPEJK GV CN. (ù001) ECTTKGF QWV CPCN[UKU QH UVTCKPU FGNGVGF HQT ñõí QH VJG RQQTN[ FGHKPGF IGPGU. #DQWV 30ð QH VJG FGNGVCPVU GZJKDKVGF RJGPQV[RGU CUUQEKCVGF YKVJ TGURQPUG VQ KPJKDKVQTUö UVTGUUGU CPF OCP[ YGTG RNGKQVTQRKE. +V YCU UWIIGUVGF VJCV UQOG IGPGU YKVJ WPMPQYP HWPEVKQPU OCMG EQPVTKDWVKQPU VQ EGNN HWPEVKQPU VJCV CTG KFGPVKECN VQ IGPGU YJQUG HWPEVKQPU CTG CNTGCF[ FGHKPGF. 6JG XCUV OCjQTKV[ QH IGPG FGNGVKQPU ECWUG JKIJN[ UKIPKHKECPV CNVGTCVKQPU KP VJG GZRTGUUKQP QH CV NGCUV QPG QVJGT IGPG CU TGXGCNGF D[ YJQNG-IGPQOG CTTC[U (*WIJGU GV CN. ù000). 6JKU OGCPU VJCV OWVCVKQPU ECP JCXG OCP[ RQUUKDNG UGEQPFCT[ GHHGEVU DG[QPF FGNGVKPI VJG RTKOCT[ HWPEVKQP QH C IGPG'U RTQFWEVÿ YJKEJ CFFU CPQVJGT NGXGN QH EQORNGZKV[ VQ HWPEVKQPCN CPCN[UKU. òGCVJGTUVQPG CPF $TQCFKG (ù00ù) JCXG WUGF EQORWVCVKQPCN CPCN[UKU VQ VT[ CPF QXGTEQOG VJKU RTQDNGOÿ CPF CTIWG VJCV IGPG GZRTGUUKQP HQTOU C 'UECNG-HTGG' PGVYQTM UKOKNCT VQ CTVKHKEKCN PGVYQTMU UWEJ CU RQYGT ITKFU CPF VJG KPVGTPGV. 6JGKT OQFGN RTGFKEVU VJCV VJG IGPG PGVYQTM UVTWEVWTG KU QTICPK\GF HQT TQDWUVPGUU CPF
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JGNRU OCMG QTICPKUOU TGUKUVCPV VQ FGNGVGTKQWU GHHGEVU QH OWVCVKQPUÿ VJWU EQPHGTTKPI GXQNWVKQPCT[ CFXCPVCIGU. /KETQCTTC[ RTQXKFGU FCVC QP OCUU-IGPG HWPEVKQP YKVJ TGURGEV VQ RJ[UKQNQIKECN CPF FGXGNQROGPVCN EQPFKVKQPUÿ GPCDNGU VJG ENWUVGTKPI QH IGPGU QP VJG DCUKU QH EQTGURQPUKXGPGUUÿ CPF VJG CPCN[UKU QH INQDCN TGURQPUGU VQ OWVCVKQPU (5JCNQP GV CN. 199õþ øG 4KUK GV CN. 199ûþ óQFKEMC GV CN. 199ûþ UGG %JCRVGT 11). /KETQCTTC[ CPCN[UGU JCXG CNTGCF[ DGGP UWEEGUUHWNN[ CRRNKGF VQ VJG UVWF[ QH EJCPIGU KP INQDCN IGPG GZRTGUUKQP KP TGNCVKQP VQ ITQYVJ KP TKEJ XGTUWU OKPKOCN OGFKWOÿ FKCWZKE UJKHVÿ OGKQUKUÿ URQTWNCVKQPÿ RNQKF[ÿ EGNN E[ENGÿ IGPG OWVCVKQPÿ FTWI TGUKUVCPEGÿ UCNV UVTGUUÿ QUOQVKE UVTGUUÿ JGCV UVTGUUÿ QZKFCVKXG UVTGUUÿ GVJCPQN UVTGUUÿ QXGTGZRTGUUKQP QT CEVKXCVKQP QH VTCPUETKRVKQP HCEVQTUÿ CPF GZVGPFGF UGNGEVKQP HQT OWVCVKQPU (UGG EKVCVKQPU KP ôQHHGCW ù000þ 4QUU-/CEFQPCNF ù000). îG[ SWGUVKQPU VJCV TGNCVG VQ IGPG HWPEVKQP CTG: YJCV RTQVGKP FQGU CP WPMPQYP ú4ò GPEQFGÿ JQY GHHKEKGPVN[ KU VJG VTCPUETKDGF OGUUCIG VTCPUNCVGFÿ FQGU VJG GPEQFGF RTQVGKP KPVGTCEV YKVJ QVJGT RTQVGKPU QH MPQYP HWPEVKQP CPFöQT FQGU KV WPFGTIQ RQUV-VTCPUNCVKQPCN OQFKHKECVKQP KP TGURQPUG VQ EJCPIKPI RJ[UKQNQIKECN UVCVWU QT GPXKTQPOGPVCN EQPFKVKQPUÿ JQY SWKEMN[ KU VJG RTQVGKP FGITCFGFÿ CPF YJCV KU VJG UVGCF[ UVCVG NGXGN QH VJG RTQVGKP KP C EGNN WPFGT IKXGP EQPFKVKQPU! *GTGÿ VJG VGEJPKSWGU HQT CPCN[\KPI VJG RTQVGKP EQORNGOGPV QH EGNNU WPFGT IKXGP EKTEWOUVCPEGU (RTQVGQOKEU)ÿ CPF RTQVGKP-RTQVGKP KPVGTCEVKQP (VYQ-J[DTKF CPCN[UGU) CTG KORQTVCPV EQORNGOGPVU VQ KP UKNKEQÿ OWVCVKQPCN CPF IGPG GZRTGUUKQP UVWFKGU. óJKNUV VJGTG CTG UVKNN OCP[ VGEJPKECN NKOKVCVKQPU UWTTQWPFKPI RTQVGQOKEUÿ VJKU VGEJPQNQI[ KU FGXGNQRKPI TCRKFN[ CPF JCU DGEQOG CP KORQTVCPV CFjWPEV VQ IGPQOKE UVWFKGU. 4GNCVKXG VQ IGPG GZRTGUUKQP CPCN[UGUÿ RTQVGKP UVWFKGU ECP TGXGCN RQUV-VTCPUNCVKQPCN CNVGTCVKQPU UWEJ CU RJQURJQT[NCVKQP UVCVGU QH UQOG RTQVGKPUÿ YJKEJ CTG KORQTVCPV HQT CEVKXCVKQPö FGCEVKXCVKQP QH CEVKXKVKGU. /QTGQXGT VTCPUETKRV NGXGNU FQ PQV CNYC[U TGHNGEV VJG HKPCN NGXGN QH CEVKXKV[ RTQXKFGF D[ CP GPEQFGF RTQVGKPÿ UWEJ VJCV RTQVGQOKEU OC[ DG OQTG TGNGXCPV VQ VJG EGNN'U DKQNQIKECN UVCVWU VJCP VTCPUETKRVQOKEU. 2TQVGQOKEU JCU DGGP CRRNKGF VQ UVWF[ EJCPIGU KP RTQVGKP EQORNGOGPVU VJTQWIJ XCTKQWU ITQYVJ EQPFKVKQPUÿ FKCWZKE UJKHVÿ GZRQUWTG QH EGNNU VQ JGCV UJQEM (ôCTTGNU GV CN. 199û)ÿ J[FTQIGP RGTQZKFG (ôQFQP GV CN. 199÷)ÿ CPF QUOQVKE UVTGUU ($NQODGTI 199û). ;GV CPQVJGT KORQTVCPV CTGC QH IGPG HWPEVKQPCN FKUEQXGT[ KU VJG FGVGTOKPCVKQP QH YJGVJGT RTQVGKPU KPVGTCEV YKVJ QPG-CPQVJGT. +H VJG RTQFWEV QH CP WPMPQYP ú4ò KU KFGPVKHKGF CU KPVGTCEVKPI YKVJ C MPQYP RTQVGKP VJKU IKXGU C OCjQT ENWG CU VQ KVU DKQNQIKECN HWPEVKQP. 6JG HCXQWTGF OGVJQF HQT CPCN[\KPI RQVGPVKCN RTQVGKP-RTQVGKP KPVGTCEVKQPU KP [GCUV KU VJG VYQ-J[DTKF CUUC[ (òKGNFU CPF 5QPI 19÷9). 7GV\ GV CN. (ù000) RWDNKUJGF VJG HKTUV CTTC[-DCUGF VYQ-J[DTKF UETGGP QH C YJQNG RTQVGQOG FGUETKDKPI VJG VGUVKPI QH QXGT 1 Z 10õ RQUUKDNG KPVGTCEVKQPU TGUWNVKPI KP ù÷1 RQVGPVKCN RTQVGKP-RTQVGKP EQODKPCVKQPU. +VQ GV CN. (ù000ÿ ù001) CVVGORVGF VQ VGUV CNN RTQVGKPRTQVGKP RCKTU CPF KFGPVKHKGF CDQWV íñ00 RWVCVKXG KPVGTCEVKQPU YKVJ ÷í1 DGKPI JKIJN[ TGNKCDNG. 6YQ-J[DTKF CPCN[UGU JCXG CNUQ DGGP WUGF VQ FGVGTOKPG RTQVGKP KPVGTCEVKQPU KP O4ý# OGVCDQNKUO RCVJYC[U (òTQOQPV-4CEKPG GV CN. ù000). +P /C[ ù00ùÿ VJG /+25 %;ôø KPFKECVGF VJGTG YGTG CV NGCUV 9ûñ0 RQVGPVKCN RTQVGKP-RTQVGKP KPVGTCEVKQPU KP [GCUV. ôGPG ENWUVGT CPCN[UKU KU C EWTTGPVN[ HCXQWTGF CRRTQCEJ WUGF VQ KORN[ HWPEVKQPCNKV[ ($CPGTjGG CPF <JCPI ù00ùþ 6COCOGU GV CN. ù00ù). #TTC[ CPCN[UGU ECP FGHKPG
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
UGVU QT ENWUVGTU QH IGPGU YKVJ UKOKNCT GZRTGUUKQP RCVVGTPU. 6JG CUUWORVKQP KU VJGP OCFG VJCV VJGUG ENWUVGTU CTG TGNCVGF D[ VJGKT KPXQNXGOGPV KP RCTVKEWNCT DKQNQIKECN RTQEGUUGU (LQEMJCTV GV CN. 199õ). 6JG RTQEGUU QH FGHKPKPI VJG EGNNWNCT TQNG QH VJG ENWUVGTU V[RKECNN[ KPXQNXGU KP UKNKEQ CPCN[UKU D[ TGHGTGPEG VQ FKHHGTGPV FCVCDCUGU. óW GV CN. (ù00ù) JCXG FGOQPUVTCVGF VJG WUG QH ENWUVGT CPCN[UGU VQ KFGPVKH[ PGY IGPG OGODGTU QH GZKUVKPI HWPEVKQPCN ECVGIQTKGU KPENWFKPI ù÷ñ ECPFKFCVG RTQVGKPU KPXQNXGF KP VTCPUETKRVKQPÿ RTQEGUUKPI CPF VTCPURQTV QH PQP-EQFKPI 4ý# OQNGEWNGU. 5GXGTCN QH VJGUG ECPFKFCVGU CRRGCT VQ DG KPXQNXGF KP TKDQUQOCN 4ý# RTQEGUUKPI. %QORWVCVKQPCN CPCN[UKU QH QXGT ùû00 RWVCVKXG RTQVGKP-RTQVGKP KPVGTCEVKQPU FGTKXGF HTQO VYQ-J[DTKF FCVCDCUGU TGXGCNGF C NCTIG PGVYQTM QH ù3ñ÷ KPVGTCEVKQPU COQPI 1ñí÷ RTQVGKPU (5EJYKMQYUMK GV CN. ù000). 2TQVGKPU QH MPQYP HWPEVKQP CPF EGNNWNCT NQECVKQPU YGTG ENWUVGTGF VQIGVJGT YKVJ õ3ð QH KPVGTCEVKQPU QEEWTTKPI DGVYGGP RTQVGKPU YKVJ C EQOOQP HWPEVKQPCN CUUKIPOGPV CPF ûõð DGVYGGP RTQVGKPU KP VJG UCOG UWD-EGNNWNCT EQORCTVOGPV. #U QH ,CPWCT[ ù00ù VJG /+25 %;ôø TGRQTVGF VJCV í0ð QH VJG S. cerevisiae IGPGU CTG OGODGTU QH HCOKNKGUÿ CPF 39õ3 ú4òU OCVEJGF VQ CV NGCUV QPG HWPEVKQPCN ECVGIQT[. /CP[ QH VJG ú4òU YGTG CUUKIPGF VQ OQTG VJCP QPG HWPEVKQPCN ECVGIQT[ÿ KPFKECVKPI EQPUKFGTCDNG QXGTNCR QH EGNNWNCT HWPEVKQPUÿ YJKEJ KU PQV UWTRTKUKPI. # VQVCN QH 10õõ IGPGU JCXG DGGP CUUKIPGF VQ DTQCF OGVCDQNKUO (G.I. COKPQ CEKFÿ PKVTQIGPÿ UWNRJWTÿ PWENGQVKFGÿ RJQURJCVGÿ ECTDQP EQORQWPFÿ NKRKFÿ HCVV[ CEKF CPF KUQRTGPQKFÿ XKVCOKPUÿ EQHCEVQTU CPF RTQUVJGVKE ITQWRUÿ CPF UGEQPFCT[ OGVCDQNKUO HWPEVKQP). ©PGTI[ OGVCDQNKUO-CUUQEKCVGF ú4òU VQVCNGF ùñùÿ EGNN E[ENG CPF øý# RTQEGUUKPI õù÷ÿ VTCPUETKRVKQP ûû1ÿ RTQVGKP U[PVJGUKU 3ñ9ÿ RTQVGKP HQNFKPIÿ OQFKHKECVKQP CPF FGUVKPCVKQP ñ9ñÿ EGNNWNCT VTCPURQTV CPF VTCPURQTV OGEJCPKUOU í9ñÿ EGNNWNCT EQOOWPKECVKQPö UKIPCN VTCPUFWEVKQP OGEJCPKUOU ñ9ÿ EGNN TGUEWGÿ FGHGPUG ùû÷ÿ TGIWNCVKQP QHö KPVGTCEVKQP YKVJ EGNNWNCT GPXKTQPOGPV 199ÿ EGNN HCVG íùûÿ VTCPURQUCDNG GNGOGPVUÿ XKTCN CPF RNCUOKF RTQVGKPU 11õÿ EQPVTQN QH EGNNWNCT QTICPK\CVKQP ù09ÿ UWDEGNNWNCT NQECNK\CVKQP ùùñ÷ÿ RTQVGKP CEVKXKV[ TGIWNCVKQP 13ÿ RTQVGKP QT NKRKF DKPFKPI íÿ CPF VTCPURQTV HCEKNKVCVKQP 313. #DQWV í0ð (ùñ1í) QH ú4òU YGTG UVKNN HWPEVKQPCNN[ WPENCUUKHKGFÿ QT ENCUUKHKECVKQP YCU WPENGCT. 2.2.2 )GPQOGU QH KPFWUVTKCN Saccharomyces CPF PQP-Saccharomyces [GCUVU $TGYKPI CPF FKUVKNNKPI [GCUVU CTG YKVJ TCTG GZEGRVKQPUÿ URGEKGU QH Saccharomyces. /QNGEWNCT CPCN[UGU QH 9÷ KUQNCVGU ITQWRGF VJGUG [GCUVU YKVJKP S. cerevisiaeÿ S. uvarumÿ S. carlsbergensisÿ S. bayanus CPF S. monacensis (2GFGTUGP 19÷ñÿ 19÷õ). S. carlsbergensis KU RTQDCDN[ C J[DTKF QH S. cerevisiae CPF S. monacensis (îKGNNCPF-$TCPFV GV CN. 199ñþ *CPUGP CPF îKGNNCPF-$TCPFV 199õþ UGG %JCRVGT ñ). 6JGTG CRRGCTU VQ JCXG DGGP C JKIJ NGXGN QH KPVGTURGEKHKE ETQUUKPI DGVYGGP Saccharomyces UGPUW UVTKEVQ URGEKGU (FG $CTTQU LQRGU GV CN. ù00ù). 5VTCKPU QH YKPG [GCUVÿ EKFGT [GCUV CPF S. bayanus UJQY PWENGCT IGPQOG UKOKNCTKVKGU VQ DQVJ S. cerevisiae CPF QVJGT S. bayanus-NKMG UVTCKPUÿ CPF VYQ NCIGT UVTCKPU YGTG HQWPF VQ DG J[DTKFU QH S. cerevisiae CPF S. pastorianus (FG $CTTQU LQRGU GV CN. ù00ù).
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ù00ù). øKRNQKF UVTCKPU QH C. tropicalis RQUUGUU 10-1ù EJTQOQUQOGU CPF JCXG IGPQOGU QH DGVYGGP ù0 CPF 31 /D ($NCPFKP GV CN. ù000). Y. lipolytica KUQNCVGU UJQY EJTQOQUQOCN NGPIVJ RQN[OQTRJKUOU DWV KP IGPGTCN VGTOU VJKU [GCUV JCTDQWTU HQWT VQ UKZ EJTQOQUQOGU TCPIKPI HTQO VYQ VQ CNOQUV UKZ /D KP UK\G HQT C VQVCN IGPQOG QH CDQWV 1ù VQ ùù /D ($CTVJ CPF ôCKNNCTFKP 199ûþ %CUCToIQNC GV CN. 199ûÿ ù000þ UGG %JCRVGT ÷). îCT[QV[RKPI QH 19 H. anomala KUQNCVGU TGXGCNGF HQWT ITQWRU (ýCWOQX GV CN. ù001). %JTQOQUQOG UK\GU TCPIGF HTQO ÷ñ0 MD VQ 3.ñ /Dÿ YKVJ CNN UVTCKPU ECTT[KPI C EJTQOQUQOG QH CDQWV 1.ù /D. *CRNQKF EJTQOQUQOG PWODGT YCU PKPG VQ 11 CPF EJTQOQUQOG NGPIVJ RQN[OQTRJKUOU YGTG EQOOQP. Hanseniaspora URGEKGU GZJKDKV DGVYGGP UKZ CPF GKIJV EJTQOQUQOGUÿ TCPIKPI HTQO ûñ0 MD VQ ù.õ /Dÿ CPF Kloeckera RQUUGUUGU UKZ EJTQOQUQOGU TCPIKPI HTQO û90 MD VQ ù.í /D (%CFG\ GV CN. ù00ù). ôGPG QTFGT CPCN[UKU QH FCVC UVGOOKPI HTQO VJG ôoPQNGXWTGU UWTXG[ QH JGOKCUEQO[EGVQWU [GCUVU (UGG 5QWEKGV GV CN. ù000ÿ CPF VJG ù0 CUUQEKCVGF RCRGTU FGUETKDKPI MG[ FCVC UVGOOKPI HTQO VJG UWTXG[) KPFKECVGU VJCV IGPG CTTCPIGOGPVU KP K. lactisÿ K. marxianusÿ K. thermotoleransÿ S. kluyveri CPF Z. rouxii CTG EQPUKUVGPV YKVJ FKXGTIGPEG QH VJGUG [GCUVU HTQO S . cerevisiae DGHQTG RQN[RNQKFK\CVKQP QH VJG cerevisiae IGPQOG QEEWTTGF (óQPI GV CN. ù00ù). $[ EQPVTCUV C. glabrataÿ S. exiguus CPF S. servazzii UJQY JKIJGT EQPUGTXCVKQP QH IGPG CFjCEGPE[ KPFKECVKXG QH URNKVVKPI HTQO VJG S. cerevisiae NKPGCIG RQUV-RQN[RNQKFK\CVKQP. # TGRTGUGPVCVKXG QH S. bayanus XCT. uvarumÿ YJKEJ QEEWTU KP NQYGT VGORGTCVWTG YKPG HGTOGPVCVKQPUÿ UJQYGF ENQUG TGNCVGFPGUU VQ S. cerevisiae YKVJ PGCTN[ ù÷00 JQOQNQIQWU ú4òUÿ DWV CNUQ YKVJ ú4òU FWRNKECVGF KP S. uvarum DWV PQV S. cerevisiae ($QP GV CN. ù000). +P FKTGEV EQORCTKUQP YKVJ S. cerevisiaeÿ 6[1-NKMG TGVTQVTCPURQUQPU JCXG DGGP HQWPF KP S. uvarumÿ S. exiguusÿ S. servazzii CPF S. kluyveriÿ K. lactis CPF K. marxianus DWV PQV K. thermotolerans QT Z. rouxii (ýGWXGINKUG GV CN. ù00ù). S. cerevisiae 6[íNKMG GNGOGPVU QPN[ CRRGCTGF KP S. uvarumÿ YJGTGCU 6[ñ-NKMG GNGOGPVU YGTG HQWPF KP S. exiguusÿ P. angusta (H. polymorpha) CPF Debaryomyces hansenii. 2.2.3 (WPEVKQPCN CPCN[UGU QH KPFWUVTKCNN[ TGNGXCPV [GCUVU /QUV QH VJG CXCKNCDNG INQDCN FCVC QP HWPEVKQPCN IGPGVKEU TGHGTU VQ NCDQTCVQT[ UVTCKPU QH [GCUV ITQYP KP DCVEJ EWNVWTG (5GEVKQP ù.1). *QYGXGTÿ UQOG FCVC KU PQY GOGTIKPI DCUGF QP KPFWUVTKCN [GCUV UVTCKPU. òQT GZCORNGÿ OKETQCTTC[ CPCN[UKU QH [GCUV UVTCKPU VJCV UJQYGF HTGG\G-VJCY TGUKUVCPEG TGNGXCPV VQ HTQ\GP FQWIJ CRRNKECVKQPUÿ KPFKECVGF FKHHGTGPEGU KP GZRTGUUKQP QH UGXGTCN ú4òU KPXQNXGF KP EGNNWNCT QTICPK\CVKQPÿ OGVCDQNKUOÿ KPVTCEGNNWNCT VTCPURQTVÿ VTCPURQTV HCEKNKVCVKQPÿ EGNN ITQYVJ CPF FKXKUKQPÿ TGUEWG CPF FGHGPUGÿ VTCPUETKRVKQPÿ GPGTI[ OGVCDQNKUO CPF KQPKE JQOQGQUVCUKU (6CPIJG GV CN. ù000). 5VWFKGU QH IGPG GZRTGUUKQP KP EQPFKVKQPU TGNGXCPV VQ YKPG HGTOGPVCVKQPU (UGG 2oTG\-úTVsP GV CN. ù00ù) TGXGCNGF VJCV IGPGU KPXQNXGF KP COKPQ CEKF DKQU[PVJGUKU CPF RWTKPG DKQU[PVJGUKU IGPGTCNN[ ICXG JKIJ GZRTGUUKQP NGXGNU. 6JG VTCPUETKRVQOG QH C S. carlsbergensis NCIGT UVTCKP CPCN[\GF VJTQWIJQWV C RTQFWEVKQP-UECNG DGGT HGTOGPVCVKQPÿ UJQYGF VJCV CXGTCIG IGPG GZRTGUUKQP TCRKFN[ KPETGCUGF VQ C OCZKOWO VYQ FC[U CHVGT KPQEWNCVKQP (úNGUGP GV CN. ù00ù). ôGPG VTCPUETKRVKQP FGETGCUGF CU UWICT YCU EQPUWOGF. ôGPGU KPXQNXGF KP RTQVGKP CPF NKRKF
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VJG 10 EJCTCEVGTKUVKEU VGUVGF CPF ICXG UKOKNCT NGXGNU QH URQTWNCVKQP CPF URQTG XKCDKNKV[ RGTJCRU KPFKECVKPI EQOOQP CPEGUVT[ (,QJPUVQP GV CN. ù000). 5VWFKGU QH S. cerevisiae KUQNCVGU HTQO PCVWTCN YKPG HGTOGPVCVKQPU (K.G. TGN[KPI QP PCVWTCNN[ QEEWTTKPI [GCUVU TCVJGT VJCP QP UVCTVGT EWNVWTG UVTCKPU) JCXG TGXGCNGF VJG FGRVJ QH FKXGTUKV[ QH "PCVWTCN UVTCKP" IGPGVKEU. ôGPGVKE CPCN[UKU EQODKPGF YKVJ TGUVTKEVKQP HTCIOGPV NGPIVJ RQN[OQTRJKUOU FKUETKOKPCVGF 3í QWV QH 3õ PCVWTCN YKPG [GCUV KUQNCVGU (%CXCNKGTK GV CN. 199÷). úH ù39 UVTCKPU VGUVGF D[ VGVTCF CPCN[UKUÿ 1ññ JCF QPG QT OQTG JGVGTQ\[IQUKVKGU CPF ÷í CRRGCTGF VQ DG EQORNGVGN[ JQOQ\[IQWU HQT XCTKQWU VTCKVU (/QTVKOGT ù000). "ýCVWTCN" RCNO YKPG [GCUVU VJCV KPENWFG UVTCKPU QH Saccharomycesÿ Candidaÿ Schizosaccharomyces CPF Kluyveromyces UJQYGF XCTKCDNG IGPGVKE RTQRGTVKGU (©\GTQP[G CPF úMGTGPVWIDC ù001). /CP[ QH VJG UVTCKPU YGTG FKRNQKF CPF UGXGTCN URQTWNCVGF RTQFWEKPI HQWT URQTGF CUEK EQPVCKPKPI HQWT XKCDNG URQTGU. úH 109 UVTCKPU QH DTGYKPI Saccharomyces URGEKGUÿ ñ0 YGTG JGVGTQVJCNNKE CPF QH VJGUG UKZ YGTG FKRNQKFÿ VJTGG VTKRNQKFÿ 1ñ VGVTCRNQKFÿ GKIJV RGPVCRNQKFÿ UGXGP JGZCRNQKF CPF VJG TGOCKPFGT WPMPQYP (TGXKGYGF D[ ,QJPUVQP CPF úDGTOCP 19û9). +P HCEVÿ OQUV DGGT [GCUVU CTG QH WPMPQYP RNQKF[ CPF JCXG GZVTGOGN[ NQY URQTWNCVKQP HTGSWGPEKGU CPF NQY URQTG XKCDKNKVKGU (,QJPUVQP 1990). úHVGP URQTGU VJCV IGTOKPCVG CTG GKVJGT JQOQVJCNNKE QT UVGTKNG YKVJ TGURGEV VQ OCVKPI. #PCN[UKU QH UGXGP S. cerevisiae CNG [GCUVU TGXGCNGF VJCV HQWT URQTWNCVGF VQ RTQFWEG C CPF ½ V[RGUÿ QPG ICXG QPN[ ½ V[RG CPF VYQ RTQFWEGF PQP-OCVKPI URQTGU (,QJPUVQP CPF úDGTOCP 19û9). +V YCU WPENGCT YJGVJGT VJGUG URQTGU ECTTKGF UVGTKNKV[ OWVCVKQPU. #OQPI UGXGP S. carlsbergensis NCIGT [GCUVU HQWT URQTWNCVGF RQQTN[ QT PQV CV CNNÿ VYQ RTQFWEGF UVGTKNG URQTGU CPF QPG RTQFWEGF URQTGU VJCV OCVGF YKVJ DQVJ C CPF ½ V[RGU UVTCKPU (,QJPUVQP CPF úDGTOCP 19û9). 6JGUG CPCN[UGU UGTXG VQ KPFKECVG VJG EQORNGZKV[ QH DTGYGT'U [GCUV IGPGVKEU. $CMGT'U [GCUVUÿ YJKEJ CTG OQUVN[ UVTCKPU QH S. cerevisiae CNUQ GZJKDKV EQORNGZ IGPGVKE VTCKVU. 6JG[ GZJKDKV IGPGVKE HGCVWTGU EQPUKUVGPV YKVJ DGKPI FKRNQKFÿ VTKRNQKFÿ VGVTCRNQKFÿ RQN[RNQKF QT CPGWRNQKF (,QJPUVQP CPF úDGTOCP 19û9). +V KU QHVGP TGRQTVGF VJCV KPFWUVTKCN DCMGT'U [GCUV UVTCKPU CTG RQQT CV URQTWNCVKQP. *QYGXGT YG JCXG HQWPF VJCV URQTWNCVKQP HTGSWGPEKGU XCT[ EQPUKFGTCDN[ DGVYGGP DCMGT'U [GCUV UVTCKPUÿ TCPIKPI HTQO JKIJ HTGSWGPEKGU (>ñ0ð) VQ \GTQ URQTWNCVKQP TCVGU. øGRGPFKPI WRQP VJG UQWTEG UVTCKPÿ VJG PWODGT QH XKCDNG URQTGU XCTKGU YKFGN[. +P VGVTCF CPCN[UKU QH QPG EQOOQPN[ WUGF KPFWUVTKCN DCMGT•U [GCUV UVTCKPÿ YG QDUGTXGF VJCV PQ EQORNGVG VGVTCFU YGTG QDVCKPGF QWV QH VGP GZCOKPGF. úPEG IGTOKPCVGFÿ QPN[ C RTQRQTVKQP QH VJG URQTGU KUQNCVGF HTQO DCMGT•U [GCUV UVTCKPU FKURNC[ C OCVKPI V[RGÿ CPF VJKU OCVKPI V[RG KU PQV CNYC[U UVCDNGÿ PQT GZENWUKXG VQ QPG QT QVJGT V[RG. +P RCTVKEWNCTÿ CU GZRGEVGF HTQO C VGVTCRNQKF RCTGPVCN UVTCKPÿ C JKIJ RTQRQTVKQP QH IGTOKPCVGF URQTGU YKNN PQV JCXG C OCVKPI V[RG DGECWUG VJG[ JCXG C DCNCPEGF Cö½ IGPGVKE OCMG WR. 5QOGVKOGU VJG PQP-OCVKPI V[RG URQTGU ECP DG URQTWNCVGF HWTVJGT VQ IGPGTCVG VTWG JCRNQKFU. 6JG OCVKPI V[RG QH IGTOKPCVGF URQTGU QH KPFWUVTKCN DCMGT'U [GCUV UVTCKPU ECP UYKVEJ CV C YKFG TCPIG QH HTGSWGPEKGU. /CP[ IGTOKPCVGF URQTGU CTG CNOQUV RWTGN[ JGVGTQVJCNNKEÿ DWV C UKIPKHKECPV RTQRQTVKQP TCPIG HTQO UNKIJVN[ JQOQVJCNNKE VQ CNOQUV EQORNGVGN[ JQOQVJCNNKE. +H C IGTOKPCVGF [GCUV JCU C OCVKPI V[RGÿ VJG HTGSWGPE[ QH OCVKPI ECP TCPIG HTQO JKIJN[ GHHKEKGPV VQ GZVTGOGN[ RQQT.
30
2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
òKPCNN[ÿ UQOG JCRNQKFU JCXG C VGPFGPE[ VQ NQUG VJGKT OCVKPI V[RGU CHVGT GZVGPFGF UWDEWNVWTKPI. óJGTGCU K. marxianusÿ P. angusta/ H. polymorpha CPF Z. rouxii CTG JQOQVJCNNKEÿ PCVWTCN KUQNCVGU QH Y. lipolytica CTG EQOOQPN[ JGVGTQVJCNNKE. øKRNQKF Y. lipolytica UVTCKPU URQTWNCVG QP UQNKF QT KP NKSWKF OGFKWO CPF FQ PQV TGSWKTG PKVTQIGP-NKOKVCVKQP VQ FQ UQ ($CTVJ GV CN.199û). ©HHKEKGPEKGU QH URQTWNCVKQP CTG OGFKWOCPF UVTCKP-URGEKHKE CPF CUEQURQTG XKCDKNKV[ KU XCTKCDNG. úRRQUKVG OCVKPI V[RGU CTG FGUKIPCVGF GKVJGT MATA QT MATB ($CUUGN GV CN. 19û1). /CVKPI HTGSWGPEKGU QH PCVWTCN KUQNCVGU CPF HGTVKNKV[ QH J[DTKFU CTG NQY. *CRNQKF CPF FKRNQKF EGNNU QH Y. lipolytica CTG XGIGVCVKXGN[ UVCDNG. 6JG OKVQVKE ITQYVJ QH K. lactis KU RTGFQOKPCPVN[ JCRNQRJCUGÿ ECWUKPI FKRNQKF EGNNU VQ WPFGTIQ OGKQUKU URQPVCPGQWUN[ (5EJCHHTCVJ CPF $TGWPKI ù000). 5VWFKGU QH QTKIKPCN KUQNCVGU CPF OWVCIGPK\GF FGTKXCVKXGU QH H. polymorpha KPFKECVG VJKU [GCUV JCU RQVGPVKCN HQT IQQF OCVKPIÿ JKIJ HTGSWGPE[ QH URQTWNCVKQP CPF IQQF URQTG XKCDKNKV[ (LCJVEJGX GV CN. ù00ù). ôGPGVKE OCPKRWNCVKQPU GPCDNGF VJG KUQNCVKQP QH FGTKXCVKXGU QH FKRNQKF H. polymorpha VJCV YGTG WUGF VQ UJQY /GPFGNKCP UGITGICVKQP QH IGPGVKE OCTMGTU KP TCPFQO CPF VGVTCF CPCN[UGUÿ CNVJQWIJ KTTGIWNCT UGITGICVKQPU YGTG SWKVG EQOOQP (HKXG VQ ù0ð) RQUUKDN[ KPFKECVKPI CPGWRNQKFKGU. ýCWOQX GV CN. (ù001) EJCTCEVGTK\GF UGXGTCN UVTCKPU QH H. anomala. /QUV FKF PQV URQTWNCVG CV CNN QT URQTWNCVGF CV XGT[ NQY HTGSWGPEKGU. /CVKPI CEVKXKVKGU YGTG YGCM CPF CUEQURQTGU YGTG IGPGTCNN[ PQP-XKCDNG. /QUV J[DTKFU VJCV YGTG QDVCKPGF YGTG RQN[RNQKF. 5QOG OQPQURQTKE UVTCKPU HQTOGF FKRNQKF J[DTKFU YKVJ TGIWNCT UGITGICVKQPU QH CWZQVTQRJKE OCTMGTU.
2.3 +PFWUVTKCN [GCUV UVTCKP KORTQXGOGPV 2.3.1 +UUWGU CHHGEVKPI VJG EJQKEG QH IGPGVKE UVTCVGI[ 5VTCVGIKGU HQT IGPGVKE KORTQXGOGPV QH [GCUVU KPENWFG DTGGFKPI (CNUQ ECNNGF OCVKPI QT J[DTKFK\CVKQP)ÿ OWVCIGPGUKUÿ RTQVQRNCUVö URJGTQRNCUV HWUKQPÿ E[VQFWEVKQP QT Tøý# CRRTQCEJGU. óJGTG [GCUV ECP RTQFWEG TGCUQPCDNG PWODGTU QH UVCDNG OCVKPI V[RG URQTGUÿ DTGGFKPI KU CRRNKECDNGÿ YJGTGCU HQT [GCUV VJCV CTG CURQTQIGPQWUÿ QT RTQFWEG UVGTKNG URQTGUÿ OWVCIGPGUKU KU OQTG CRRNKECDNG. 2TQVQRNCUV HWUKQP KU CRRNKGF GURGEKCNN[ VQ KPVGTURGEKHKE CPF KPVGTIGPGTKE ETQUUGU. %[VQFWEVKQP KU WUGF VQ KPVTQFWEG E[VQRNCUOKE-KPJGTKVGF HCEVQTUÿ YJKNUV OCKPVCKPKPI VJG PWENGCT OCMG-WR QH C RCTGPVCN EGNN. %QODKPCVKQPU QH VJGUG CRRTQCEJGU CTG CNUQ WUGFÿ G.I. OWVCIGPGUKU CPF OCVKPI. %QODKPCVKQP QH Tøý# CPF ENCUUKECN OGVJQFU KU CNUQ C XKCDNG QRVKQP. 6JG CRRNKECVKQP QH Tøý# RTQXKFGU HQT JKIJN[ FKTGEVGF QWVEQOGU CPF RQVGPVKCNN[ TGFWEGF KPVGPUKV[ QH UETGGPKPI HQT KORTQXGF UVTCKPU. %QPUVTWEVGF UVTCKPU ECP DG IGPGVKECNN[ UVCDNG YKVJ RTGEKUG EJCPIGU DGKPI OCFG VQ VCTIGVGF IGPGU KPENWFKPI CNVGTCVKQP QH VJG EQPVTQN CPF UVTGPIVJ QH GZRTGUUKQP QH IGPGU QT VJG GNKOKPCVKQP QH WPFGUKTCDNG IGPG HWPEVKQPU. /CjQT NKOKVCVKQPU QH Tøý# CRRTQCEJGU CTG HKTUVN[ÿ VJG PGGF VQ MPQY YJCV IGPG(U) PGGF EJCPIKPI VQ CEJKGXG KORTQXGOGPVU KP FGUKTGF KPFWUVTKCNN[ TGNGXCPV RJGPQV[RGUÿ CPF UGEQPFN[ÿ VJG KUUWG QH CEEGRVCPEG QH "ôGPGVKECNN[ /QFKHKGF úTICPKUOU" (ô/úU). ýGKVJGT QH VJGUG OCVVGTU KU VTKXKCN. òWPEVKQPCN
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31
CPCN[UGU QH [GCUVU CTG RTQXKFKPI C OCUUKXG COQWPV QH FCVC KPFKECVKPI C JKIJ FGITGG QH EQORNGZKV[ KP IGPGVKE EQPVTQN QH [GCUV EGNN DKQNQI[. /CP[ IGPGU CTG ETQUUHWPEVKQPCN (QRGTCVKPI KP OQTG VJCP QPG KORQTVCPV EGNN HWPEVKQP)ÿ CPF OCP[ IGPGU CTG EQORGPUCVGF HQT D[ QVJGT IGPGU YJGP HWPEVKQPCNKV[ KU MPQEMGF QWV. 6JWUÿ KFGPVKH[KPI MG[ IGPGVKE EJCPIGU HQT Tøý# UVTCVGIKGU QH UVTCKP KORTQXGOGPV TGOCKPU C OCjQT EJCNNGPIG. +P HCEV OCP[ KPFWUVTKCNN[ KORQTVCPV VTCKVU CTG IQXGTPGF D[ EQODKPCVKQPU QH UGXGTCN IGPGU CPF VJKU OGCPU VJCV KP UQOG ECUGU Tøý# CRRTQCEJGU CTG UKORN[ PQV RTCEVKECNÿ NGCXKPI DTGGFKPI UVTCVGIKGU CU VJG DGUV QRVKQP. ©XGP YJGP UWKVCDNG Tøý# VCTIGVU CTG KFGPVKHKGF CPF GZRGTKOGPVCNN[ RTQXGPÿ VJG OCVVGT QH VJG ô/ú FGDCVG JCU UVKNN VQ DG FGCNV YKVJ. úHVGPÿ KTTCVKQPCN CPF GOQVKQPCN CTIWOGPVU CTG WUGF VQ OCKPVCKP C IGPGTCNN[ CPVK-ô/ú UVCPEG. 4GICTFNGUU QH CP[ UQEKQGEQPQOKE CFXCPVCIGU VQ DG ICKPGF HTQO KORTQXKPI UVTCKPU XKC Tøý# OGVJQFUÿ RWDNKE RGTEGRVKQP CPF NGIKUNCVKQP OCMG KV FKHHKEWNV VQ UGG ô/ú'U DGKPI WUGF EQOOGTEKCNN[ KP VJG PGCT HWVWTG. 6JKU KU GURGEKCNN[ UQ KP VJG HQQF CPF HGTOGPVCVKQP KPFWUVTKGUÿ YJGTG OCP[ KPFWUVTKCN [GCUVU GPjQ[ VJG KORQTVCPV UVCVWU QH DGKPI IGPGTCNN[ TGEQIPK\GF CU UCHG (ô4#5) CPF VJGTG KU VJG PGGF VQ OCKPVCKP VJKU UVCPFKPI CPF CXQKF VJG "òTCPMGPHQQF" NCDGN VJCV Tøý# UVTCKPU TKUM. %NCUUKECN OGVJQFU ECP NGCF VQ JKIJ IGPGVKE FKXGTUKV[ÿ IKXKPI VJG CDKNKV[ VQ EQODKPG QRVKOCN CNNGNGU CPF RQN[IGPKE VTCKVUÿ CPF GXGP VQ KPVGTIGPGTKEöKPVGTURGEKHKE GZEJCPIGU. /QUV UKIPKHKECPVN[ÿ ENCUUKECN IGPGVKE CRRTQCEJGU CTG CEEGRVCDNG KP VGTOU QH RWDNKE CPF RQNKVKECN TGNCVKQPUÿ GXGP VJQWIJ VJG VGEJPKSWGU NGCF VQ OWEJ DTQCFGT IGPGVKE EJCPIGU VJCP YQWNF DG UGGP XKC Tøý# VGEJPKSWGU. #PQVJGT KORQTVCPV CFXCPVCIG QH ENCUUKECN IGPGVKE CRRTQCEJGU KU VJCV VJGKT GORKTKECN PCVWTG QHVGP OGCPU VJGTG KU C NQY FGRGPFGPE[ QP RTKQT MPQYNGFIG QH IGPGVKE ECWUG CPF GHHGEV QH KPFWUVTKCNN[ TGNGXCPV VTCKVU. # OCjQT FKUCFXCPVCIG QH ENCUUKECN OGVJQFU KU VJCV QWVEQOGU CTG WUWCNN[ WPEQPVTQNNGF UWEJ VJCV VJG RTQDCDKNKV[ QH CEJKGXKPI VJG EQTTGEV EQODKPCVKQP QH IGPGUÿ CNNGNGU QT OWVCVKQPU KU JKIJN[ FGRGPFGPV QP VJG PWODGTU QH OWVCPVUÿ HWUKQP RTQFWEVU QT J[DTKFU VJCV CTG RTQFWEGF CPF UETGGPGF. 2.3.2 /CVKPI /CVKPI QT DTGGFKPI QH NCDQTCVQT[ UVTCKPU QH S. cerevisiae KU TGNCVKXGN[ UKORNG DGECWUG VJGUG UVTCKPU JCXG UVTCKIJVHQTYCTF IGPGVKE RTQRGTVKGU VJCV NCTIGN[ HQNNQY /GPFGN'U RTGFKEVKQPU HQT IGPGVKE UGITGICVKQP (5GEVKQP ù.3). #U CP GZCORNGÿ EQPUKFGT VJCV CP KORQTVCPV RJGPQV[RG KU EQPVTQNNGF D[ VYQ IGPGU (# CPF $) CPF VJCV UVTQPI (#ÿ $) CPF YGCM (Cÿ D) CNNGNGU QH VJGUG VYQ IGPGU GZKUV YKVJKP FKRNQKF UVTCKPU (DQVJ #Cö$D). +H VYQ FKRNQKF UVTCKPU CTG URQTWNCVGF VQ IKXG HQWT-URQTGF CUEKÿ CPF CNN URQTGU IGTOKPCVG VQ IKXG UVCDNG JGVGTQVJCNNKE OCVKPI V[RGUÿ VJGP VJGTG CTG CV NGCUV 1õ RQUUKDNG EQODKPCVKQPU HQT TGEQODKPKPI CNNGNGU QH VJG VYQ FKHHGTGPV IGPGU YJGP TGHQTOKPI FKRNQKFU. úPN[ QPG EQODKPCVKQP RTQXKFGU VJG FGUKTGF ##ö$$ IGPQV[RG. +P VJKU ECUG VJGTG KU C QPG KP 1õ EJCPEG QH UWEEGUU KP C DTGGFKPI UVTCVGI[. *QYGXGTÿ VJG PWODGT QH IGPGU KPXQNXGF KP EQPHGTTKPI VJG FGUKTGF EJCTCEVGTKUVKEU TGSWKTGF KP CP KPFWUVTKCN UVTCKP KU WUWCNN[ WPMPQYP. òWTVJGTOQTGÿ VJG PWODGT QH EQODKPCVKQPU
2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
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(KI. þ. 7UG QH HNWQTGUEGPEG CPF (#%5 VQ QDVCKP GPTKEJGF URQTG RQRWNCVKQPU. # URQTWNCVKPI EWNVWTG QH KPFWUVTKCN DCMGT'U [GCUV YCU UVCKPGF YKVJ C HNWQTGUEGKP-DCUGF F[G. 8GIGVCVKXG EGNNU GZRGN VJG F[G YJGTGCU URQTGU TGVCKP VJG F[G. 6JG RQRWNCVKQP YCU UWDjGEVGF VQ HNQY E[VQOGVT[ CPF HNWQTGUEGPEG CEVKXCVGF EGNN UQTVKPI (ò#%5). LGHV RCPGN UJQYU VJG FKUVTKDWVKQP QH XGIGVCVKXG EGNNU CPF CUEK KP VJG KPKVKCN RQRWNCVKQP: 9ûð QH VJG RQRWNCVKQP JCF C HNWQTGUEGPV KPVGPUKV[ QH GSWCN VQ QT ITGCVGT VJCP 300 WPKVU. 4KIJV RCPGN UJQYU VJG FKUVTKDWVKQP QH XGIGVCVKXG EGNNU CPF CUEK KP VJG RQRWNCVKQP CHVGT ò#%5 YKVJ UGNGEVKQP HQT JKIJ HNWQTGUEGPV KPVGPUKV[: 90ð QH VJG RQRWNCVKQP JCF C HNWQTGUEGPV KPVGPUKV[ QH GSWCN VQ QT ITGCVGT VJCP 1ÿ000 WPKVU. /KETQUEQRKE CPCN[UKU QH VJG UQTVGF RQRWNCVKQP TGXGCNGF C JKIJ RWTKV[ QH CUEK.
QH CUUQTVGF IGPGU RQUUKDNG KP OGKQUKU KPETGCUGU GZRQPGPVKCNN[ YKVJ VJG PWODGT QH IGPGU IQXGTPKPI C VTCKV. 6JWU 10 IGPGU ECP DG TCPFQON[ CUUQTVGF CRRTQZKOCVGN[ QPG VJQWUCPF YC[U (ù10)ÿ ù0 IGPGU ECP DG TCPFQON[ CUUQTVGF KP C OKNNKQP YC[Uÿ CPF 30 IGPGU ECP DG CUUQTVGF KP C DKNNKQP YC[U. +H OQTG VJCP 10 IGPGU CHHGEV C RCTVKEWNCT EJCTCEVGTKUVKEÿ VJGP VJG PWODGT QH IGTOKPCVGF URQTGU VJCV PGGF VQ DG GZCOKPGF KPETGCUGU CV C FTCOCVKE TCVG. +P NKIJV QH VJGUG HCEVU KV KU UCNKGPV VQ TGOGODGT VJCV CP KPFWUVTKCN [GCUV UVTCKP TGSWKTGU VJG EQODKPCVKQP QH OCP[ EJCTCEVGTKUVKEU. 4GCNKUVKECNN[ÿ FGUKTGF IGPG EQODKPCVKQPU QEEWT CV HTGSWGPEKGU QH OWEJ NGUU VJCP QPG RGT OKNNKQPÿ OGCPKPI VJCV JKIJN[ GHHKEKGPV UETGGPKPI RTQEGFWTGU CTG PGGFGF. ýGXGTVJGNGUUÿ OCVKPI KU C WUGHWN OGCPU QH FGXGNQRKPI PQXGN KPFWUVTKCN [GCUVU YJGTG KPFWUVTKCN UVTCKPU URQTWNCVG CPF URQTGU IGTOKPCVG VQ IKXG XKCDNGÿ JGVGTQVJCNNKE CPF UGZWCNN[ EQORGVGPV OCVKPI V[RG UVTCKPU. # EQOOQP FKHHKEWNV[ KP QDVCKPKPI NCTIG PWODGTU QH KPFWUVTKCN [GCUV URQTGU YKVJ YJKEJ VQ ECTT[QWV OCVKPI RTQITCOOGU CTKUGU VJTQWIJ KPCDKNKV[ VQ GNKOKPCVG RCTGPVCNÿ XGIGVCVKXG (WPURQTWNCVGF) EGNNU YKVJQWV FCOCIKPI VJG URQTGU. 6JKU KU C RCTVKEWNCT RTQDNGO YJGTG XGT[ NQY URQTWNCVKQP HTGSWGPEKGU QEEWT. 6JG RTGUGPEG QH VJG RCTGPVCN EGNN V[RGU ECP EQPHQWPF OCVKPI GZRGTKOGPVU CPF VJGKT TGOQXCN D[ UQNXGPVÿ JGCV QT J[FTQRJQDKE RJCUG UGRCTCVKQP ECP NGCF VQ KPXKCDKNKV[ QH UQOG URQTGU. óG JCXG QXGTEQOG VJKU RTQDNGO D[ WUKPI HNWQTGUEGPV UVCKPKPI CPF HNQY E[VQOGVT[
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
33
(HNWQTGUEGPEG CEVKXCVGF EGNN UQTVKPIþ ò#%5). +P VJKU CRRTQCEJÿ URQTGU CPF EGNNU CTG UVCKPGF YKVJ C HNWQTGUEGPV F[G VJCV KU TGVCKPGF D[ URQTGU DWV PQV XGIGVCVKXG EGNNU. 6JKU GPCDNGU VJG HNQY E[VQOGVGT VQ FGVGEV CPF FKUVKPIWKUJ URQTGU HTQO XGIGVCVKXG EGNNU. 6JG URQTGU ECP VJGP DG KUQNCVGF WUKPI ò#%5ÿ CV TCVGU GZEGGFKPI 1000 RGT UGEQPF (òKI. ù). *CXKPI QDVCKPGF JKIJ PWODGTU QH URQTGUÿ OCVKPI RTQITCOOGU ECP DG TCPFQO QT FKTGEVGF. 4CPFQO OCVKPI ECP KPXQNXG OCUU QT TCTG GXGPVU. +P OCUU OCVKPIÿ OCP[ VJQWUCPFU QH OCVKPI V[RG UVTCKPU CTG OKZGF YKVJQWV RTKQT UETGGPKPI. 6JKU CRRTQCEJ KU OQUV CRRNKECDNG KH OCVKPI HTGSWGPEKGU CTG GZRGEVGF VQ DG NQYÿ QT KH UWKVCDNG UETGGPU CTG WPCXCKNCDNG HQT URGEKHKE RJGPQV[RKE EJCTCEVGTK\CVKQP QH OCVKPI V[RG UVTCKPU. +P TCTG OCVKPIÿ KPFWUVTKCN UVTCKPU VJCV HCKN VQ UJQY OCVKPI V[RGU CV WUGHWN HTGSWGPEKGU CTG HQTEG-OCVGF YKVJ MPQYP OCVKPI V[RG UVTCKPU. 2TQFWEVU QH TCTG OCVKPI CTG QHVGP CDNG VQ URQTWNCVGÿ VJGTGD[ RTQXKFKPI C DCUKU HQT HWVWTG OGKQVKE J[DTKFK\CVKQPU (,QJPUVQP 1990). óJGVJGT FGTKXGF VJTQWIJ OCUU QT TCTG OCVKPIUÿ TCPFQO J[DTKFU CTG UGNGEVGF HTQO WPOCVGF EGNNU KP XCTKQWU YC[U (UGG DGNQY). øKTGEVGF OCVKPI KPXQNXGU VJG UETGGPKPI CPF EJCTCEVGTK\CVKQP QH UVCDNG OCVKPI V[RG UVTCKPU RTKQT VQ OCVKPIÿ YKVJ C XKGY VQ QDVCKPKPI URGEKHKE EQODKPCVKQPU QH VTCKVU. øKTGEVGF OCVKPI KU OQUVN[ CRRNKECDNG YJGTG IQQF UETGGPU CTG CXCKNCDNG VQ CUUGUU IGTOKPCVGF URQTGU. úWT GZRGTKGPEG YKVJ KPFWUVTKCN DCMGT'U [GCUVU KPFKECVGU VJCV RJGPQV[RGU XKVCN VQ KPFWUVTKCN WVKNKV[ ECP XCT[ EQPUKFGTCDN[ DGVYGGP URQTGU VJCV JCXG DGGP IGTOKPCVGF. 6JG[ XCT[ KP VJGKT ITQYVJ TCVGU QP INWEQUG DCUGF TKEJ OGFKWOÿ VJG[ XCT[ KP VJGKT ITQYVJ TCVGU CPF EGNN [KGNF QP CGTQDKE OGFKCÿ VJGKT TCVG QH HGTOGPVCVKQP QH OCNVQUGÿ VJGKT TCVG QH HGTOGPVCVKQP KP JKIJN[ QUOQVKE OGFKWO CPF KP OCP[ QVJGT EJCTCEVGTKUVKEU VJCV YG JCXG GZCOKPGF. #U CP GZCORNGÿ 6CDNG ù UJQYU UGITGICVKQP QH CGTQDKE ITQYVJ ECRCEKV[ CPF OCNVQUG-J[FTQN[\KPI (½INWEQUKFCUG) CEVKXKV[ KP FKUUGEVGF VGVTCFU QH C EQOOGTEKCN DCMGT'U [GCUV. %CRCEKV[ HQT GHHKEKGPV QT KPGHHKEKGPV CGTQDKE ITQYVJ UGITGICVGF ù:ù KPFKECVKPI VJCV VJKU VTCKV KU IQXGTPGF KP VJKU KPFWUVTKCN DCMGT'U [GCUV UVTCKP D[ JGVGTQ\[IQWU PWENGCT IGPGU TCVJGT VJCP OKVQEJQPFTKCN HWPEVKQPU. 6JG ½-INWEQUKFCUG CEVKXKVKGU FKF PQV UJQY UWEJ ENGCT UGITGICVKQP RCVVGTPU. +P VGVTCFU *1 CPF *õ VJGTG CRRGCTGF VQ DG C ù:ù UGITGICVKQP QH TGNCVKXGN[ JKIJ CPF NQY GP\[OG CEVKXKVKGUÿ CNVJQWIJ VJG CDUQNWVG XCNWGU XCTKGF DGVYGGP VGVTCFU. 6GVTCFU *ùÿ *3ÿ *í CPF *ñ ICXG URQTGU YKVJ CUUQTVGF NGXGNU QH ½-INWEQUKFCUG. 6JGUG FCVC KPFKECVG EQORNGZKV[ QH MAL IGPG QTICPK\CVKQP KP VJG RCTGPVCN UVTCKP RTQDCDN[ TGNCVGF VQ JGVGTQ\[IQUKVKGU KP OWNVKRNG MAL NQEK (UGG 5GEVKQP í). 6JG XCTKCDKNKV[ KP DTQCF EJCTCEVGTKUVKEU QH IGTOKPCVGF URQTGU TGHNGEVU VJG IGPGVKE NQCF KP VJG RCTGPVCN UVTCKP. òQT GZCORNGÿ KV KU QHVGP PQV ENGCT JQY CP KPFWUVTKCN UVTCKP YCU RTQFWEGFÿ CPF JQY NQPI KV JCU DGGP RTQRCICVGF KP CP CUGZWCN HCUJKQP. +P VJG ECUG QH UQOG [GCUV UVTCKPUÿ KV UGGOU NKMGN[ VJCV VJG[ JCXG DGGP IGPGTCVGF CU C TGUWNV QH C OWVCIGPGUKU RTQEGFWTGÿ CPF YKNN VJWU JCXG OCP[ IGPGVKE NGUKQPU VJCV CTG OCUMGF KP VJG FKRNQKFöVGVTCRNQKFöCPGWRNQKF DCEMITQWPFU. 7RQP URQTWNCVKQPÿ VJGUG NGUKQPU ECP OCMG VJGKT RTGUGPEG MPQYP D[ CHHGEVKPI OCP[ OGCUWTCDNG EJCTCEVGTKUVKEU. #U C TGUWNV QH VJG JKIJ FGITGG QH RJGPQV[RKE FKXGTUKV[ QDUGTXGF YKVJ URQTWNCVGF KPFWUVTKCN [GCUV UVTCKPUÿ C NCTIG PWODGT QH URQTG RTQIGP[ OWUV DG GZCOKPGF VQ KFGPVKH[ VJQUG VJCV CTG UWKVCDNG HQT WUG KP C OCVKPI RTQITCO. 6JG PWODGT QH URQTGU VJCV PGGF VQ DG UETGGPGF KU XCTKCDNG CPF FGRGPFU WRQP VJG RCTVKEWNCT IGPGVKE
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
6CDNG þ. 5GITGICVKQP QH CGTQDKE ITQYVJ CPF OCNVQUG-WVKNK\KPI CEVKXKV[ KP VGVTCFU QH C EQOOGTEKCN DCMGT'U [GCUV 6GVTCF *1C *1D *1E *1F *ùC *ùD *ùE *ùF *3C *3D *3E *3F *íC *íD *íE *íF *ñC *ñD *ñE *ñF *õC *õD *õE *õF
#GTQDKE ITQYVJ
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ôQQF ôQQF 2QQT 2QQT ôQQF 2QQT ôQQF 2QQT ôQQF ôQQF 2QQT 2QQT 2QQT ôQQF ôQQF 2QQT 2QQT ôQQF ôQQF 2QQT ôQQF 2QQT ôQQF 2QQT
0.õ0 0.í0 1.ñ0 1.30 1.õ0 0.íñ 0.ù9 0.÷3 0.1ù 1.ñ0 1.00 0.0ù 0.09 0.õ÷ 0.3÷ 0.ññ 1.õ0 0.í9 0.÷9 0.íû 0.ññ 0.1í 0.ñ9 0.1÷
OCMG-WR QH VJG RCTGPV UVTCKPÿ CPF KP RCTVKEWNCT VJG COQWPV QH JGVGTQ\[IQUKV[ KP VJG IGPGU TGSWKTGF HQT VJG KPFWUVTKCN EJCTCEVGTKUVKEU. 6JWU YG HKPF VJCV UQOG UVTCKPU QH DCMGT'U [GCUVU RTQXKFG URQTGU YKVJ EQODKPGF WUGHWN EJCTCEVGTKUVKEU CV TGNCVKXGN[ JKIJ HTGSWGPEKGU QH CDQWV 1 KP ñ00ÿ YJGTGCU QVJGT UVTCKPU RTQXKFG WUGHWN URQTGU CV XGT[ NQY HTGSWGPEKGU QH NGUU VJCP QPG KP 10í. óJGVJGT TCPFQO QT FKTGEVGF OCVKPI KU WUGFÿ J[DTKF UVTCKPU PGGF VQ DG KUQNCVGF HTQO RCTGPVCN QT OCVKPI V[RG UVTCKPU KP OKZGF EWNVWTGU. 7PNKMG NCDQTCVQT[ [GCUV UVTCKPUÿ KPFWUVTKCN [GCUV UVTCKPU FQ PQV IGPGTCNN[ JCXG CWZQVTQRJKE OCTMGTUÿ CPF RCTGPVCN UVTCKPUÿ JCRNQKFU CPF J[DTKFU CTG CNN CDNG VQ ITQY QP VJG UCOG OGFKC OCMKPI KV KORQUUKDNG VQ WUG UVCPFCTF EQORNGOGPVCT[ IGPGVKE OCTMGTU VQ KFGPVKH[ PQXGN J[DTKFU. 6JKU RTQDNGO KU GZCEGTDCVGF YJGTG OCVKPI V[RG UVTCKPU FQ PQV OCVG CV JKIJ HTGSWGPE[ CPF URQPVCPGQWUN[ NQUG VJGKT OCVKPI CDKNKV[. 5GXGTCN CNVGTPCVKXG OGVJQFU ECP DG WUGF VQ URGEKHKECNN[ KFGPVKH[ J[DTKFU. /QUV UKORN[ÿ VYQ UVTCKPU QH QRRQUKVG OCVKPI V[RG ECP DG OKZGF VQIGVJGTÿ KPEWDCVGF HQT UGXGTCN FC[Uÿ CPF VJGP UVTGCMGF QWV VQ UKPING EQNQPKGU. 2TQXKFGF VJG OCVKPI TGCEVKQP YCU GHHKEKGPVÿ CPF VJG OCVKPI V[RGU QH VJG RCTGPVCN JCRNQKFU YGTG UVCDNGÿ J[DTKF UVTCKPU ECP DG TGCFKN[ KFGPVKHKGF HTQO VJG UKPING EQNQPKGU D[ UETGGPKPI HQT OCVKPI V[RG. +H QNKIQPWENGQVKFG UGSWGPEGU CTG CXCKNCDNG VJCV ECP WPKSWGN[ KFGPVKH[ DQVJ RCTGPVCN URQTG V[RGU HTQO
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QHHURTKPIÿ VJG PCVWTG QH VJG RWVCVKXG J[DTKFU ECP DG EQPHKTOGF D[ IGN GNGEVTQRJQTGUKU QH 2%4 RTQFWEVU. *QYGXGTÿ RTQDNGOU QEEWT KH VJG URQTG V[RGU CTG PQV ENGCTN[ FKUVKPIWKUJCDNG D[ 2%4ÿ QT KH OCVKPI V[RG YCU PQV UVCDNG QT OCVKPI QEEWTTGF CV XGT[ NQY HTGSWGPEKGU. # HWTVJGT OGVJQF VQ QXGTEQOG VJG NCEM QH CWZQVTQRJKE OCTMGTU KU VQ KPVTQFWEG FKHHGTGPV EJTQOQUQOCN QT OKVQEJQPFTKCN CPVKDKQVKE TGUKUVCPEGU KPVQ GCEJ QH VJG RCTGPVCN [GCUV UVTCKPUÿ CPF WUG FWCN CPVKDKQVKE TGUKUVCPEG VQ KFGPVKH[ J[DTKF UVTCKPU (2WVTCOGPV GV CN. 19û3). 6JG OCjQT NKOKVCVKQP VQ VJKU OGVJQF KU VJG UOCNN TCPIG QH UWKVCDNG OCTMGTU CPF VJG NCDQWT KPVGPUKXG PCVWTG QH VJG RTQEGFWTGU. 5QOG YQTMGTU WVKNK\G TGURKTCVQT[ FGHKEKGPE[ (RGVKVGU) HQT EQWPVGT UGNGEVKPI J[DTKFU. 6JKU QRGTCVGU D[ WUKPI QPG RTQVQVTQRJKE OCVKPI V[RG UVTCKP ECTT[KPI C TGURKTCVQT[ OWVCVKQP CPF ETQUUKPI VJKU YKVJ CPQVJGT OCVKPI V[RG VJCV KU TGURKTCVQT[RTQHKEKGPVÿ DWV YJKEJ ECTTKGU CP CWZQVTQRJ[ (FGTKXGF URQPVCPGQWUN[ QT XKC OWVCIGPGUKU). +P TCTG OCVKPI (TGHGTTGF VQ CDQXG)ÿ OCVKPI V[RG CWZQVTQRJU CTG ETQUUGF YKVJ RGVKVG RTQVQVTQRJKE KPFWUVTKCN UVTCKPU. +P GKVJGT ECUGÿ QPN[ J[DTKFU VJCV CTG TGURKTCVQT[-RTQHKEKGPV CPF RTQVQVTQRJKE CTG CDNG VQ TGEQXGT QP UGNGEVKQP OGFKWO (*COOQPF 199õ). òKPCNN[ÿ C HNQY E[VQOGVTKE DCUGF OGVJQF JCU DGGP FGXGNQRGF VJCV CNNQYU J[DTKFU VQ DG KFGPVKHKGF D[ WUKPI VYQ-EQNQWT HNQY E[VQOGVTKE EGNN UQTVKPI. +P VJKU RTQEGFWTGÿ QPG RCTGPVCN UVTCKP KU NCDGNGF YKVJ C ITGGP HNWQTGUEGPV EGNN VTCEMKPI F[Gÿ CPF VJG QVJGT KU NCDGNGF YKVJ CP QTCPIG HNWQTGUEGPV EGNN VTCEMKPI F[G. óJGP OKZGF VQIGVJGT WPFGT EQPFKVKQPU VJCV CNNQY OCVKPIÿ J[DTKFU QEEWTTKPI CV HTGSWGPEKGU QH CTQWPF 1 RGT OKNNKQP ECP DG KFGPVKHKGF CPF UQTVGF QP VJG DCUKU QH VJGKT FWCN QTCPIG CPF ITGGP HNWQTGUEGPEG (òKI.3þ $GNN GV CN. 199÷). 6JG CDKNKV[ QH HNQY E[VQOGVT[ VQ GZCOKPG NCTIG PWODGTU QH EGNNU CV TCVGU QH 1000'U RGT UGEQPF CNNQYU VJKU OGVJQF VQ QXGTEQOG RTQDNGOU CUUQEKCVGF YKVJ NQY QT TCTG OCVKPI GHHKEKGPEKGUÿ YKVJQWV VJG PGGF VQ KPVTQFWEG UGNGEVCDNG IGPGVKE OCTMGTU. 2.3.3 /WVCIGPGUKU +PVTQFWEVKQP QH JKIJN[ URGEKHKE OWVCVKQPU ECP DG CEJKGXGF D[ Tøý# UVTCVGIKGU. 6JGUG OC[ DG FGUKTCDNG YJGTG VJG IGPG QH KPVGTGUV CPF PCVWTG QH OWVCVKQP KU HWNN[ FGHKPGF. *QYGXGTÿ VJG CRRNKECVKQP QH Tøý# KU NKOKVGF HQT TGCUQPU FKUEWUUGF KP 5GEVKQP 3.1. %NCUUKECN OWVCIGPGUKU YKNN QHVGP NGCF VQ C JKIJ TCVGU QH KPEQTRQTCVKQP QH FGNGVGTKQWU IGPG EJCPIGU CU YGNN CU KORTQXGOGPVU UWEJ VJCV QEEWTTGPEG QH EGNN V[RGU YKVJ QPN[ VJG KORTQXGF IGPG HWPEVKQP KU WUWCNN[ XGT[ TCTG. /QTGQXGTÿ VJG EQORNGZ IGPQOKEU (FKRNQKF[ CPF RQN[RNQKFKGU) QH KPFWUVTKCN [GCUVU OGCP VJCV OQTG VJCP QPG IGPG EQR[ OC[ PGGF VQ DG OWVCIGPK\GF KP QTFGT VQ UGG FGUKTGF CNVGTCVKQPU KP RJGPQV[RGU. *GPEGÿ WPNGUU VJG[ CTG FQOKPCPVÿ OWVCVKQPU CTG PQV GCUKN[ FGVGEVGF KP FKRNQKFU QT RQN[RNQKFU. $GECWUG QH VJG PWOGTKECN TGUVTKEVKQPU KP CEJKGXKPI FGUKTGF OWVCVKQPU KP OQTG VJCP QPG EQR[ QH C VCTIGVGF IGPG OQUV YQTMGTU UVTGUU C RTGHGTGPEG VQ WUG JCRNQKF UVTCKPU KP OWVCIGPGUKU RTQITCOOGU (,QJPUVQP CPF úDGTOCP 19û9þ 2TGVQTKWU ù000). *QYGXGTÿ FGURKVG VJG PWOGTKECN EJCNNGPIGÿ KV JCU RTQXGP RQUUKDNG VQ QDVCKP WUGHWN OWVCPVU HTQO RQN[RNQKF UVTCKPU. òQT GZCORNGÿ OWVCIGPGUKU JCU DGGP CRRNKGF VQ RQN[RNQKF S. cerevisiae CPF S. carlsbergensis DTGYKPI UVTCKPU (,QJPUVQP CPF úDGTOCP 19û9).
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ôTGGP òNWQTGUEGPEG (KI. 3. 7UG QH HNWQTGUEGPEG CPF (#%5 VQ KUQNCVG OCVGF J[DTKFU YKVJQWV VJG PGGF HQT IGPGVKE OCTMGTU. %GNNU QH QRRQUKVG OCVKPI V[RG YGTG UVCKPGF YKVJ F[GU VJCV GOKV FKHHGTGPV HNWQTGUEGPV EQNQWTU (ITGGP QT QTCPIG). óJGP EGNNU OCVGF VJG[ DGECOG FWCN-UVCKPGF CPF YGTG FKUVKPIWKUJCDNG HTQO QTKIKPCN EGNNU. #) 2WTG UVTCKP 1 UVCKPGF YKVJ C ITGGP HNWQTGUEGPV EGNN VTCEMGT F[Gþ $) RWTG UVTCKP ù UVCKPGF YKVJ CP QTCPIG HNWQTGUEGPV EGNN VTCEMGT F[Gþ %) UVTCKPU 1 CPF ù KOOGFKCVGN[ CHVGT OKZKPIþ ø) RQRWNCVKQP % CHVGT QXGTPKIJV KPEWDCVKQP VQ CNNQY OCVKPI. ò#%5 QH VJG FWCN UVCKPGF EGNNU YKVJKP VJG RQN[IQPÿ CNNQYGF HQT KUQNCVKQP QH J[DTKFU CPF UWDUGSWGPV EQPHKTOCVKQP QH IGPQV[RGö RJGPQV[RG (UGG $GNN GV CN. 199÷).
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ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
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2.3.4 ÿTQVQRNCUV þ URJGTQRNCUV HWUKQP 2TQVQRNCUV QT URJGTQRNCUV HWUKQPU QHHGT C OGCPU QH IGPGVKE VTCPUHGT YJGTG KPVGTURGEKHKE DTGGFKPI KU PQV RQUUKDNGÿ QT YJGTG GZEJCPIGU DGVYGGP FKHHGTGPV URGEKGU CPF IGPGTC CTG FGUKTGF (4WUUGNN CPF 5VGYCTV 19û9þ 2TGVQTKWU ù000). òWUKQP ECP CNUQ DG WUGF VQ KPETGCUG VJG RNQKF[ QH UVTCKPUÿ G.I. HWUKQP QH C JCRNQKF YKVJ FKRNQKF VQ ETGCVG VTKRNQKF EGNNU. 6JG RTQEGUU KPXQNXGU TGOQXCN QH EGNN YCNNU HTQO [GCUV EGNNU D[ VTGCVOGPV YKVJ N[VKE GP\[OGU KP QUOQVKECNN[ UWRRQTVKXG OGFKWOÿ OKZKPI QH URJGTQRNCUVU HTQO VYQ FKUVKPEV UVTCKPU KP VJG RTGUGPEG QH C HWUQIGPKE CIGPV UWEJ CU RQN[GVJ[NGPG IN[EQNÿ FKOGVJ[N UWNRJQZKFG CPF %Cëë HQNNQYGF D[ C RGTKQF QH KPEWDCVKQP VQ CNNQY HWUKQP VQ QEEWT. 6JG HWUGF EGNNU CTG CNNQYGF VQ TGIGPGTCVG VJGKT EGNN YCNNU KP CP QUOQVKECNN[ UVCDKNK\GF CICT OGFKWO. ýWENGCT HWUKQP HQNNQYU CPF FGUKTCDNG CPF WPFGUKTCDNG VTCKVU QH RCTGPVCN UVTCKPU ECP TGEQODKPG KP QHHURTKPI. 6JG RTQFWEVU QH HWUKQPU CTG FKHHKEWNV VQ RTGFKEV DGECWUG VJGTG KU PQV CP CXGTCIKPI QH VJG EJCTCEVGTKUVKEU QH VJG RCTGPVCN UVTCKPU. 4CVJGT VJGTG VGPFU VQ DG C EJTQOQUQOCN FQOKPCPEG D[ VJG PWENGWU QH QPG FQPQT YKVJ NQUU QH OQUV QH VJG PWENGCT IGPQOG QH VJG UWDQTFKPCVG PWENGWU. 6JG OGEJCPKUO QH HQTOCVKQP QH J[DTKFU KU PQV HWNN[ WPFGTUVQQF DWV RTQDCDN[ EQPUKUVU QH VJG VTCPUHGT QH QPN[ C HGY IGPGU QT OC[DG C YJQNG EJTQOQUQOG HTQO QPG PWENGWU VQ CPQVJGT KP VJG HWUGF EGNN (5RGPEGT GV CN. 1990). 6JG PCVWTG QH KPVGTURGEKHKE CPF KPVGTIGPGTKE J[DTKFU CTG EQPUKUVGPV YKVJ VJG UKPING-EJTQOQUQOG VTCPUHGT EQPEGRV (øWVEJGT 19÷1þ 5RGPEGT GV CN. 1990). 6JG KPKVKCN PWENGCT HWUKQPU HQTOGF KP VJG RTQEGUU VGPF VQ DG OKVQVKECNN[ WPUVCDNG CU VJG RTQEGUU QH UWDQTFKPCVG EJTQOQUQOCN NQUU QEEWTU. 6JKU KPUVCDKNKV[ KU KPETGCUKPIN[ RTQOKPGPV VJG OQTG FKUVCPVN[ TGNCVGF VJG RCTGPVCN [GCUVU (/QTICP 19÷3). 6JGTG KU VJGTGHQTG C PGGF VQ UWDEWNVWTG J[DTKFU VJTQWIJ OCP[ IGPGTCVKQPU KP QTFGT VQ QDVCKP UVCDNG UVTCKPU. # OCjQT VGEJPKECN TGSWKTGOGPV KU VQ QRVKOK\G VJG GHHKEKGPE[ QH VJG HWUKQP RTQEGUU UQ VJCV JKIJGT PWODGTU QH J[DTKFU CTG HQTOGFÿ IKXKPI ITGCVGT RQVGPVKCN VQ CEJKGXG VJG FGUKTGF IGPGVKE EQODKPCVKQPU. +P VJKU TGICTFÿ GNGEVTQHWUKQP VGEJPKSWGU JQNF RTQOKUG (5EJPGVVNGT GV CN. 19÷í). ôGPGVKE CPCN[UGU QH HWUKQP RTQFWEVU TGXGCN C TCPIG QH RTQFWEVU. òQT GZCORNGÿ KP GZRGTKOGPVU KPXQNXKPI HWUKQP DGVYGGP JCRNQKF S. cerevisiae CPF S. diastaticusÿ VJTGG FKUVKPEV V[RGU QH J[DTKFU YGTG RTQFWEGF. 6JG HKTUV V[RG TGRTGUGPVGF CDQWV ÷0ð QH RTQFWEVU CPF ECTTKGF VJG S. diastaticus PWENGWU YKVJ VJG TGURKTCVQT[HWPEVKQPCN OKVQEJQPFTKC QH VJG S. cerevisiae JCRNQKF. 6JG UGEQPF V[RG QEEWTTGF CV CDQWV 1ûð HTGSWGPE[ YCU C FKRNQKF J[DTKF QH DQVJ RCTGPVUÿ CPF VJG VJKTF V[RG YGTG VTKRNQKF J[DTKFU HQTOGF D[ HWUKQP DGVYGGP VYQ S. cerevisiae URJGTQRNCUVU CPF QPG S. diastaticus URJGTQRNCUV (UGG ©XCPU 1990). +P OQUV ECUGUÿ GURGEKCNN[ YJGTG OQTG FKXGTIGPV UVTCKPU V[RGU CTG HWUGFÿ J[DTKFU CTG RQN[RNQKF YKVJ OCTMGF CPGWRNQKFKGU. 6JG OQUV NKMGN[ CRRNKECVKQP QH VJG RTQVQRNCUVöURJGTQRNCUV HWUKQP VGEJPQNQI[ KU KP VJG DTGYKPI KPFWUVT[ YJGTG VJG EQORNGZ IGPGVKE OCMGWR QH UVTCKPU TGPFGTU VJGO NCTIGN[ TGHTCEVQT[ VQ UVCPFCTF OCVKPI RTQEGFWTGU (UGG %JCRVGT ñ). #NVJQWIJ GCTN[ CVVGORVU CV HWUKPI DTGYGT'U [GCUVU NGF VQ HQTOCVKQP QH KPHGTKQT J[DTKFU VJCV RTQFWEGF QHH-HNCXQWTUÿ KPFWUVTKCN UVTCKP KORTQXGOGPVU JCXG DGGP CEJKGXGF XKC URJGTQRNCUV HWUKQP (6TKXGFK GV CN. 19÷õþ ©XCPU 1990þ 5RGPEGT GV CN. 1990þ 4GGF CPF ýCIQFCYKVJCPC 1991).
3÷
2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
2.3.5 %[VQFWEVKQP %[VQFWEVKQP KPXQNXGU VJG WUG QH QPG OCVKPI V[RG ECTT[KPI C kar OWVCVKQP VJCV KPJKDKVU PWENGCT HWUKQP (MCT[QICO[) CHVGT OCVKPI. 6JG kar OWVCPVU CTG OCVGF YKVJ UVTCKPU QH QRRQUKVG OCVKPI V[RG CPF [KGNF J[DTKFU VJCV TGVCKP VJG PWENGCT EQORQUKVKQP QH QPG RCTGPV DWV JCXG C OKZGF E[VQRNCUOKE OCMGWR (,QJPUVQP 1990). 6JG EQPEGRV VJCV UQOG E[VQFWEVCPVU OC[ ECTT[ ET[RVKE QT RJGPQV[RKECNN[ UKNGPV PWENGK HTQO VJG UGEQPF RCTGPV YCU KPVTQFWEGF D[ ýGX\INKCFQXC GV CN. (199÷ÿ ù00ù). 6JG RTQDNGO QH PQP-OCVKPI UVTCKPU ECP DG CFFTGUUGF VJTQWIJ E[VQFWEVKQP. òQT GZCORNGÿ +PIG-8GEJVQOQX GV CN. (19÷õ) UJQYGF VJCV J[DTKFK\CVKQP QH Saccharomyces [GCUVU YKVJ VJG UCOG OCVKPI V[RG EQWNF DG CEJKGXGF XKC E[VQFWEVKQPÿ GKVJGT URQPVCPGQWUN[ QT CHVGT 78-KTTCFKCVKQP. %[VQFWEVKQP ECP CNUQ TGUWNV KP UGNHFKRNQKFK\CVKQP KP UVTCKPU QH Saccharomyces FWG VQ HWUKQP QH UKDNKPI PWENGK KP VJG JGVGTQMCT[QVKE \[IQVG (2QNCKPC GV CN. 1993). %[VQFWEVKQP ECP DG WUGF VQ VTCPUHGT UWRGTKQT OKVQEJQPFTKCN HWPEVKQP CPF [GCUV GZVTCEJTQOQUQOCN HCEVQTU UWEJ CU RNCUOKFU CPF MKNNGT HCEVQTU (,QJPUVQP 1990þ 2TGVQTKWU ù000). +P HGTOGPVCVKQP VGUVU YKVJ DTGYGT'U [GCUV E[VQFWEVCPVUÿ KV YCU FGVGTOKPGF VJCV OKVQEJQPFTKC CPF RGTJCRU GZVTCOKVQEJQPFTKCN HCEVQTU ECP CHHGEV HGTOGPVCVKQP TCVGUÿ GVJCPQN VQNGTCPEGÿ HNQEEWNCVKQP CPF HNCXQWT EQORQWPFU (,QJPUVQP 1990). +PVTQFWEVKQP QH VJG FU4ý# XKTWU-NKMG MKNNGT HCEVQTU KPVQ [GCUV UVTCKPU ECP DG CP GHHGEVKXG CRRTQCEJ VQ FGCNKPI YKVJ 'YKNF [GCUV' EQPVCOKPCVKQP QH HGTOGPVCVKQPU. ;QWPI (19÷3) UJQYGF VJCV MKNNGT HCEVQT ECP DG KPVTQFWEGF VQ DTGYKPI UVTCKPU YKVJQWV CP[ FGNGVGTKQWU GHHGEV QP DGGT SWCNKV[. 5KOKNCTN[ÿ E[VQFWEVKQP VGEJPKSWGU JCXG DGGP KORQTVCPV KP FGXGNQRKPI UCMG [GCUVU YKVJ GZEGNNGPV HGTOGPVCVKXG EJCTCEVGTKUVKEU RNWU VJG CDKNKV[ VQ 'MKNN' YKNF [GCUVU (úPKUJK 1990). 2.3.6 SETGGPKPI UVTCKPU HQT KORTQXGOGPVU øGTKXCVKQP QH PQXGN J[DTKFUÿ PGY OWVCPVUÿ KPVGTIGPGTKEöKPVGTURGEKHKE UVTCKPU QT E[VQFWEVCPVU KU QPN[ RCTV QH VJG RTQEGUU QH UVTCKP KORTQXGOGPV. #HVGT VJG IGPGVKE RTQEGUU(GU) UGXGTCN NGXGNU QH UETGGPKPIÿ UGNGEVKQP CPF EJCTCEVGTK\CVKQP CTG PGEGUUCT[. +P IGPGTCN VGTOUÿ UVTCKPU PGGF VQ DG GXCNWCVGF HQT ITQYVJ TCVGUÿ [KGNFU QP FKHHGTGPV UWDUVTCVGUÿ VQNGTCPEG VQ GPXKTQPOGPVCN EJCNNGPIGUÿ CPF QWVRWV QH URGEKHKE RTQFWEVU (G.I. GP\[OGUÿ XKVCOKPUÿ 4ý#öPWENGQVKFGUÿ COKPQ CEKFU UWEJ CU INWVCOCVGÿ QTICPKE CEKFUÿ JGVGTQNQIQWU RTQVGKPU). +P OQTG URGEKHKE VGTOUÿ YKPGÿ FKUVKNNGT'U CPF DTGYGT'U [GCUVU CTG GXCNWCVGF HQT HGTOGPVCVKXG CEVKXKV[ÿ HNCXQWT CPF GVJCPQN VQNGTCPEGÿ YJGTGCU DCMGT'U [GCUV YKNN DG VGUVGF HQT CGTQDKE ITQYVJ [KGNFÿ URGGF QH HGTOGPVCVKQP KP FKHHGTGPV FQWIJU CPF DTGCF OCMKPI RTQEGUUGUÿ CDKNKV[ VQ HGTOGPV KP GZVTGOGN[ JKIJ UWICT EQPEGPVTCVKQPUÿ CPF RQUUKDN[ VQNGTCPEG VQ HTQ\GP FQWIJ OGVJQFU. óKPG CPF DCMGT'U [GCUVU YQWNF CNUQ DG UETGGPGF HQT VQNGTCPEG VQ FT[KPI CPF TG-J[FTCVKQPÿ YJGTG PGY UVTCKPU QH 'CEVKXG' QT 'KPUVCPV' FT[ [GCUV CTG UQWIJV. 6JG GZCEV FGVCKNU QH JQY VJGUG RTQRGTVKGU CTG GXCNWCVGF HQT KPFKXKFWCN UVTCKPU KU FGRGPFGPV WRQP VJG URGEKHKE KPFWUVTKCN CRRNKECVKQP CPF GXGP WRQP KPFKXKFWCN EQOOGTEKCN RTQFWEVKQP RNCPVU CPF RTQEGUUGU VJCV PGY UVTCKPU CTG VCTIGVGF VQYCTFU. *GPEGÿ KV KU WR VQ VJG KPFKXKFWCN TGUGCTEJGT VQ FGXGNQR ETKVKECN NCDQTCVQT[
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
39
VGUVU VJCV DGUV OKOKE VJG CRRNKECVKQPU QH [GCUVU KP KPFWUVTKCN UKVWCVKQPU. +P VJKU TGICTFÿ KV KU XKTVWCNN[ RQKPVNGUU DGIKPPKPI VJG IGPGVKE CRRTQCEJ WPNGUU C OGCPKPIHWN UETGGP CPF QVJGT VGUVU JCXG DGGP FGXGNQRGF VJCV VTWN[ GXCNWCVG VJG ECRCDKNKV[ QH UVTCKPU HQT FGUKTGF CRRNKECVKQPU.
2.4 +ORTQXGOGPVU VQ UVTCKPU XKC ENCUUKECN IGPGVKEU øGURKVG VJG VGEJPKECN EJCNNGPIGUÿ ENCUUKECN IGPGVKE UVTCVGIKGU JCXG DGGP WUGF VQ RTQFWEG [GCUVU VJCV GZJKDKV KORTQXGF RGTHQTOCPEG YKVJKP VJG DCMKPIÿ YKPGÿ FKUVKNNKPIÿ DTGYKPIÿ KPFWUVTKCNöHWGN CNEQJQN CPF HQQF [GCUV KPFWUVTKGU (,QJPUVQP CPF úDGTOCP 19û9ÿ ©XCPU 1990þ ,CEQDUQP CPF 6TKXGFK 1990þ ,QJPUVQP 1990þ úFC CPF úWEJK 1990þ 4GGF CPF ýCIQFCYKVJCPC 1991þ $CTTG GV CN. 1993þ ©jKQHQT GV CN. 199íþ *CPUGP CPF îKGNNCPF-$TCPFV 199õþ 4CPFG\-ôKN GV CN. 1999þ øGSWKP ù001). 5QOG GZCORNGU QH UVTCKP KORTQXGOGPV D[ ENCUUKECN IGPGVKEU CTG IKXGP KP 6CDNG 3. +P QTFGT VQ FGOQPUVTCVG VJG RTQITGUU VJCV ECP DG CEJKGXGF YKVJ ENCUUKECN IGPGVKE CRRTQCEJGU YG JKIJNKIJV GZCORNGU QH KORTQXGOGPVU KP jWUV QPG KPFWUVTKCN CRRNKECVKQP QH [GCUVUÿ PCOGN[ DCMKPI. #PIGNQX GV CN. (199õ) WUGF EJGOKECN OWVCIGPGUKU VQ IGPGTCVG KORTQXGF OCNVQUG-HGTOGPVKPI [GCUVU. /CVKPI CPF UETGGPKPI UVTCVGIKGU YGTG WUGF VQ IGPGTCVG PGY UVTCKPU VJCV EQODKPG JKIJ OCNVQUG-WVKNK\KPI CEVKXKV[ TGSWKTGF HQT RNCKP FQWIJ NGCXGPKPIÿ YKVJ NQY KPXGTVCUG CEVKXKV[ CUUQEKCVGF YKVJ GHHKEKGPV JKIJ UWICT FQWIJ HGTOGPVCVKQP (LQvG\ GV CN. 199ù). +P CP CNVGTPCVKXG CRRTQCEJ VQ CEJKGXKPI VJG IQCN QH DTQCF FQWIJ TCPIG [GCUVUÿ ENCUUKECN DTGGFKPI CPF UGNGEVKQP RTGUUWTGU YGTG WUGF VQ KUQNCVG RQQNU QH JCRNQKF [GCUV UVTCKPU YKVJ UVTQPI RNCKP FQWIJ QT JKIJ UWICT FQWIJ HGTOGPVKPI ECRCEKVKGU. 6JGUG YGTG VJGP OCVGF GP OCUUG VQ IGPGTCVG C DTQCF FQWIJ TCPIG UVTCKP VJCV HGTOGPVU GHHKEKGPVN[ KP RNCKP CPF JKIJ UWICT FQWIJU KP URKVG QH VJG HCEV VJCV KV JCU C JKIJ KPXGTVCUG CEVKXKV[ (*KIIKPU GV CN. ù001). /WVCPV UGNGEVKQP RTQITCOOGU JCXG CNUQ NGF VQ FGXGNQROGPV QH KPFWUVTKCN DCMGT•U [GCUV UVTCKPU VJCV CTG UWDUVCPVKCNN[ KPCEVKXG CV TGHTKIGTCVKQP VGORGTCVWTGUÿ DWV YJKEJ TGEQXGT HGTOGPVCVKXG CEVKXKV[ VQ PQTOCN NGXGNU CV VGORGTCVWTGU CDQXG 1íQ% (*QVVKPIGT GV CN. 199÷þ ô[UNGT CPF ýKGFGTDGTIGT ù00ù). 5WEJ UVTCKPU CTG RQVGPVKCNN[ WUGHWN HQT RTQNQPIGF UVQTCIG CU C EJKNNGF UWURGPUKQP QT CU TGHTKIGTCVGF EQORTGUUGF DNQEMU. # HWTVJGT GZCORNG QH OWVCPV KUQNCVKQP KU VJG QDVCKPKPI QH DCMGT•U [GCUVU VJCV UJQY GPJCPEGF UVTGUU TGUKUVCPEG FWTKPI KPKVKCVKQP QH HGTOGPVCVKQP (XCP øKjEM GV CN. ù000þ 6GWPKUUGP GV CN. ù00ù). +P PQTOCN UVTCKPU QH [GCUVU VJG QPUGV QH HGTOGPVCVKQP NGCFU VQ C UJWV FQYP QH UVTGUU TGUKUVCPEG HCEVQTU CU EGNNU IGCT WR HQT ITQYVJ-CUUQEKCVGF CEVKXKVKGU. *QYGXGTÿ VJG UQ-ECNNGF fil OWVCPVUÿ FGHKEKGPV KP fGTOGPVCVKQP iPFWEGF lQUU QH UVTGUU TGUKUVCPEGÿ QHHGT RQVGPVKCN CFXCPVCIGU KP HTQ\GP FQWIJ CRRNKECVKQPU YJGTG RTGNKOKPCT[ HGTOGPVCVKQP QEEWTU CPF EGNNU ECP DGEQOG UVTGUU-UGPUKVKXG RTKQT VQ FQWIJU TGCEJKPI HTGG\KPI RQKPV. úVJGTU JCXG CNUQ FGUETKDGF XCTKQWU ENCUUKECN IGPGVKE UVTCVGIKGU VQ QDVCKP [GCUVU YKVJ KORTQXGF RQVGPVKCN HQT HTQ\GP FQWIJ CRRNKECVKQPU (*KPQ GV CN. 19÷ûþ *CJP CPF îCYCK 1990þ ýCMCICYC CPF úWEJK 199í).
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
6CDNG 3. 5QOG GZCORNGU QH KORTQXGOGPVU VQ KPFWUVTKCN [GCUV UVTCKPU XKC ENCUUKECN IGPGVKEU +PFWUVT[ $TGYKPI
6GEJPKSWG òWUKQP
%NCKOGF DGPGHKV ©PJCPEGF HNQEEWNCVKQP
$TGYKPI
/WVCPV UGNGEVKQP /WVCPV UGNGEVKQP /CVKPI òWUKQP /CVKPI /WVCIGPGUKU /CVKPI /WVCIGPGUKU
øGTGRTGUUGF OCNVQUG WRVCMG
4GHGTGPEG 7TCPQ GV CN. (1993C CPF D) 5VGYCTV GV CN. (19֖)
+ORTQXGF GVJCPQN VQNGTCPEG
îQTJQNC (19÷3)
+ORTQXGF CVVGPWCVKQPÿ +PVTQFWEGF OGNKDKCUG CEVKXKV[ +PVTQFWEGF OGNKDKCUG +ORTQXGF HTQ\GP FQWIJ CEVKXKV[ $TQCF TCPIG FQWIJ HGTOGPVCVKQP +ORTQXGF PKVTQIGP CUUKOKNCVKQPÿ CPF HGTOGPVCVKQP MKPGVKEU #NEQJQN VQNGTCPEG LQY UWNRJKFG RTQFWEVKQP +PVTQFWEGF MKNNGT HCEVQT K.lactis Z S. cerevisiae HWUCPVU +PVTQFWEGF CO[NQN[VKE CEVKXKV[ +PETGCUGF RNQKF[ CPF HNQEEWNCVKQP 4CKUGF NGXGNU QH RTQVGKP CPF UWNRJWT COKPQ CEKFU
,QJPUVQP (1990) ,QJPUVQP (1990) 8KPEGPV GV CN. (1999) 8CP øKjEM GV CN.(ù000) *KIIKPU GV CN. (ù001) 5CNOQP CPF $CTTG (199÷) 7PCNFK GV CN. (ù00ù) 4QOCPQ GV CN. (19÷ñ) úWEJK GV CN. (19÷3) òCTCJPCM GV CN. (19÷õ) 5KNNU GV CN. (19÷3) ,CXCFGMCT GV CN. (199ñ) ,QJPUVQP CPF úDGTOCP (19û9)
øKUVKNNKPI øKUVKNNKPI øKUVKNNKPI $CMKPI $CMKPI $CMKPI óKPG óKPG óKPG 5CMG #NEQJQN #NEQJQN #NEQJQN òQQF [GCUVU
/WVCIGPGUKU /CVKPI %[VQFWEVKQP òWUKQP 4CTG OCVKPI òWUKQP /WVCIGPGUKU
2.4.1 +ORTQXGF OCNVQUG WVKNK\CVKQP KP KPFWUVTKCN S. cerevisiae: C WPKQP QH HWPEVKQPCN CPCN[UKU CPF ENCUUKECN IGPGVKEU KP UVTCKP KORTQXGOGPV 6JG ½-INWEQUKFGUÿ YJKEJ KPENWFGU OCNVQUG CPF OCNVQVTKQUGÿ CTG VJG OQUV CDWPFCPV HGTOGPVCDNG UWICTU KP KPFWUVTKCN CRRNKECVKQPU. /CNVQUG OGVCDQNKUO KU C RCTVKEWNCTN[ ETWEKCN RCTCOGVGT KP VJTGG QH VJG NCTIGUV KPFWUVTKCN CRRNKECVKQPU QH [GCUVUÿ RNC[KPI C OCjQT TQNG KP HGTOGPVCVKQP QH FQWIJUÿ DTGYGT'U YQTVUÿ CPF FKUVKNNGT'U OCUJGU (5VCODWM CPF FG #TCWjQ ù001). 2.4.1.1 Genomic localization of /#L loci ôGPGVKE CPCN[UKU QH NCDQTCVQT[ UVTCKPU QH S. cerevisiae UJQYU WR VQ HKXG MAL NQEK (MAL1ÿ MAL2ÿ MAL3, MAL4 CPF MAL6). 6JG RTGUGPEG QH CP[ QPG QH VJGUG GPCDNGU C [GCUV VQ HGTOGPV OCNVQUG. +P YKNF V[RG NCDQTCVQT[ UVTCKPU QH S. cerevisiae OCNVQUG OGVCDQNKUO KU KPFWEGF D[ OCNVQUGÿ TGRTGUUGF D[ INWEQUGÿ CPF CV C DCUCN NGXGN KP VJG CDUGPEG QH DQVJ VJGUG UWICTU. ôGPGVKE OCRRKPI CUUKIPGF GCEJ MAL NQEWU VQ UGRCTCVG EJTQOQUQOGU CU HQNNQYU: MAL1 EJTQOQUQOG 8++þ MAL2 EJTQOQUQOG +++þ MAL3 EJTQOQUQOG ++þ MAL4 EJTQOQUQOG ï+: CPF MAL6 EJTQOQUQOG 8++ (/QTVKOGT GV CN. 199ù). 7UKPI RTQDGU FGTKXGF HTQO VJG MAL6 NQEWUÿ TGUVTKEVKQP HTCIOGPV NGPIVJ RQN[OQTRJKUOU JCXG DGGP KFGPVKHKGF HQT GCEJ QH VJG HWPEVKQPCN MAL NQEK KFGPVKHKGF KP NCDQTCVQT[ UVTCKPU CNNQYKPI C YKFG TCPIG QH UVTCKPU VQ DG GZ-
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
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COKPGF (%JQY GV CN. 19÷3). #NN UVTCKPU GZCOKPGF EQPVCKPGF C MAL1 NQEWU QT C PQP-HWPEVKQPCN CNNGNG KPFKECVKPI VJCV VJG MAL1 NQEWU OC[ JCXG DGGP C RTQIGPKVQT HQT CNN QH VJG QVJGT MAL NQEK (%JQY GV CN. 19÷3). òWTVJGTOQTGÿ UVWFKGU QP VJG UVCPFCTF MAL NCDQTCVQT[ UVTCKPU FGVGTOKPGF VJCV CNVJQWIJ QPN[ QPG HWNN[ HWPEVKQPCN NQEWU YCU RTGUGPV KP VJG UVTCKPUÿ GCEJ UVTCKP EQPVCKPGF QVJGT IGPQOKE UGSWGPEGU YKVJ JQOQNQI[ VQ VJG OCNVCUG IGPG. 6JGUG KPENWFGF ET[RVKE MAL NQEK (K.G. PQPHWPEVKQPCN QT QPN[ RCTVKCNN[ HWPEVKQPCN MAL IGPGU) CPF UGSWGPEGU YKVJ UKIPKHKECPV JQOQNQI[ VQ VJG OCNVCUG RTQDGÿ DWV NCEMKPI KP MAL HWPEVKQPU (/KEJGNU CPF ýGGFNGOCP 19÷í). #PCN[UKU QH VJTGG RCTVKCNN[ HWPEVKQPCN MAL1 CNNGNGU KPFKECVGF VJCV VJG[ YGTG FGTKXGF HTQO VJG MAL1 HWPEVKQPCN NQEWU D[ C XCTKGV[ QH FGNGVKQPU QT TGCTTCPIGOGPVU QH EJTQOQUQOCN UGSWGPEGU (%JCTTQP CPF /KEJGNU 19÷÷). úPG QH VJGUG RCTVKCNN[ HWPEVKQPCN MAL1 CNNGNGU EQPVCKPGF C OCNVQUG RGTOGCUG YKVJ NQYGTGF UGSWGPEG JQOQNQI[ VQ VJG MAL1 OCNVQUG RGTOGCUG (%JCTTQP CPF /KEJGNU 19÷÷). 2.4.1.2 Structure of the /#L loci 6JG MAL6 NQEWU JCU DGGP WUGF CU C OQFGN HQT VJG WPFGTUVCPFKPI QH VJG UVTWEVWTG CPF HWPEVKQP QH CNN QH VJG XCTKQWU MAL NQEK (ýGGFNGOCP 1991). 6JG VJTGG IGPGU QH VJG MAL6 NQEWU CTG NQECVGF YKVJKP C TGIKQP QH CRRTQZKOCVGN[ û MD CPF VJG QTFGT QH VJG IGPGU KU MAL6R (TGIWNCVQT[ RTQVGKP)ÿ MAL6T (OCNVQUG VTCPURQTVGT) CPF MAL6S (OCNVCUG)ÿ YKVJ MAL6S DGKPI NQECVGF ENQUGUV VQ VJG VGNQOGTG (ýGGFNGOCP 1991). 6JG OCNVQUG VTCPURQTVGT JCU CNUQ DGGP UJQYP VQ DG TGURQPUKDNG HQT WRVCMG QH OCNVQVTKQUG (øC[ GV CN. ù00ù). 6JG UGSWGPEKPI QH VJG IGPQOG QH VJG S. cerevisiae NCDQTCVQT[ UVTCKP 5ù÷÷E JCU EQPHKTOGF C JKIJ FGITGG QH DQVJ UVTWEVWTCN CPF UGSWGPEG JQOQNQI[ DGVYGGP VJG MAL1, MAL3 CPF MAL6 NQEK (8QNEMCGTV GV CN. 199û). +V VJGTGHQTG CRRGCTU VJCV VJG MAL6 NQEWU KU C WUGHWN RCTCFKIO HQT WPFGTUVCPFKPI VJG UVTWEVWTG CPF HWPEVKQP QH VJG QVJGT MAL NQEK. $G[QPF VJG DCUKE VJTGG IGPG UVTWEVWTGÿ UQOG XCTKCVKQP KP VJG UVTWEVWTGU QH VJG MAL6 CPF MAL3 NQEK QEEWT FWG VQ FWRNKECVKQP QH UGSWGPEGU ñ' QH VJG TGIWNCVQT[ IGPG (%JQY GV CN. 19÷9). +P VJG ECUG QH VJG MAL6 NQEWUÿ VJGTG KU C FWRNKECVKQP QH VJG MAL TGIWNCVQT[ IGPG WRUVTGCO QH VJG PQTOCN HWPEVKQPCN MAL TGIWNCVQTÿ YJKEJ UJCTGU GZVGPUKXG JQOQNQI[ YKVJ MAL6R. 6JG FWRNKECVGF TGIKQP CV VJG MAL3 NQEWU EQPUKUVU QH C RCKT QH CRRTQZKOCVGN[ 10 MD FKTGEV TGRGCVU EQPVCKPKPI CNN QH VJG MAL TGIWNCVQT[ NQEWU CPF RCTVU QH VJG OCNVQUG RGTOGCUG NQEWU (%JQY GV CN. 19÷9). 6JG MAL1 NQEWUÿ KFGPVKHKGF D[ UGSWGPEKPI QH VJG 5ù÷÷E IGPQOGÿ JCU CP ½-INWEQUKFCUG WRUVTGCO QH MAL1R VJCV KU PQV HQWPF KP VJG MAL6 NQEWU (8QNEMCGTV GV CN 199û). 6JG OCNVCUG IGPGU QH VJG MAL1 CPF MAL3 NQEK DQVJ UJCTG XGT[ JKIJ JQOQNQI[ YKVJ VJG MAL6 OCNVCUG (òGWGTOCPP GV CN. 199ñþ 8QNEMCGTV GV CN. 199û). #V VJG RTQVGKP NGXGNÿ C EQPUGPUWU UGSWGPEG ECP DG RTQFWEGFÿ HTQO YJKEJ GCEJ QH VJG VJTGG OCNVCUG RTQVGKPU QPN[ FKHHGTU D[ C UKPING COKPQ CEKF EJCPIG. 2.4.1.3 The /#L regulator 5GSWGPEKPI QH VJG MAL6R TGIWNCVQT JCU TGXGCNGF VJCV KV GPEQFGU C íû3 COKPQ CEKF RQN[RGRVKFG VJCV EQPVCKPU DQVJ C \KPE-EQQTFKPCVGF øý#-DKPFKPI OQVKH CPF RWVCVKXG
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
CEKFKE CEVKXCVQT FQOCKPU (5QNNKVVK CPF /CTOWT 19÷÷þ îKO CPF /KEJGNU 19÷÷). 6JG \KPE-EQQTFKPCVGF øý#-DKPFKPI FQOCKP JCU C %[Uõ OQVKHÿ YJKEJ KU JQOQNQIQWU VQ UGXGTCN QVJGT NQYGT GWMCT[QVKE CEVKXCVQTU ($CK CPF îQJNCY 1991). +P S. cerevisiaeÿ VJKU ITQWR KPENWFGU VJG LGW3ÿ ôCNíÿ 2RT1ÿ 7IC3ÿ #TI++ÿ *CR1ÿ 2FT1 CPF 2WV3 CEVKXCVQT RTQVGKPU ($CK CPF îQJNCY 1991). /CN4 DKPFU CU C FKOGT CPF ECP KFGPVKH[ VJG TGEQIPKVKQP OQVKH EöC ô% ý9 EöC ô EöI (5KTGPMQ CPF ýGGFNGOCP 199ñ). /CN4 ECP FKTGEVN[ UGPUG OCNVQUG KP VJG EGNN. 7PNKMG VJG GAL TGIWNQPÿ PQ TGRTGUUQT RTQVGKP GSWKXCNGPV VQ ôCN÷0 JCU DGGP HQWPFÿ DWV TCVJGTÿ CNN EQPUVKVWVKXG OWVCVKQPU QH MAL GZRTGUUKQP CTG NKPMGF VQ VJG MAL NQEWU (ôKDUQP GV CN. 199û). +P UGXGTCN MAL EQPUVKVWVKXG OWVCPVUÿ EQPUVKVWVKXG GZRTGUUKQP KU FGRGPFGPV WRQP OWVCVKQPU KP VJG %VGTOKPCN RQTVKQP QH VJG CEVKXCVQT RTQVGKP KPFKECVKPI VJCV VJKU TGIKQP FKTGEVN[ QT KPFKTGEVN[ UGPUGU OCNVQUG (ôKDUQP GV CN. 199û). /WVCPVU QH VJG MAL6 NQEWU JCXG DGGP RTQFWEGF VJCV EQPHGT EQPUVKVWVKXG GZRTGUUKQP QP MALT CPF MALS (6GP $GTIG GV CN. 19û3þ óCPI CPF ýGGFNGOCP 199õ). 6JGUG EQPUVKVWVKXG OWVCVKQPU JCXG DGGP OCRRGF VQ GKVJGT MAL6R QT C HQWTVJ IGPG CV VJG MAL NQEWU VGTOGF mal64. 6JG YKNF-V[RG MAL64 KU PQV PQTOCNN[ TGSWKTGF HQT OCNVQUG HGTOGPVCVKQPÿ FQGU PQV EQORNGOGPV OWVCVKQPU KP MAL6Rÿ CPF KU QPN[ RCTVKCNN[ JQOQNQIQWU VQ MAL6R (øWDKP GV CN. 19÷÷). óJKNUV VJG YKNF-V[RG MAL64 IGPG KU PQV KPXQNXGF KP OCNVQUG OGVCDQNKUOÿ VJG EQPUVKVWVKXG MAL64c OWVCPV JCU DGGP UJQYP VQ DG C INQDCN TGIWNCVQT[ IGPG CHHGEVKPI UGXGTCN FKHHGTGPV RCVJYC[U QH ½-INWEQUKFG OGVCDQNKUO (2GTMKPU CPF ýGGFNGOCP 19÷÷). 6YQ V[RGU QH EQPUVKVWVKXG OWVCVKQPU QH MAL6R JCXG DGGP KFGPVKHKGFþ VJQUG VJCV CTG EQPUVKVWVKXG DWV CTG UVKNN UWDjGEV VQ INWEQUG TGRTGUUKQPÿ CPF VJQUG VJCV CTG PQV UWDjGEV VQ INWEQUG TGRTGUUKQP (óCPI CPF ýGGFNGOCP 199õ). +P VJG OWVCPVU VJCV CTG UVKNN UWDjGEV VQ INWEQUG TGRTGUUKQPÿ VJG %-VGTOKPCN TGIKQP QH VJG RTQVGKP KU KORNKECVGF YJGTGCU KP VJG OWVCPVU VJCV CTG HWNN[ EQPUVKVWVKXGÿ OWVCVKQPU KP VJG %-VGTOKPCN FQOCKP CPF VJG NQUU QH C /KI1R-DKPFKPI UKVG WRUVTGCO QH VJG MAL6R UVTWEVWTCN IGPG CTG KORNKECVGF (óCPI CPF ýGGFNGOCP 199õ). %QORCTKUQP DGVYGGP VJG MPQYP YKNF-V[RG MAL TGIWNCVQT[ IGPGU JCU UJQYP VJCV CNN GZEGRV MAL4 CTG RJGPQV[RKECNN[ GSWKXCNGPV VQ VJG MAL6 NQEWU. 6JG GZEGRVKQPÿ VJG MAL4R CNNGNGÿ KU HQWPF KP UVTCKP 1í03-û#ÿ YJKEJ KU EQPUVKVWVKXG (%JCTTQP CPF /KEJGNU 19÷û) CPF YCU HQWPF CU C URQPVCPGQWU OWVCPV KP ï-TC[ OWVCIGPGUKU UVWFKGU (îJCP CPF ©CVQP 19û1). 6JGTG JCU DGGP PQ MAL4 NQEWU [GV KFGPVKHKGF VJCV KPFWEGU ÂYKNF-V[RG• TGIWNCVKQP QH MALT CPF MALS GZRTGUUKQP CPF VJWU KV KU RQUUKDNG VJCV VJG MAL4 NQEWU KVUGNH KU C RTQFWEV QH VJG OWVCIGPGUKU RTQEGFWTG. %QPUVKVWVKXG MAL OWVCVKQPU JCXG CNUQ DGGP TGRQTVGF HQT MAL1 (4QFKEKQ 19÷õ)ÿ CPF MAL2 (
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
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EQPUVKVWVKXG CEVKXCVQT VJCP YKVJ QVJGT TGIWNCVGF /CN4 RTQVGKPU (óCPI CPF ýGGFNGOCP 199õ). +V VJGTGHQTG CRRGCTU VJCV VJG /CNí4E EQPUVKVWVKXG CEVKXCVQT YCU VJG TGUWNV QH C TGEQODKPCVKQP GXGPV DGVYGGP C ET[RVKE MAL-JQOQNQIQWU UGSWGPEG CPF C MALR UGSWGPEG. 5KOKNCTN[ÿ UQOG QH VJG MAL2R EQPUVKVWVKXG IGPGU OC[ JCXG GXQNXGF D[ TGEQODKPCVKQP YKVJ C PQP-HWPEVKQPCN MAL3R (ôKDUQP GV CN. 199û). 2.4.1.4 Control of maltase and maltose permease 6JG FKXGTIGPVN[ CTTCPIGF MAL6T CPF MAL6S CTG UGRCTCVGF D[ ÷ûñ DR QH RTQOQVGT TGIKQP (*QPI CPF /CTOWT 19÷õ). 2TKOGT GZVGPUKQP UVWFKGU OCRRKPI VTCPUETKRV KPKVKCVKQP UKVGU KPFKECVGF VJCV 6#6# DQZGU CTG NQECVGF CV CRRTQRTKCVG RQUKVKQPU WRUVTGCO QH VJG DQVJ IGPGU (*QPI CPF /CTOWT 19÷õ). óKVJKP VJKU RTQOQVGT TGIKQP CTG VJG GNGOGPVU TGSWKTGF HQT VJG TGIWNCVGF GZRTGUUKQP QH VJG OCNVQUG RGTOGCUG CPF OCNVCUG IGPGUÿ KPENWFKPI /CN4 DKPFKPI UKVGU HQT KPFWEVKQP KP TGURQPUG VQ VJG RTGUGPEG QH OCNVQUGÿ CPF /KI1 DKPFKPI UKVGU HQT TGRTGUUKQP KP VJG RTGUGPEG QH INWEQUG. øGNGVKQP UVWFKGU JCXG CEEWTCVGN[ FGNKPGCVGF C UKPING TGIKQP QH VJG FKXGTIGPV RTQOQVGT VJCV RQUKVKXGN[ EQPVTQNU TGIWNCVKQP QH DQVJ VJG MALT CPF MALS IGPG XKC DKPFKPI YKVJ VJG /CN4 RTQVGKP. #NKIPOGPV QH VJG /KI1R EQPUGPUWU DKPFKPI UKVG CPF VJG MAL6 KPVGTIGPKE TGIKQP JCU KFGPVKHKGF HQWT UKVGU YKVJ UKIPKHKECPV JQOQNQI[ (*W GV CN. 199ñ). ôGN UJKHV CPCN[UKU JCU FGVGTOKPGF VJCV QH VJGUG HQWT UKVGUÿ VYQ DKPF /KI1 UVTQPIN[ÿ QPG YGCMN[ CPF VJG QVJGT WPFGVGEVCDNG (*W GV CN. 199ñ). 2TGXKQWU FGNGVKQP UVWFKGU VGPVCVKXGN[ KFGPVKHKGF QPG INWEQUG TGRTGUUKQP GNGOGPV YKVJKP VJG MAL RTQOQVGT (LGXKPG GV CN. 199ù) CPF VJKU YCU HQWPF VQ EQKPEKFG YKVJ QPG QH VJG UGSWGPEGU VJCV UVTQPIN[ DQWPF /KI1 (*W GV CN. 199ñ). øGNGVKQP QH GCEJ QH VJG /KI1 UKVGU JCU CNNQYGF VJG FGVGTOKPCVKQP QH VJG EQPVTKDWVKQP QH VJG UKVGU VQ GZRTGUUKQP QH DQVJ VJG MAL6T CPF MAL6S IGPGU. 6JG /KI1 UKVG RTQZKOCN VQ MAL6T EQPVTKDWVGU UKIPKHKECPVN[ VQ INWEQUG TGRTGUUKQP QH MAL6Tÿ YJKNG VJG /KI1 UKVG RTQZKOCN VQ VJG MAL6S IGPG EQPVTKDWVGU UKIPKHKECPVN[ VQ TGRTGUUKQP QH MAL6S. +P GCEJ ECUGÿ VJG FKUVCN UKVG JCU QPN[ C OQFGUV GHHGEV QP /KI1-FGRGPFGPV TGRTGUUKQPÿ KORN[KPI VJCV VJG TGRTGUUKQP UKVGU JCXG C TGNCVKXGN[ JKIJ FGITGG QH WPKFKTGEVKQPCN GHHGEVU YKVJKP VJG MAL FKXGTIGPV RTQOQVGT. 2.4.1.5 Genetic variation of MalR control 5GSWGPEG EQORCTKUQPU QH VJG MAL NQEK QH KPFWUVTKCN DCMGT'U [GCUVU KPFKECVG VJCV EQPUKFGTCDNG XCTKCVKQPU JCXG CTKUGP KP MALR - OQUV RTQDCDN[ CU C TGUWNV QH FQOGUVKECVKQP QH VJGUG [GCUVU CPF UGNGEVKQP HQT TCRKF OCNVQUG HGTOGPVCVKQP QXGT OCP[ IGPGTCVKQPU ($GNN GV CN. ù001). 6JG UGSWGPEG XCTKCVKQPU NGCF VQ INWEQUG KPUGPUKVKXKV[ CPF CDKNKV[ VQ GZRTGUU OCNVQUG RGTOGCUG CPF OCNVCUG CV JKIJ NGXGNU KP VJG CDUGPEG QH OCNVQUG (*KIIKPU GV CN. 1999C CPF Dþ øCP\K GV CN. ù000). +P C UGTKGU QH UVWFKGU QP C YKFGN[ WUGF KPFWUVTKCN [GCUV UVTCKP ECRCDNG QH CFCRVKPI HTQO INWEQUG HGTOGPVCVKQP VQ OCNVQUG HGTOGPVCVKQP TCRKFN[ÿ KV YCU HQWPF VJCV PKPG MAL NQEK YGTG RTGUGPV YKVJKP VJG VGVTCRNQKF UVTCKP. %NQPKPI QH UGXGTCN MALR CNNGNGU HTQO VJG UVTCKP KPFKECVGF VJG RTGUGPEG QH CV NGCUV VJTGG HWPEVKQPCN XCTKCPVU (*KIIKPU GV CN. 1999C CPF D). óJGP VJGUG IGPGU YGTG GZCOKPGF VQ FGVGTOKPG JQY VJG[ TGIWNCVGF GZRTGUUKQP QH VJG MALT CPF MALSÿ KV YCU HQWPF VJCV GCEJ IGPG FKURNC[GF FKHHGT-
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
GPV TGIWNCVQT[ EJCTCEVGTKUVKEU. úPG QH VJG IGPGU DGJCXGF KP C UKOKNCT YC[ VQ VJG MAL6 TGIWNCVQT[ IGPGÿ CPF FKTGEVGF VKIJVN[ TGIWNCVGF IGPG GZRTGUUKQP QH VJG OCNVCUG CPF OCNVQUG RGTOGCUG IGPGU. +P INWEQUGÿ GZRTGUUKQP YCU XGT[ NQYÿ YJKNUV QP OCNVQUG GZRTGUUKQP YCU JKIJ. 6JG QVJGT VYQ IGPGU EQPHGTTGF XCTKCDNG FGITGGU QH EQPUVKVWVKXKV[ CPF INWEQUG KPUGPUKVKXKV[ QP VJG GZRTGUUKQP QH VJG OCNVCUG CPF OCNVQUG RGTOGCUG IGPGU. #U C TGUWNV QH VJG RTGUGPEG QH VJKU UWKVG QH MAL TGIWNCVQTUÿ KV YCU HQWPF VJCV VJG KPFWUVTKCN UVTCKP RTQFWEGF TGNCVKXGN[ JKIJ DCUCN NGXGNU QH GZRTGUUKQP QH KVU QYP GPFQIGPQWU OCNVCUG CPF OCNVQUG RGTOGCUG IGPGU. 6JKU RTGUWOCDN[ CNNQYGF VJG [GCUV VQ TCRKFN[ CFCRV VQ OCNVQUG WVKNK\CVKQPÿ UKPEG TGNCVKXGN[ JKIJ NGXGNU QH VJG OCNVQUG RGTOGCUG WPFGT DCUCN EQPFKVKQPU YQWNF CNNQY TCRKF KORQTV QH OCNVQUG VQ KPFWEG HWTVJGT GZRTGUUKQP QH VJG MAL U[UVGO. óJGP VJG FKXGTIGPV MAL RTQOQVGTU HTQO KPFWUVTKCN [GCUV UVTCKPU YGTG GZCOKPGFÿ KV YCU FKUEQXGTGF VJCV UGXGTCN VCPFGON[ TGRGCVGF Á1ñ0 DR TGIKQPU YGTG RTGUGPV KP VJG MAL RTQOQVGTU ($GNN GV CN. 199û). 2TQOQVGT TGIKQPU YGTG ENQPGF VJCV RQUUGUUGF QPG EQR[ÿ VJTGG EQRKGUÿ HQWT EQRKGU CPF HKXG EQRKGU QH VJG VCPFGO TGRGCVU. òWTVJGT KPXGUVKICVKQPU KPFKECVGF VJCV VJG TGRGCVU YGTG PQV RTGUGPV KP VJG NCDQTCVQT[ UVTCKPUÿ DWV YGTG HQWPF KP DCMKPI CPF FKUVKNNKPI UVTCKPU QH [GCUV. #PCN[UKU QH VJG GHHGEV QH VJG VCPFGO TGRGCVU QP IGPG GZRTGUUKQP KPFKECVGF VJCV VJG[ TGFWEGF GZRTGUUKQP QH VJG OCNVQUG RGTOGCUG TGNCVKXG VQ VJG OCNVCUG IGPGÿ KP C HCUJKQP VJCV YCU FGRGPFGPV WRQP VJG JQUV MAL TGIWNCVQT[ DCEMITQWPF CPF VJG PWODGT QH TGRGCVU. +P C JQUV UVTCKP YKVJ VKIJV INWEQUG TGRTGUUKQP CPF UVTQPI OCNVQUG KPFWEVKQPÿ VJG[ OQFGTCVGN[ TGFWEGF (ñ0ð) HWNN[ KPFWEGF GZRTGUUKQP QH VJG OCNVQUG RGTOGCUG IGPGÿ YKVJQWV JCXKPI CP[ CRRCTGPV GHHGEV QP VJG GZRTGUUKQP QH VJG OCNVCUG. *QYGXGTÿ KP C JQUV UVTCKP YKVJ INWEQUG KPUGPUKVKXG MAL IGPG GZRTGUUKQPÿ VJG[ FTCOCVKECNN[ TGFWEGF (WR VQ Áù0-HQNF) HWNN[ KPFWEGF GZRTGUUKQP QH VJG OCNVQUG RGTOGCUG IGPGÿ YKVJQWV JCXKPI CP[ CRRCTGPV GHHGEV QP VJG GZRTGUUKQP QH VJG OCNVCUG ($GNN GV CN. 199û). 2.4.1.6 Improvement of maltose utilization - a complex issue # FKCITCO QH EQPVTQN QH OCNVQUG WVKNK\CVKQP KP [GCUVU KU UJQYP KP òKI. í. 6JG WRVCMG QH J[FTQN[UKU CPF OGVCDQNKUO QH OCNVQUG JCU VQ DG ECTGHWNN[ DCNCPEGF D[ EGNNU KP QTFGT VQ CVVCKP CPF UWUVCKP GHHKEKGPV HGTOGPVCVKQP TCVGU QP VJKU UWICT. òTQO VJG EQORQUKVG FCVC CXCKNCDNG QP OCNVQUG IGPGVKEU KV CRRGCTU VJCV TCRKF TCVGU QH OCNVQUG CFCRVCVKQP CTG CEJKGXGF D[ RQUUGUUKPI OWNVKRNG MAL NQEKÿ YJKEJ KP VWTP CNNQY JKIJ NGXGNU QH GZRTGUUKQP QH VJG OCNVQUG RGTOGCUG CPF OCNVCUG VQ DG QDVCKPGF. óKVJKP VJGUG OWNVKRNG MAL NQEK CTG MAL TGIWNCVQT[ IGPGU VJCV XCT[ KP VJGKT UGPUKVKXKV[ VQ INWEQUG TGRTGUUKQPÿ CPF VJWU GZRTGUUKQP QH VJG MAL IGPGU QEEWTU CV C NQY DWV UKIPKHKECPV NGXGN WPFGT INWEQUG TGRTGUUKPI EQPFKVKQPU. #U C TGUWNVÿ OCNVQUG ECP DG KORQTVGF TCRKFN[ KPVQ VJG EGNN YJGPGXGT KV KU RTGUGPV. 6JG KORQTVGF OCNVQUG FKTGEVN[ KPVGTCEVU YKVJÿ CPF CEVKXCVGUÿ VJG MAL TGIWNCVQT[ RTQVGKPÿ VJWU CNNQYKPI TCRKF CFCRVCVKQP VQ OCNVQUG. úPEG VJG KPFWUVTKCN [GCUV KU HWNN[ CFCRVGF VQ OCNVQUG WVKNK\CVKQPÿ KV TWPU VJG TKUM QH OCNVQUG RQKUQPKPI FWG VQ VJG RTGUGPEG QH VJG EQPUVKVWVKXG /CN4. +P [GCUV YKVJ VKIJVN[ TGIWNCVGF MAL TGIWNCVQT[ RTQVGKPUÿ VJKU OCNVQUG RQKUQPKPI KU PQTOCNN[ CXQKFGF D[ C HGGFDCEM NQQR VJCV QEEWTU YJGP VJG OCNVQUG KU J[FTQN[\GF VQ INWEQUG. +H GZEGUUKXG NGXGNU QH OCNVQUG CTG DGKPI KORQTVGF CPF INW-
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
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EQUG CEEWOWNCVGU YKVJKP VJG EGNNÿ VJG INWEQUG ECP DQVJ FKTGEVN[ KPCEVKXCVG VJG MAL TGIWNCVQT[ RTQVGKPÿ CPF CEVKXCVG VJG /KI1 INWEQUG TGRTGUUKQP U[UVGO VQ FKTGEVN[ TGRTGUU GZRTGUUKQP QH DQVJ VJG OCNVCUG CPF OCNVQUG RGTOGCUG. #U C TGUWNVÿ VJG COQWPV QH OCNVQUG KORQTVGF KPVQ VJG EGNN YKNN DG TGFWEGF CU NGUU OCNVQUG RGTOGCUG KU U[PVJGUK\GF. +P KPFWUVTKCN [GCUVÿ VJG RTGUGPEG QH EQPUVKVWVKXG MAL TGIWNCVQT[ RTQVGKPU YKNN GPUWTG VJCV QPN[ VJG /KI1R-FGRGPFGPV INWEQUG TGRTGUUKQP RCVJYC[ KU HWNN[ HWPEVKQPCN. 6JG VCPFGO TGRGCVU YKVJKP VJG KPFWUVTKCN [GCUV UVTCKP MAL RTQOQVGTU JCXG DGGP UJQYP VQ QRGTCVG OQUV GHHGEVKXGN[ KP UVTCKPU YKVJ EQPUVKVWVKXG MAL GZRTGUUKQPÿ CPF VJKU YKNN GPJCPEG VJG UGNGEVKXG TGFWEVKQP QH GZRTGUUKQP QH VJG OCNVQUG RGTOGCUG KP VJGUG [GCUVU. +V CRRGCTU VJGTGHQTGÿ VJCV VJG VCPFGON[ TGRGCVGF TGIKQPU HQWPF YKVJKP VJG MAL FKXGTIGPV RTQOQVGTU OC[ RTQXKFG VJG KPFWUVTKCN UVTCKPU YKVJ CP CDKNKV[ VQ QXGTEQOG VJG RTQDNGOU QH OCNVQUG VQZKEKV[ FWG GZEGUUKXG KORQTV QH OCNVQUG.
(KI. ý. %QPVTQN QH OCNVQUG WVKNK\CVKQP KP [GCUVU. ©HHKEKGPV CPF UWUVCKPGF OGVCDQNKUO QH OCNVQUG KU FGRGPFGPV WRQP KPVGTCEVKQPU DGVYGGP VJG OCNVQUG TGIWNCVQT (/CN4)ÿ OCNVQUG RGTOGCUG (/CN6) CPF OCNVCUG QT ½-INWEQUKFCUG (/CN5) CPF QVJGT HWPEVKQPUÿ KPENWFKPI /KI1. òGGFDCEM EQPVTQN CHHGEVU CEVKXKVKGU QH VJG MG[ RTQVGKPU. +P JKIJN[ CFCRVGF KPFWUVTKCN UVTCKPU VJG MAL CNNGNGU CPF DTQCFGT IGPGVKE DCEMITQWPFU JCXG DGGP UGNGEVGFÿ XKC ENCUUKECN IGPGVKEUÿ VQ CXQKF OCNVQUG RQKUQPKPIÿ YJKNUV OCKPVCKPKPI JKIJ IN[EQN[VKE HNWZ. LKPGU GPFKPI KP CTTQYU KPFKECVG 'CEVKXCVKQP'þ NKPGU GPFKPI KP DCTU KPFKECVG KPJKDKVKQP QT TGRTGUUKQP. 5GG VGZV HQT FGVCKNU.
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
òTQO VJGUG KPXGUVKICVKQPU KPVQ VJG OQNGEWNCT IGPGVKEU QH OCNVQUG OGVCDQNKUO KP KPFWUVTKCN [GCUV UVTCKPUÿ KV KU ENGCT VJCV CNVJQWIJ QPN[ MAL S, T CPF R CTG PGEGUUCT[ HQT OCNVQUG OGVCDQNKUOÿ C OWEJ OQTG EQORNGZ IGPGVKEU JCU DGGP FGXGNQRGF VQ QDVCKP CPF DCNCPEG VJG TCRKF KPFWEVKQP CPF JKIJ HGTOGPVCVKQP TCVGU CEJKGXGF KP EQOOGTEKCNN[ CRRNKGF [GCUV UVTCKPU. 6JG QRVKOK\GF IGPGVKE DCEMITQWPF VJCV GPCDNGU OCNVQUG HGTOGPVCVKQP VQ RTQEGGF GHHKEKGPVN[ KU PQV HWNN[ WPFGTUVQQF CPF YCU QPN[ CEJKGXGF XKC DTGGFKPI CPF UGNGEVKQP UVTCVGIKGU. úP VJG QVJGT JCPFÿ UVTCVGIKGU VJCV WVKNK\GF Tøý# CRRTQCEJGU VQ KPETGCUG GZRTGUUKQP QH MALS CPF MALT "UKORN[" JCXG PQV [KGNFGF EQOOGTEKCNN[ WUGHWN UVTCKPU. 6JKU GZCORNG JKIJNKIJVU VJG XCNWG QH WUKPI OQNGEWNCT DKQNQI[ VQ WPFGTUVCPF VJG HWPFCOGPVCN DCUKU QH KORQTVCPV VTCKVUÿ YJKNUV GORNQ[KPI ENCUUKECN IGPGVKEU VQ QDVCKP EQOOGTEKCNN[ WUGHWN UVTCKPU.
2.5 %QPENWUKQPU CPF RTQURGEVU +V KU CRRCTGPV HTQO IGPGVKE CPF OQNGEWNCT UVWFKGU QH NCDQTCVQT[ UVTCKPU QH Saccharomyces VJCV VJG NGXGN QH EQORNGZKV[ KP VJGUG QTICPKUOU KU JKIJ. *QYGXGTÿ VJG UKVWCVKQP YKVJ KPFWUVTKCNN[ TGNGXCPV [GCUV UVTCKPU KU UGXGTCN VKOGU OQTG EQORNGZ. ôGPQOGU QH KPFWUVTKCN [GCUV UVTCKPU CTG NGUU RTGFKEVCDNG YKVJ EQORNGZ RNQKFKGU KPENWFKPI CPGWRNQKFKGU DGKPI EQOOQP. ©XGP YKVJKP URGEKGU VJGTG CTG FKXGTUG IGPQOGUÿ OCFG OQTG EQORNGZ KP UQOG ECUGU D[ RTQEGUUGU QH RGTOKUUKXG URQTWNCVKQP CPF UGNH-OCVKPI. ôGPGVKEU QH KPFWUVTKCN [GCUV UVTCKPU KU GXGP OQTG EQORNGZ VJCP UVWFKGU QH NCDQTCVQT[ UVTCKPU YQWNF UWIIGUV. 6JWUÿ KPFWUVTKCN UVTCKPU GZJKDKV JKIJN[ XCTKCDNG URQTWNCVKQP HTGSWGPEKGU VJCV ECP DG UGNH-OCVKPI QT UVGTKNG. òWPEVKQPCN IGPGVKE UVWFKGU KPFKECVG JKIJN[ EQORNKECVGF CPF UQRJKUVKECVGF EQPVTQN RTQEGUUGU QEEWT KP [GCUV EGNNUÿ UWEJ VJCV OCP[ IGPGU QRGTCVG CETQUU OQTG VJCP QPG HWPEVKQPCN ECVGIQT[ÿ CPF OCP[ OWVCVKQPU CRRGCT VQ DG EQORGPUCVGF HQT D[ CEVKXKVKGU QH IGPGU YKVJ RTGXKQWUN[ WPMPQYP HWPEVKQPU. 7PFQWDVGFN[ VJKU UKVWCVKQP YKNN DG OQTG EQORNGZ YJGTG VJG RNQKFKGU QH KPFWUVTKCN [GCUVU CTG EQPEGTPGF. 4GICTFNGUU QH VJG FKHHKEWNVKGU VJCV VJG EQORNGZ IGPGVKE OCMGWR QH KPFWUVTKCN [GCUVU RTGUGPVÿ ENCUUKECN IGPGVKE CRRTQCEJGU QH OCVKPIÿ OWVCVKQPÿ URJGTQRNCUV HWUKQP CPF E[VQFWEVKQP JCXG HQWPF WUG KP UVTCKP KORTQXGOGPVÿ CPF YKNN EQPVKPWG VQ FQ UQ. óJGTGCU Tøý# VGEJPKSWGU ECP JGNR VQ FGHKPG VJG OQNGEWNCT DCUKU QH KORQTVCPV KPFWUVTKCN VTCKVU KP [GCUVUÿ YG CTG HKPFKPI VJCV OCP[ KORQTVCPV RJGPQV[RGU CTG IQXGTPGF D[ VQQ OCP[ IGPGU KP VQQ UWDVNG C HCUJKQP HQT "UKORNG" IGPGVKE GPIKPGGTKPI VQ DG CRRNKGF. #FFGF VQ VJKU KU VJG RTQDNGO QH RQNKVKECN CPF RWDNKE CEEGRVCPEG QH Tøý# UVTCKPU. +P VJKU ECUGÿ ENCUUKECN IGPGVKEU (YKVJ OCVKPI DGKPI RTGHGTCDNG KH RQUUKDNG) QHHGTU VJG CDKNKV[ VQ TGEQODKPG OWVCVKQPU CPF QVJGT IGPQOG EJCPIGU CV JKIJ TCVGUÿ UQ KORTQXKPI VJG RTQDCDKNKV[ QH CEJKGXKPI FGUKTGF QWVEQOGU.
ýGHGTGPEGU #PIGNQX #+ÿ îCTCFjQX ô+ÿ 4QUJMQXC <ô (199õ) 5VTCKPU UGNGEVKQP QH DCMGT'U [GCUV YKVJ KORTQXGF VGEJPQNQIKECN RTQRGTVKGU. òQQF 4GU +PV ù9:ù3ñ-ù39
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$CEMJWU L©ÿ øG4KUK ,ÿ $TQYP 2úÿ $KUUQP Lò (ù001) òWPEVKQPCN IGPQOKE CPCN[UKU QH C EQOOGTEKCN YKPG UVTCKP QH Saccharomyces cerevisiae WPFGT FKHHGTKPI PKVTQIGP EQPFKVKQPU. ò©/5 ;GCUV 4GU 1:111-1ùñ $CK ;ÿ îQJNCYÿ ô$ (1991) /CPKRWNCVKQP QH VJG '\KPE ENWUVGT' TGIKQP QH VTCPUETKRVKQPCN CEVKXCVQT LEU3 D[ UKVG-FKTGEVGF OWVCIGPGUKU. ýWEN #EKF 4GU 19:ñ991-ñ99û $CMCNKPUM[ #6ÿ 5PQY 4 (1990) 6JG EJTQOQUQOCN EQPUVKVWVKQP QH YKPG UVTCKPU QH Saccharomyces cerevisiae. ;GCUV õ:3õû-3÷ù $CPGTjGG ýÿ <JCPI /ê (ù00ù) òWPEVKQPCN IGPQOKEU CU CRRNKGF VQ OCRRKPI VTCPUETKRVKQP TGIWNCVQT[ PGVYQTMU. %WTT úRKP /KETQDKQN ñ:313-31û $CTVJ ôÿ ôCKNNCTFKP % (199û) 2J[UKQNQI[ CPF IGPGVKEU QH VJG FKOQTRJKE HWPIWU Yarrowia lipolytica. ò©/5 /KETQDKQN 4GX 19:ù19-ù3û $CTTG 2ÿ 8o\KPJGV òÿ øGSWKP 5ÿ $NQPFKP $ (1993) ôGPGVKE KORTQXGOGPV QH YKPG [GCUV. +P: òNGGV ô* (GF) óKPG OKETQDKQNQI[ CPF DKQVGEJPQNQI[. *CTYQQF #ECFGOKE 2TGUUÿ 4GCFKPIÿ 7îÿ RR íù1-ííû. $CUUGN ,ÿ óCTHGN ,ÿ /QTVKOGT 4î (19û1) %QORNGOGPVCVKQP CPF IGPGVKE TGEQODKPCVKQP KP Candida lipolytica. , $CEVGTKQN 10÷:õ09-õ11 $CUTCK /#ÿ *KGVGT 2ÿ $QGMG ,ø (199û) 5OCNN QRGP TGCFKPI HTCOGU: DGCWVKHWN PGGFNGU KP VJG JC[UVCEM. ôGPQOG 4GU û:ûõ÷-ûû1 $GNN 2,Lÿ øGGTG øÿ 5JGP ,ÿ %JCROCP $ÿ $KUUKPIGT 2*ÿ #VVHKGNF 28ÿ 8GCN ø# (199÷) # HNQY E[VQOGVTKE OGVJQF HQT TCRKF UGNGEVKQP QH PQXGN KPFWUVTKCN [GCUV J[DTKFU. #RRN ©PXKTQP /KETQDKQN õí:1õõ9-1õûù $GNN 2,Lÿ *KIIKPU 8,ÿ øCYGU +óÿ $KUUKPIGT 2* (199û) 6CPFGON[ TGRGCVGF 1íûD DR GNGOGPVU ECWUG UVTWEVWTCN CPF HWPEVKQPCN XCTKCVKQP KP FKXGTIGPV MAL RTQOQVGTU QH Saccharomyces cerevisiae. ;GCUV 13:113ñ-11íí $GNN 2,Lÿ *KIIKPU 8,ÿ #VVHKGNF 28 (ù001) %QORCTKUQP QH HGTOGPVCVKXG ECRCEKVKGU QH KPFWUVTKCN DCMKPI CPF YKNF-V[RG [GCUVU QH VJG URGEKGU Saccharomyces cerevisiae KP FKHHGTGPV UWICT OGFKC. LGVVU #RRN /KETQDKQN 3ù:ùùí-ùù9 $GNNQEJ %ÿ òGTPhPFG\-©URKPCT 6ÿ êWGTQN #ÿ ôCTEsC /øÿ $CTTKQ © (ù00ù) #P CPCN[UKU QH KPVGT- CPF KPVTCURGEKHKE IGPGVKE XCTKCDKNKVKGU KP VJG Kluyveromyces marxianus ITQWR QH [GCUV URGEKGU HQT VJG TGEQPUKFGTCVKQP QH VJG K lactis VCZQP. ;GCUV 19:ùñû-ùõ÷ $GPsVG\ 6ÿ /CTVsPG\ 2ÿ %QFxP #% (199õ) ôGPGVKE EQPUVKVWVKQP QH KPFWUVTKCN [GCUV. /KETQDKQNQIKC 1ù:3û1-3÷í $KCPEJK //ÿ ýIQ 5ÿ 8CPFGPDQN /ÿ 5CTVQP ôÿ /QTNWRK #ÿ 4KEEK %ÿ 5VCHCPK 5ÿ /QTNKPQ ô$ÿ *KNIGT òÿ %CTKIPCPK ôÿ 5NQPKOUMK 22ÿ òTQPVCNK L (ù001) LCTIG-UECNG RJGPQV[RKE CPCN[UKU TGXGCNU KFGPVKECN EQPVTKDWVKQPU VQ EGNN HWPEVKQPU QH MPQYP CPF WPMPQYP [GCUV IGPGU. ;GCUV 1÷:139û-1í1ù $KFGPPG %ÿ $NQPFKP $ÿ øGSWKP 5ÿ 8o\KPJGV ò (199ù) #PCN[UKU QH VJG EJTQOQUQOCN øý# RQN[OQTRJKUO QH YKPG UVTCKPU QH Saccharomyces cerevisiae. %WTT ôGPGV ùù:1-û $NCPFKP ôÿ ú\KGT-îCNQIGTQRQWNQU úÿ óKPEMGT 2ÿ #TVKIWGPCXG òÿ øWjQP $ (ù000) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: 1õ. Candida tropicalis. ò©$5 LGVV í÷û:91-9í $NQODGTI # (199û) úUOQTGURQPUKXG RTQVGKPU CPF HWPEVKQPCN CUUGUUOGPV UVTCVGIKGU KP Saccharomyces cerevisiae. ©NGEVTQRJQTGUKU 1÷:1íù9-1íí0 $QNQVKP-òWMWJCTC /ÿ 6QHHCPQ-ýKQEJG %ÿ #TVKIWGPCXG òÿ øWEJCVGCW-ýIW[GP ôÿ LGOCKTG /ÿ /CTOGKUUG 4ÿ /QPVTQEJGT 4ÿ 4QDGTV %ÿ 6GTOKGT /ÿ óKPEMGT 2ÿ óoUQNQYUMK-LQWXGN / (ù000) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: 11. Kluyveromyces lactis. ò©$5 LGVV í÷û:õõ-û0
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2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
$QP ©ÿ ýGWXoINKUG %ÿ %CUCTGIQNC 5ÿ #TVKIWGPCXG òÿ óKPEMGT 2ÿ #KING /ÿ øWTTGPU 2 (ù000) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: ñ. Saccharomyces bayanus XCT. uvarum. ò©$5 LGVV í÷û:3û-í1 $TGjPKPI ,ÿ ,GURGTUGP L (ù00ù) 2TQVGKP GZRTGUUKQP FWTKPI NCI RJCUG CPF ITQYVJ KPKVKCVKQP KP Saccharomyces cerevisiae. +PV , òQQF /KETQDKQN ûñ:ùû-3÷ %CFG\ ýÿ 4CURQT 2ÿ FG %QEM #óÿ $QGMJQWV 6ÿ 5OKVJ /6 (ù00ù) /QNGEWNCT KFGPVKHKECVKQP CPF IGPGVKE FKXGTUKV[ YKVJKP URGEKGU QH VJG IGPGTC Hanseniaspora CPF Kloeckera. ò©/5 ;GCUV 4GU 1:ùû9-ù÷9 %CUCToIQNC 5ÿ òG[PGTQN %ÿ øKG\ /ÿ òQWTPKGT 2ÿ ôCKNNCTFKP % (199û) ôGPQOKE QTICPK\CVKQP QH VJG [GCUV ;CTTQY NKRQN[VKEC. %JTQOQUQOC 10õ:3÷0-390 %CUCToIQNC 5ÿ ýGWXoINKUG %ÿ LoRNKPING #ÿ $QP ©ÿ òG[PGTQN %ÿ #TVKIWGPCXG òÿ óKPEMGT 2ÿ ôCKNNCTFKP % (ù000) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: 1û. Yarrowia lipolytica. ò©$5 LGVV í÷û:9ñ-100 %CXCNKGTK øÿ $CTDGTKQ %ÿ %CUCNQPG ©ÿ 2KP\CWVK òÿ 5GDCUVKCPK ©ÿ /QTVKOGT 4ÿ 2QKUKPGNNK / (199÷) ôGPGVKE CPF OQNGEWNCT FKXGTUKV[ KP Saccharomyces cerevisiae PCVWTCN RQRWNCVKQP. òQQF 6GEJPQN $KQVGEJPQN 3õ:íñ-ñ0 %JCTTQP /,ÿ /KEJGNU %# (19÷û) 6JG EQPUVKVWVKXGÿ INWEQUG-TGRTGUUKQP-KPUGPUKVKXG OWVCVKQP QH VJG [GCUV MAL4 NQEWU KU CP CNVGTCVKQP QH VJG MAL43 IGPG. ôGPGVKEU 11õ:ù3-31 %JCTTQP /,ÿ /KEJGNU %# (19÷÷) 6JG PCVWTCNN[ QEEWTTKPI CNNGNGU QH MAL1 KP Saccharomyces URGEKGU GXQNXGF D[ XCTKQWU OWVCIGPKE RTQEGUUGU KPENWFKPI EJTQOQUQOCN TGCTTCPIGOGPV. ôGPGVKEU 1ù0: ÷3-93 %JGTT[ ,/ÿ $CNN %ÿ óGPI 5ÿ ,WXKM ôÿ 5EJOKFV 4ÿ #FNGT %ÿ øWPP $ÿ øYKIJV 5ÿ 4KNGU Lÿ /QTVKOGT 4îÿ $QVUVGKP ø (199û) ôGPGVKE CPF RJ[UKECN OCRU QH Saccharomyces cerevisiae. ýCVWTG 3÷û UWRRN: õû-û3 %JQY 6ÿ ôQNFGPVJCN /,ÿ %QJGP ,øÿ *GFIG /ÿ /CTOWT , (19÷3) +FGPVKHKECVKQP CPF RJ[UKECN EJCTCEVGTK\CVKQP QH [GCUV OCNVCUG UVTWEVWTCN IGPGU. /QN ôGP ôGPGV 191: 3õõ-û1 %JQY 6*%ÿ 5QNNKVVK 2ÿ /CTOWT , (19÷9) 5VTWEVWTG QH VJG OWNVKIGPG HCOKN[ QH MAL NQEK KP Saccharomyces cerevisiae. /QN ôGP ôGPGV ù1û: õ0-õ9 %QFxP #%ÿ $GPsVG\ 6ÿ îQTJQNC / (199û) %JTQOQUQOCN TGQTICPK\CVKQP FWTKPI OGKQUKU QH Saccharomyces cerevisiae DCMGT'U [GCUVU. %WTT ôGPGV 3ù:ùíû-ùñ9 %QFxP #%ÿ ôCUGPV-4COsTG\ ,/ÿ $GPsVG\ 6 (199ñ) òCEVQTU YJKEJ CHHGEV VJG HTGSWGPE[ QH URQTWNCVKQP CPF VGVTCF HQTOCVKQP KP Saccharomyces cerevisiae DCMGT'U [GCUVU. #RRN ©PXKTQP /KETQDKQN õ1:õ30-õ3÷ øCP\K 5©ÿ <JCPI $ÿ CPF /KEJGNU %# (ù000) #NVGTCVKQPU KP VJG Saccharomyces MALCEVKXCVQT ECWUG EQPUVKVWVKXKV[ DWV ECP DG UWRRTGUUGF D[ KPVTCIGPKE OWVCVKQPU. %WTT ôGPGV 3÷:ù33-ùí0 øC[ 4©ÿ 4QIGTU 2,ÿ øCYGU +óÿ *KIIKPU 8, (ù00ù) /QNGEWNCT CPCN[UKU QH OCNVQVTKQUG VTCPURQTV CPF WVKNK\CVKQP D[ Saccharomyces cerevisiae. #RRN ©PXKTQP /KETQDKQN õ÷:ñ3ùõñ33ñ øG $CTTQU LQRGU /ÿ $GNNQP ,4ÿ 5JKTNG[ ý,ÿ ôCPVGT 2ò (ù00ù) ©XKFGPEG HQT OWNVKRNG KPVGTURGEKHKE J[DTKFK\CVKQP KP Saccharomyces UGPUW UVTKEVQ URGEKGU. ò©/5 ;GCUV 4GU 1:3ù3331 øG ýQDGN *ÿ LCYTKG Lÿ $TWN 5ÿ îNKU òÿ øCXKU /ÿ #NNQWUJ *ÿ %QQVG 2 (ù001) 2CTCNNGN CPF EQORCTCVKXG CPCN[UKU QH VJG RTQVGQOG CPF VTCPUETKRVQOG QH UQTDKE CEKF-UVTGUUGF Saccharomyces cerevisiae. ;GCUV 1÷:1í13-1íù÷ øGSWKP 5 (ù001) 6JG RQVGPVKCN QH IGPGVKE GPIKPGGTKPI HQT KORTQXKPI DTGYKPIÿ YKPG-OCMKPI CPF DCMKPI [GCUVU. #RRN /KETQDKQN $KQVGEJPQN ñõ:ñûû-ñ÷÷
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*CJP ;5ÿ îCYCK * (1990) +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH HTGG\G-VQNGTCPV [GCUV HTQO PCVWTG CXCKNCDNG HQT VJG HTQ\GP-FQWIJ OGVJQF. #ITKE $KQN %JGO ñí:÷ù9-÷31 *COOQPF ,4/ (199õ) ;GCUV IGPGVKEU. +P: 2TKGUV òôÿ %CORDGNN + (GFU) $TGYKPI /KETQDKQNQI[. %JCROCP CPF *CNNÿ LQPFQPÿ RR íñ-÷ù *CPUGP ,ÿ îKGNNCPF-$TCPFV /% (199õ) /QFKHKECVKQP QH DKQEJGOKECN RCVJYC[U KP KPFWUVTKCN [GCUVU. , $KQVGEJPQN í9:1-1ù *GTUMQYKV\ +ÿ úUJKOC ; (19÷1) %QPVTQN QH EGNN V[RG KP Saccharomyces cerevisiae: /CVKPI V[RG CPF OCVKPI-V[RG KPVGTEQPXGTUKQP. +P: 5TCVJGTP ,ýÿ ,QPGU ©óÿ $TQCEJ ,4 (GFU) 6JG OQNGEWNCT DKQNQI[ QH VJG [GCUV Saccharomyces: NKHG E[ENG CPF KPJGTKVCPEG. %QNF 5RTKPI *CTDQT LCDQTCVQT[ÿ ýGY ;QTMÿ RR 1÷1-ù09 *KIIKPU 8,ÿ $TCKFYQQF /ÿ $GNN 2ÿ $KUUKPIGT 2ÿ øCYGU +óÿ #VVHKGNF 28 (1999C) ôGPGVKE GXKFGPEG VJCV JKIJ PQPKPFWEGF OCNVCUG CPF OCNVQUG RGTOGCUG CEVKXKVKGUÿ IQXGTPGF D[ MALx3-GPEQFGF VTCPUETKRVKQPCN TGIWNCVQTUÿ FGVGTOKPG GHHKEKGPE[ QH ICU RTQFWEVKQP D[ DCMGT•U [GCUV KP WPUWICTGF FQWIJ. #RRN ©PXKTQP /KETQDKQN õñ:õ÷0-õ÷ñ *KIIKPU 8,ÿ $TCKFYQQF /ÿ $KUUKPIGT 2ÿ øCYGU +óÿ #VVHKGNF 28 (1999D) LGW3í32JG UWDUVKVWVKQP KP VJG /CNZ3 RTQVGKP QH Saccharomyces cerevisiae KPETGCUGU VJG EQPUVKVWVKXKV[ CPF INWEQUG KPUGPUKVKXKV[ QH MAL IGPG GZRTGUUKQP. %WTT ôGPGV 3ñ:í91-í9÷ *KIIKPU 8,ÿ $GNN 2,Lÿ øCYGU +óÿ #VVHKGNF 28 (ù001) ôGPGTCVKQP QH C PQXGN Saccharomyces cerevisiae UVTCKP VJCV GZJKDKVU UVTQPI OCNVQUG WVKNK\CVKQP CPF J[RGTQUOQVKE TGUKUVCPEG WUKPI PQPTGEQODKPCPV VGEJPKSWGU. #RRN ©PXKTQP /KETQDKQN õû:í3íõ-í3í÷ *KPQ #ÿ 6CMCPQ *ÿ 6CPCMC ; (19÷û) ýGY HTGG\G-VQNGTCPV [GCUV HQT HTQ\GP FQWIJ RTGRCTCVKQPU. %GTGCN %JGO õí:ùõ9-ùûñ *QPI 5*ÿ /CTOWT , (19÷õ) 2TKOCT[ UVTWEVWTG QH VJG OCNVCUG IGPG QH VJG MAL6 NQEWU QH Saccharomyces cerevisiae. ôGPG í1: ûñ-÷í *QVVKPIGT *ÿ ô[UNGT %ÿ ýKGFGTDGTIGT 2 (199÷) $CMGT•U [GCUV JCXKPI C NQY VGORGTCVWTG KPCEVKXCVKQP RTQRGTV[. 75 RCVGPV PQ. ñÿ÷ùûÿûùí *W <ÿ ýGJNKP ,úÿ 4QPPG *ÿ /KEJGNU %# (199ñ) MIG1-FGRGPFGPV CPF MIG1-KPFGRGPFGPV INWEQUG TGIWNCVKQP QH MAL IGPG GZRTGUUKQP KP Saccharomyces cerevisiae. %WTT ôGPGV ù÷:ùñ÷-ùõõ *WIJGU 64 YKVJ ù1 QVJGTU (ù000) òWPEVKQPCN FKUEQXGT[ XKC C EQORGPFKWO QH GZRTGUUKQP RTQHKNGU. %GNN 10ù:109-1ùõ +PIG-8GEJVQOQX 5ôÿ 4GRPGXUMCKC /8ÿ îCTRQXC 65 (19÷õ) *[DTKFK\CVKQP QH EGNNU QH VJG UCOG OCVKPI V[RG KP 5CEEJCTQO[EGU [GCUVU. ôGPGVKMC ùù:ùõùñ-ùõ3õ +VQ 6ÿ %JKDC 6ÿ ú\CYC 4ÿ ;QUJKFCÿ *CVVQTK /ÿ 5CMCMK ; (ù001) # EQORTGJGPUKXG VYQJ[DTKF CPCN[UKU VQ GZRNQTG VJG [GCUV RTQVGKP KPVGTCEVQOG. 2TQE ýCVN #ECF 5EK 75# 9÷:íñõ9-íñûí +VQ 6ÿ 6CUJKTQ îÿ /WVC 5ÿ ú\CYC 4ÿ %JKDC 6ÿ ýKUJK\CYC /ÿ ;COCOQVQ îÿ îWJCTC 5ÿ 5CMCMK ; (ù000) 6QYCTF C RTQVGKP-RTQVGKP KPVGTCEVKQP OCR QH VJG DWFFKPI [GCUV: C EQORTGJGPUKXG U[UVGO VQ GZCOKPG VYQ-J[DTKF KPVGTCEVKQPU KP CNN RQUUKDNG EQODKPCVKQPU DGVYGGP VJG [GCUV RTQVGKPU. 2TQE ýCVN #ECF 5EK 75# 9û:11í3-11íû ,CEQDUQP ôîÿ 6TKXGFK ý$ (1990) ;GCUV UVTCKPUÿ OGVJQF QH RTQFWEVKQP CPF WUG KP DCMKPI. 75 RCVGPV í 9û3 ñõ0 ,CXCFGMCT 85ÿ 5KXC4COCP *ÿ ôQMJCNG ø8 (199ñ) +PFWUVTKCN [GCUV UVTCKP KORTQXGOGPV: EQPUVTWEVKQP QH C JKIJN[ HNQEEWNGPV [GCUV YKVJ C MKNNGT EJCTCEVGT D[ RTQVQRNCUV HWUKQP. , +PF /KETQDKQN 1ñ:9í-10ù ,QJPUVQP ,4 (1990) $TGYKPI CPF FKUVKNNKPI [GCUVU. +P: 5RGPEGT ,ò6ÿ 5RGPEGT ø/ (GFU) ;GCUV VGEJPQNQI[. 5RTKPIGT-8GTNCIÿ $GTNKP *GKFGNDGTIÿ RR ññ-10í
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/QTVKOGT 4îÿ ,QJPUVQP ,4 (19÷õ) ôGPGCNQI[ QH RTKPEKRNG UVTCKPU QH VJG ;GCUV ôGPGVKE 5VQEM %GPVGT. ôGPGVKEU 113:3ñ-í3 /QTVKOGT 4îÿ 4QOCPQ 2ÿ 5W\\K ôÿ 2QNUKPGNNK / (199í) ôGPQOG TGPGYCN: # PGY RJGPQOGPQP TGXGCNGF HTQO C IGPGVKE UVWF[ QH í3 UVTCKPU QH Saccharomyces cerevisiae FGTKXGF HTQO PCVWTCN HGTOGPVCVKQP QH ITCRG OWUVU. ;GCUV 10:1ñí3-1ññù /QTVKOGT 4îÿ 5EJKNF ø (19÷1) ôGPGVKE OCRRKPI KP Saccharomyces cerevisiae. +P: 5TCVJGTP ,ýÿ ,QPGU ©óÿ $TQCEJ ,4 (GFU) 6JG OQNGEWNCT DKQNQI[ QH VJG [GCUV Saccharomyces: NKHG E[ENG CPF KPJGTKVCPEG. %QNF 5RTKPI *CTDQT LCDQTCVQT[ÿ ýGY ;QTMÿ RR 11-ùõ ýCMCICYC 5ÿ úWEJK î (199í) %QPUVTWEVKQP HTQO C UKPING RCTGPV QH DCMGT'U [GCUV UVTCKPU YKVJ JKIJ HTGG\G VQNGTCPEG CPF HGTOGPVCVKXG CEVKXKV[ KP DQVJ NGCP CPF UYGGV FQWIJU. #RRN ©PXKTQP /KETQDKQN õ0:3í99-3ñ0ù ýCWOQX ô+ÿ ýCWOQX ©5ÿ 5EJP·TGT (ù001) ôGPGVKE EJCTCEVGTK\CVKQP QH VJG PQPEQPXGPVKQPCN [GCUV Hansenula anomala. 4GU /KETQDKQN 1ñù:ññ1-ñõù ýGGFNGOCPÿ 4$ (1991) %QPVTQN QH OCNVCUG U[PVJGUKU KP [GCUV. /QN /KETQDKQN ñ:ù0û9-ù0÷í ýGWXGINKUG %ÿ òGNFOCPP *ÿ $QP ©ÿ ôCKNNCTFKP %ÿ %CUCToIQNC 5 (ù00ù) ôGPQOKE GXQNWVKQP QH VJG NQPI VGTOKPCN TGRGCV TGVTQVTCPURQUQPU KP JGOKCUEQO[EGVQWU [GCUVU. ôGPQOG 4GU 1ù:930-9í3 ýGX\INKCFQXC ú8ÿ øCX[FGPMQ 5ôÿ 5OKTPQXC 6+ÿ 5QKFNC 64 (199÷) %T[RVKE JGVGTQMCT[QPU KP Saccharomyces cerevisiae UVTCKPU: /QFGN GZRGTKOGPVU. ôGPGVKMC 3í:1õ03-1õ09 ýGX\INKCFQXC ú8ÿ ôCKXQTQPUMKK ##ÿ #TGVGOQX #$ÿ 5QKFNC 62 (ù00ù) òCEVQTU CHHGEVKPI VJG HTGSWGPE[ QH EQPEGCNGF JGVGTQMCT[QUKU KP Saccharomyces cerevisiae. ôGPGVKMC 3÷:ù93ù99 úFC ;ÿ úWEJK î (1990) *[DTKFK\CVKQP QH DCMGT'U [GCUV D[ VJG TCTG-OCVKPI OGVJQF VQ KORTQXG NGCXGPKPI CDKNKV[ KP FQWIJ. ©P\[OG /KETQDKQN 6GEJPQN 1ù:9÷9-993 úNGUGP îÿ òGNFKPI 6ÿ ôjGTOCPUGP %ÿ *CPUGP , (ù00ù) 6JG F[PCOKEU QH VJG Saccharomyces carlsbergensis DTGYKPI [GCUV VTCPUETKRVQOG FWTKPI C RTQFWEVKQP-UECNG NCIGT DGGT HGTOGPVCVKQP. ò©/5 ;GCUV 4GU ù:ñõ3-ñû3 úNKXGT 5ô (199õ) òTQO øý# UGSWGPEKPI VQ DKQNQIKECN HWPEVKQP. ýCVWTG 3û9:ñ9û-õ00 úNKXGT 5ô (ù00ù) òWPEVKQPCN IGPQOKEU: NGUUQPU HTQO [GCUV. 2JKN 6TCPU 4 5QE LQPF $ 3ñû:1û-ù3 úNKXGT 5ôÿ óKPUQP /îÿ îGNN ø$ÿ $CICP\ ò (199÷) 5[UVGOCVKE HWPEVKQPCN CPCN[UKU QH VJG [GCUV IGPQOG. 6TGPF $KQVGEJPQN 1õ:3û3-3û÷ úPKUJK * (1990) ;GCUVU KP HGTOGPVGF HQQFU. +P: 5RGPEGT ,ò6ÿ 5RGPEGT ø/ (GFU) ;GCUV VGEJPQNQI[. 5RTKPIGT-8GTNCIÿ $GTNKP *GKFGNDGTIÿ RR 1õû-19÷ úWEJK îÿ ýKUJK[C 6ÿ #MK[COC * (19÷3) 78-MKNNGF RTQVQRNCUV HWUKQP CU C OGVJQF HQT DTGGFKPI MKNNGT [GCUVU. , òGTOGPV 6GEJPQN õ1:õ31-õ3ñ 2CTMGT î4ÿ XQP $QTUVGN 4% (19÷û) $CUG-UWDUVKVWVKQP CPF HTCOGUJKHV OWVCIGPGUKU D[ UQFKWO EJNQTKFG CPF RQVCUUKWO EJNQTKFG KP Saccharomyces cerevisiae. /WVCV 4GU 1÷9:11-1í 2GFGTUGP /$ (19÷ñ) øý# UGSWGPEG RQN[OQTRJKUOU KP VJG IGPWU Saccharomyces. ++. #PCN[UKU QH VJG IGPGU RDN1ÿ HIS4ÿ LEU2 CPF 6[ VTCPURQUCDNG GNGOGPVU KP %CTNUDGTIÿ 6WDQTI CPF ùù $CXCTKCP DTGYKPI UVTCKPU. %CTNUDGTI 4GU %QOWP ñ0:ùõ3-ùûù 2GFGTUGP /$ (19÷õ) øý# UGSWGPEG RQN[OQTRJKUOU KP VJG IGPWU Saccharomyces. +++. 4GUVTKEVKQP GPFQPWENGCUG HTCIOGPV RCVVGTPU QH EJTQOQUQOCN TGIKQPU KP DTGYKPI CPF QVJGT [GCUV UVTCKPU. %CTNUDGTI 4GU %QOOWP ñ1:1õ3-1÷3 2oTG\-úTVsP ,©ÿ ôCTEsC-/CTVKPG\ ,ÿ #NDGTQNC 6/ (ù00ù) øý# EJKRU HQT [GCUV DKQVGEJPQNQI[: 6JG ECUG QH YKPG [GCUVU. , $KQVGEJPQN 9÷:ùùû-ùí1
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2GTMKPU ©Lÿ ýGGFNGOCP 4$ (19÷÷) MAL64c KU C INQDCN TGIWNCVQT QH ½-INWEQUKFG HGTOGPVCVKQP: KFGPVKHKECVKQP QH C PGY IGPG KPXQNXGF KP OGNG\KVQUG HGTOGPVCVKQP. %WTT ôGPGV 13:3õ9-3ûñ 2QNCKPC ,ÿ #FCO #%ÿ FGN %CUVKNNQ L (1993) 5GNH-FKRNQKFK\CVKQP KP Saccharomyces cerevisiae kar2 JGVGTQMCT[QPU. %WTT ôGPGV ùí:3õ9-3ûù 2TGVQTKWU +5 (ù000) 6CKNQTKPI YKPG [GCUV HQT VJG PGY OKNNGPPKWO: PQXGN CRRTQCEJGU VQ VJG CPEKGPV CTV QH YKPGOCMKPI. ;GCUV 1õ:õûñ-ûù9 2WKI 5ÿ êWGTQN #ÿ $CTTKQ ©ÿ 2oTTG\-úTVsP ,© (ù000) /KVQVKE TGEQODKPCVKQP CPF IGPGVKE EJCPIGU KP Saccharomyces cerevisiae FWTKPI YKPG HGTOGPVCVKQP. #RRN ©PXKTQP /KETQDKQN õ:ù0ñû-ù0õ1 2WVTCOGPV #ÿ $CTCPQYUMC *ÿ ©jEJCTV #ÿ 2TC\OQ ó (19û÷) /CPICPGUG OWVCIGPGUKU KP [GCUV. /GVJQFU %GNN $KQN ù0:ùñ-3í 2WVTCOGPV #ÿ $CTCPQYUMC *ÿ 2TC\OQ ó (19û3) +PFWEVKQP D[ OCPICPGUG QH OKVQEJQPFTKCN CPVKDKQVKE TGUKUVCPEG OWVCVKQP KP [GCUV. /QN ôGP ôGPGV 1ùõ:3ñû-3õõ 4CEJKFK ýÿ $CTTG 2ÿ $NQPFKP $ (1999) /WNVKRNG 6[-OGFKCVGF EJTQOQUQOCN VTCPUNQECVKQPU NGCF VQ MCT[QV[RG EJCPIGU KP C YKPG UVTCKP QH Saccharomyces cerevisiae. /QN ôGP ôGPGV ùõ1:÷í1-÷ñ0 4CPFG\-ôKN òÿ 5CP\ 2ÿ 2TKGVQ ,# (1999) ©PIKPGGTKPI DCMGT'U [GCUV: TQQO HQT KORTQXGOGPV. 6TGPFU $KQVGEJPQN 1û:ù3û-ùíí 4CPM ô*ÿ %CUG[ÿ ô2ÿ ïKCQ óÿ 2TKPING #6 (1991) 2QN[OQTRJKUO YKVJKP VJG PWENGCT CPF ùËO IGPQOGU QH Saccharomyces cerevisiae. %WTT ôGPGV ù0:1÷9-19í 4GGF ôÿ ýCIQFCYKVJCPC 6ó (1991) ;GCUV VGEJPQNQI[ÿ ùPF GFP. 8CP ýQUVTCPF 4GKPJQNFÿ ýGY ;QTM 4QFKEKQ 4 (19÷õ) +PUGTVKQP QH PQP-JQOQNQIQWU øý# UGSWGPEGU KPVQ C TGIWNCVQT[ IGPG ECWUG C EQPUVKVWVKXG OCNVCUG U[PVJGUKU KP [GCUV. %WTT ôGPGV 11:ù3ñ-ùí1 4QOCPÿ * (19÷1) øGXGNQROGPV QH [GCUV CU CP GZRGTKOGPVCN QTICPKUO. +P: 5TCVJGTP ,ýÿ ,QPGU ©óÿ $TQCEJ ,4 (GFU) 6JG OQNGEWNCT DKQNQI[ QH VJG [GCUV Saccharomyces: NKHG E[ENG CPF KPJGTKVCPEG. %QNF 5RTKPI *CTDQT LCDQTCVQT[ÿ ýGY ;QTMÿ RR 1-10 4QOCPQ 2ÿ 5QNK /ôÿ 5W\\K ôÿ ôTC\KC Lÿ
ñí
2CWN 8. #VVHKGNF CPF 2JKNKR ,. L. $GNN
5JCNQP øÿ 5OKVJ 5,ÿ $TQYP 2ú (199õ) # øý# OKETQCTTC[ U[UVGO HQT CPCN[\KPI EQORNGZ øý# UCORNGU WUKPI VYQ-EQNQWT HNWQTGUEGPV RTQDG J[DTKFK\CVKQP. ôGPQOG 4GU õ:õ39õíñ 5KNNU #/ÿ 2CPEJCN %,ÿ 4WUUGNN +ÿ 5VGYCTV ôô (19÷3) ôGPGVKE OCPKRWNCVKQP QH CO[NQN[VKE GP\[OG RTQFWEVKQP D[ [GCUVU. òQWPF $KQVGEJ +PF òGTO 4GU 1:ù09-ùù÷ 5KTGPMQ ú+ÿ ýGGFNGOCP 4$ (199ñ) 2WTKHKECVKQP CPF DKPFKPI RTQRGTVKGU QH VJG /CNõ3R CEVKXCVQT QH Saccharomyces cerevisiae. %WTT ôGPGV ùû:ñ09-ñ1õ 5PQY 4 (19÷3) ôGPGVKE KORTQXGOGPV QH YKPG [GCUV. +P: 5RGPEGT ,ò6ÿ 5RGPEGT ø/ÿ 5OKVJ #4ó (GFU);GCUV IGPGVKEU - HWPFCOGPVCN CPF CRRNKGF CURGEVU. 5RTKPIGT-8GTNCIÿ ýGY ;QTMÿ RR í39-íñ9 5QNNKVVK 2ÿ /CTOWT , (19÷÷) 2TKOCT[ UVTWEVWTG QH VJG TGIWNCVQT[ IGPG HTQO VJG MAL6 NQEWU QH Saccharomyces carlsbergensis. /QN ôGP ôGPGV ù13:ñõ-õù 5QT òÿ òWMWJCTC * (19÷9) #PCN[UKU QH EJTQOQUQOG RCVVGTPU QH VJG IGPWU Kluyveromyces. ;GCUV ñ:1-10 5QWEKGV ,-L YKVJ ù3 QVJGTU (ù000) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: 1. # UGV QH [GCUV URGEKGU HQT OQNGEWNCT GXQNWVKQP UVWFKGU. ò©$5 LGVVU í÷û:3-1ù 5RGPEGT ø/ÿ 4G[PQNFU ýÿ 5RGPEGT ,ò6 (1990) 2TQVQRNCUV HWUKQPU: CRRNKECVKQP VQ KPFWUVTKCN [GCUVU. +P: 5RGPEGT ,ò6ÿ 5RGPEGT ø/ (GFU) ;GCUV VGEJPQNQI[. 5RTKPIGT-8GTNCIÿ $GTNKP *GKFGNDGTIÿ RR 3í÷-3ñí 5RGPEGT ,ò6ÿ 5RGPEGT ø/ (19÷3) ôGPGVKE KORTQXGOGPV QH KPFWUVTKCN [GCUVU. #PP 4GX /KETQDKQN 3û:1ù1-1íù. 5RGPEGT ,ò6ÿ 5RGPEGT ø/ (1990) ;GCUV VGEJPQNQI[. 5RTKPIGT-8GTNCIÿ $GTNKP *GKFGNDGTI 5VCODWM $7ÿ FG #TCWjQ 25 (ù001) îKPGVKEU QH CEVKXG ½-INWEQUKFG VTCPURQTV KP Saccharomyces cerevisiae. ò©/5 ;GCUV 4GU 1:û3-û÷ 5VGYCTV ôôÿ ,QPGU 4ÿ 4WUUGNN + (19÷ñ) 6JG WUG QH FGTGRTGUUGF [GCUV OWVCPVU KP VJG HGTOGPVCVKQP QH DTGYGT[ YQTV. 2TQE ù0VJ %QPI ©WT $TGY %QPXÿ *GNUKPMKÿ RR ùí3-ùñ0 5[EJTQXC *ÿ $TCWP 8ÿ 2QVGKT 5ÿ 5QWEKGV ,-L (ù000) úTICPK\CVKQP QH URGEKHKE IGPQOKE TGIKQPU QH Zygosaccharomyces rouxii CPF Pichia sorbitophila: EQORCTKUQP YKVJ Saccharomyces cerevisiae. ;GCUV 1õ:13ûû-13÷ñ 6COCOGU ,ÿ %NCTM øÿ *GTTGTQ ,ÿ øQRC\Q ,ÿ $NCUEJMG %ÿ òGTPhPFG\ ,/ÿ úNKXGTQU ,%ÿ 8CNGPEKC # (ù00ù) $KQKPHQTOCVKEU OGVJQFU HQT VJG CPCN[UKU QH GZRTGUUKQP CTTC[U: FCVC ENWUVGTKPI CPF KPHQTOCVKQP GZVTCEVKQP. , $KQVGEJPQN 9÷:ùõ9-ù÷3 6CPIJG #ÿ 6GWPKUUGP #ÿ 8CP øKjEM 2ÿ 6JGXGNGKP ,/ (ù000) +FGPVKHKECVKQP QH IGPGU TGURQPUKDNG HQT KORTQXGF ET[QTGUKUVCPEG KP HGTOGPVKPI [GCUV EGNNU. +PV , òQQF /KETQDKQN ññ:ùñ9-ùõù 6GP $GTIG #/#ÿ
ù ôGPGVKEU CPF ENCUUKECN IGPGVKE OCPKRWNCVKQPU QH KPFWUVTKCN [GCUVU
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7PCNFK /ýÿ #TKMCP $ÿ %QTCN ô (ù00ù) +UQNCVKQP QH CNEQJQN VQNGTCPVÿ QUOQVQNGTCPV CPF VJGTOQVQNGTCPV [GCUV UVTCKPU CPF KORTQXGOGPV QH VJGKT CNEQJQN VQNGTCPEG D[ OWVCIGPGUKU. #EVC /KETQDKQN 2QN ñ1:11ñ-1ù0 7TCPQ ýÿ 5CVQ /ÿ 5CJCTC *ÿ îQUJKPQ 5 (1993C) %QPXGTUKQP QH C PQP-HNQEEWNGPV DTGYGT'U [GCUV VQ HNQEEWNGPV QPGU D[ GNGEVTQHWUKQP. +. +FGPVKHKECVKQP CPF EJCTCEVGTK\CVKQP QH VJG HWUCPVU D[ RWNUG HKGNF IGN GNGEVTQRJQTGUKU. , $KQVGEJPQN ù÷:ù3û-ùíû 7TCPQ ýÿ 5CVQ /ÿ 5CJCTC *ÿ îQUJKPQ 5 (1993D) %QPXGTUKQP QH C PQP-HNQEEWNGPV DTGYGT'U [GCUV VQ HNQEEWNGPV QPGU D[ GNGEVTQHWUKQP. ++. 5OCNN-UECNG DTGYKPI D[ HWUCPVU. , $KQVGEJPQN ù÷:ùí9-ùõ1 8CP øKjEM 2ÿ ôQTYC /-òÿ LGOCKTG îÿ 6GWPKUUGP #ÿ 8GTUGNG /ÿ %QNQODQ 5ÿ øWOQTVKGT òÿ /C 2ÿ 6CPIJG #ÿ LQKG\ #ÿ 6JGXGNGKP ,/ (ù000) %JCTCEVGTK\CVKQP QH C PGY UGV QH OWVCPVU FGHKEKGPV KP HGTOGPVCVKQP-KPFWEGF NQUU QH UVTGUU TGUKUVCPEG HQT WUG KP HTQ\GP FQWIJ CRRNKECVKQPU. +PV , òQQF /KETQDKQN ññ:1÷û-19ù 8KPEGPV 5òÿ $GNN 2,Lÿ $KUUKPIGT 2ÿ ýGXCNCKPGP î/* (1999) %QORCTKUQP QH OGNKDKQUG WVKNK\KPI DCMGT•U [GCUV UVTCKPU RTQFWEGF D[ IGPGVKE GPIKPGGTKPI CPF ENCUUKECN DTGGFKPI. LGVVU #RRN /KETQDKQN ù÷:1í÷-1ñù 8QNEMCGTV ôÿ 8QGV /ÿ 4QDDGP , (199û) 5GSWGPEG CPCN[UKU QH C PGCT-UWDVGNQOGTKE 3ñ.í MD øý# UGIOGPV QP VJG TKIJV CTO QH EJTQOQUQOG 8++ HTQO Saccharomyces cerevisiae ECTT[KPI VJG MAL1 NQEWU TGXGCNU 1ñ EQORNGVG QRGP TGCFKPI HTCOGUÿ KPENWFKPI ZUO1ÿ BGL2 CPF BIO2 IGPGU CPF CP #$% VTCPURQTVGT IGPG. ;GCUV 13:ùñ1-ùñ9 8QP $QTUVGN 4% (1990) /WVCIGPGUKU RTKPEKRNGU CPF UVTCVGIKGU CRRNKGF VQ [GCUV. +P: 5RGPEGT ,ò6ÿ 5RGPEGT ø/ (GFU) ;GCUV VGEJPQNQI[. 5RTKPIGT-8GTNCIÿ $GTNKP *GKFGNDGTIÿ RR 3ññ3õñ óCPI ,ÿ ýGGFNGOCP 4$ (199õ) 4GOQXCN QH C /+ô1R DKPFKPI UKVG EQPXGTVU C MAL63 EQPUVKVWVKXG OWVCPV FGTKXGF D[ EJTQOQUQOCN IGPG EQPXGTUKQP VQ INWEQUG KPUGPUKVKXKV[. ôGPGVKEU 1íù:ñ1-õ3 óKPFKUEJ 5 (19õù) ôGPGVKE [GCUV TGUGCTEJ: OGVJQFU CPF UQOG PGY TGUWNVU. óCNNGTUVGKP %QOO ùí:31õ-3ù3 óKP\GNGT ©#ÿ øCXKU 4ó (199û) òWPEVKQPCN CPCN[UKU QH VJG [GCUV IGPQOG. %WTT úRKP ôGPGV øGX û:ûû1-ûûõ óQFKEMC Lÿ øQPIÿ *ÿ /KVVOCPP /ÿ *Q /-*ÿ LQEMJCTV ø, (199û) ôGPQOG-YKFG GZRTGUUKQP OQPKVQTKPI KP Saccharomyces cerevisiae. ýCVWTG $KQVGEJPQN 1ñ:13ñ9-13õû óQNHG î*ÿ 5JKGNFU ø% (199û) /QNGEWNCT GXKFGPEG HQT CP CPEKGPV FWRNKECVKQP QH VJG GPVKTG [GCUV IGPQOG. ýCVWTG 3÷û:û0÷-û13 óQPI 5ÿ $WVNGT ôÿ óQNHG î* (ù00ù) ôGPG QTFGT GXQNWVKQP CPF RCNGQRQN[RNQKF[ KP JGOKCUEQO[EGVG [GCUVU. 2TQE ýCVN #ECF 5EK 75# 99:9ùûù-9ùûû óW Lòÿ *WIJGU 64ÿ øCXKGTYCNC #2ÿ 4QDKPUQP /øÿ 5VQWIJVQP 4ÿ #NVUEJWNGT 5, (ù00ù) LCTIG-UECNG RTGFKEVKQP QH Saccharomyces cerevisiae IGPG HWPEVKQP WUKPI QXGTNCRRKPI VTCPUETKRVKQPCN ENWUVGTU. ýCVWTG ôGPGV 31:ùññ-ùõñ ;QWPI 6ó (19÷3) 6JG RTQRGTVKGU CPF DTGYKPI RGTHQTOCPEG QH DTGYKPI [GCUVU RQUUGUUKPI MKNNGT EJCTCEVGT. , #O 5QE $TGY %JGO í1:1-í
3 $CMGT•U [GCUV: EJCNNGPIGU CPF HWVWTG RTQURGEVU (TCPEKUEC 4CPFG\-)KN, JCKOG #IWKNGTC, #PVQPKQ %QFxP, #PC /. 4KPExP, (TCPEKUEQ ©UVTWEh CPF JQUG #. 2TKGVQ
#DUVTCEV +P VhG RCUV HGY [GCTU, TGEQODKPCPV &0# VGEhPQNQI[ hCU NGF VQ VhG CRRCTKVKQP QH PGY DCMGT•U [GCUV UVTCKPU, YhKEh hCXG QRVKOK\GF QT PQXGN RTQRGTVKGU, CPF KP VhG PGCT HWVWTG, KV KU GZRGEVGF VhCV VhKU VQQN YKNN RTQFWEG C hWIG URGEVTWO QH URGEKCNK\GF [GCUVU QH hKIh CFFGF XCNWG. 6hGKT KPVTQFWEVKQP KP VhG OCPWHCEVWTKPI OCTMGV YKNN RTQFWEG C FTCOCVKE EhCPIG KP HQTOWNCVKQP, KPITGFKGPVU, QT RTQEGUUKPI EQPFKVKQPU EWTTGPVN[ WUGF KP VhG DCMKPI RTCEVKEG CPF YKNN RTQXKFG PGY RTQFWEVU YKVh GPhCPEGF HNCXQWT, VGZVWTGU, QT GZVGPFGF UhGNH NKHG. #U VhG RQVGPVKCN QH TGEQODKPCPV IGPG GZRTGUUKQP CPF OGVCDQNKE GPIKPGGTKPI KU OQTG WPFGTUVQQF, VhKU VGEhPQNQI[ EQWNF DG HWTVhGT CFFTGUUGF VQ CVVGPF VQ RWDNKE CPF EQPUWOGT FGOCPFU QH GPXKTQPOGPVCNN[ UQWPF RTQEGUUGU CPF hGCNVh[ CPF EQPXGPKGPV RTQFWEVU. 6hKU EhCRVGT TGXKGYU VhG OQUV KORQTVCPV CFXCPEGU KP VhG IGPGVKE KORTQXGOGPV QH DCMGT•U [GCUV, RWVU GORhCUKU QP HWPFCOGPVCN CPF CRRNKGF CURGEVU, CPF FKUEWUUGU RGTURGEVKXGU CPF QWVNQQMU KP VhKU HKGNF.
3.1 +PVTQFWEVKQP $TGCF KU FGHKPGF CU C DCMGF RTQFWEV OCFG HTQO CP[ EGTGCN YKVh KORTQXGF HNCXQWT CPF VGZVWTG TGUWNVKPI HTQO PCVWTCN HGTOGPVCVKQPU. 1DXKQWUN[, VhKU DTQCF FGHKPKVKQP RTGUGPVU C ITGCV XCTKGV[ QH DCMGF IQQFU, YhKEh XCT[ KP HQTOWNCVKQP, KPITGFKGPVU, CPF RTQEGUUKPI EQPFKVKQPU. #NVhQWIh UQOG QH VhGO CTG NGCXGPGF D[ URQPVCPGQWU HGTOGPVCVKQP, OQUV hCXG KP EQOOQP VhCV VhG[ hCXG DCMGT•U [GCUV CU C HWPFCOGPVCN GNGOGPV VQ RTQXKFG C TGRTQFWEKDNG RTQEGUU CPF C hKIh SWCNKV[ RTQFWEV CEEQTFKPI VQ VhG FGOCPFU QH EQPUWOGTU. $CMGT•U [GCUV EQPVTKDWVGU UWDUVCPVKCNN[ VQ VhG V[RKECN DTGCF HNCXQWT VhTQWIh VhG HQTOCVKQP QH XQNCVKNG EQORQWPFU CPF CTQOC RTGEWTUQTU FWTKPI VhG CPCGTQDKE HGTOGPVCVKQP. +V CNUQ CHHGEVU VhG ThGQNQIKECN RTQRGTVKGU QH VhG FQWIh CPF FGXGNQRU VhG ETWOD UVTWEVWTG ÿ4GGF CPF 0CIQFCYKVhCPC þ99þý. $WV KVU RTKOCT[ TQNG KU VhG RTQFWEVKQP QH %1ü, XKC VhG CNEQhQNKE HGTOGPVCVKQP QH UWICTU, YhKEh TGUWNV KP FQWIh NGCXGPKPI. %QOOGTEKCN DCMGT•U [GCUVU CTG FQOGUVKECVGF UVTCKPU QH Saccharomyces cerevisiae VhCV hCXG DGGP UGNGEVGF CPF QRVKOK\GF HQT DCMKPI CRRNKECVKQPU. /QUV QH VhGO CTG hQOQVhCNNKE YKVh C hKIh CPF KTTGIWNCT FGITGG QH RNQKF[ CPF NQY URQTWNCVKQP CDKNKV[ ÿ©XCPU þ99û; %QFxP GV CN. þ998ý. +P CFFKVKQP, VhG[ GZhKDKV EhTQOQUQOCNNGPIVh RQN[OQTRhKUOU CPF TGCTTCPIGF EhTQOQUQOGU YKVh OWNVKRNG VTCPUNQECVKQPU Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
58
(. 4CPFG\-)KN GV CN.
ÿ%QFxP GV CN. þ997ý. 6hGUG URGEKCN EhCTCEVGTKUVKEU hCXG DGGP VhG TGUWNV QH PCVWTCN UGNGEVKQP D[ CFCRVCVKXG GXQNWVKQP VQIGVhGT YKVh VhG UWEEGUUKXG CRRNKECVKQP QH KORTQXGOGPV RTQITCOU. úVTCKP KORTQXGOGPV QH DCMGT•U [GCUV hCU VTCFKVKQPCNN[ TGNKGF QP ENCUUKECN IGPGVKE VGEhPKSWGU KPENWFKPI TCPFQO OWVCIGPGUKU, h[DTKFK\CVKQP CPF RTQVQRNCUV HWUKQP ÿUGG %hCRVGT üý, CPF NCVGT UGNGEVKQP HQT DTQCF VTCKVU, UWEh CU TQDWUVPGUU, ITQYKPI TCVG, QT HGTOGPVCVKXG ECRCEKV[. 6hWU, VhG PCVWTCN CFCRVCVKQP CPF UGNGEVKXG RTGUUWTG GZGTVGF D[ [GCUV RTQFWEGTU HQT OCP[ FGECFGU hCU TGUWNVGF KP VTCPUHQTOKPI, CV C NQY EQUV, CP CDWPFCPV HGGFUVQEM CU OQNCUUGU KP EGNN DKQOCUU QH VGEhPQNQIKECN SWCNKV[. 0GXGTVhGNGUU, VhGTG CTG UVKNN KORQTVCPV VTCKVU VhCV CTG HCT HTQO QRVKOCN. 6hG KORTQXGOGPV QH VhG HGTOGPVCVKXG RGTHQTOCPEG KU UVKNN QPG QH VhG OQUV KORQTVCPV DKQVGEhPQNQIKECN EhCNNGPIGU KP DCMGT•U [GCUV. 6hG [GCUV ICUUKPI TCVG KU ETKVKECN KP DCMKPI VGEhPQNQI[ CPF VhKU FGRGPFU DCUKECNN[ QP KPVTKPUKE EhCTCEVGTKUVKEU QH DCMGT•U [GCUV. 6QNGTCPEG VQ XCTKQWU UVTGUUGU NKMG QUOQVKE QT HTGG\KPI KU ENGCTN[ KPCFGSWCVG. ©ZRQUWTG QH DCMGT•U [GCUV VQ hKIh QUOQVKE RTGUUWTG TGUWNVU KP TCRKF EGNN FGh[FTCVKQP VhCV NKOKVU ITQYVh CPF ICU-RTQFWEVKQP ECRCEKV[ ÿ#VVHKGNF þ997; 4CPFG\-)KN GV CN. þ999ý. %QPUGSWGPVN[, RTQQHKPI VKOG KPETGCUGU CPF VhG DTGCF XQNWOG KU TGFWEGF. 6hKU UKVWCVKQP IGVU YQTUG KP HTQ\GP-UYGGV FQWIh, DGECWUG HTGG\KPI CPF VhCYKPI HWTVhGT TGFWEG VhG YCVGT CEVKXKV[. /QTGQXGT, HTGG\KPI CPF HTQ\GP UVQTCIG QH FQWIh hCU C PGICVKXG KPHNWGPEG QP VhG DCMKPI RGTHQTOCPEG FWG VQ EGNN KPLWT[ D[ HTGG\KPI ÿ#VVHKGNF þ997; 4CPFG\-)KN GV CN. þ999ý. 6Q QXGTEQOG VhGUG RTQDNGOU, KV KU EQOOQP HQT DCMGTU VQ KPETGCUG [GCUV NGXGN KP VhGUG RTQFWEVU D[ ü-8ù CDQXG PQTOCN. *QYGXGT, VhKU RTCEVKEG KPETGCUGU RTQEGUU EQUV CPF PGICVKXGN[ CHHGEVU VhG VCUVG CPF VGZVWTG. %QPUGSWGPVN[, VhG FGXGNQROGPV QH [GCUV UVTCKPU YKVh DGVVGT ICUUKPI RQYGT KP hKIh-UWICT CPF HTQ\GP FQWIh hCU C ITGCV GEQPQOKE KPVGTGUV. %WTTGPVN[, PQ UWEh CRRTQRTKCVG KPFWUVTKCN DCMGT•U [GCUV KU CXCKNCDNG VhCV hCU CNN VhGUG FGUKTGF RTQRGTVKGU, CPF KV KU WPNKMGN[ VhCV ENCUUKECN DTGGFKPI RTQITCOU EQWNF RTQXKFG UKIPKHKECPV KORTQXGOGPVU QH VhGUG EhCTCEVGTKUVKEU. +PFGGF, QWT NKOKVGF WPFGTUVCPFKPI QH Rh[UKQNQIKECN CPF IGPGVKE FGVGTOKPCPVU QH VhGUG VTCKVU UWIIGUVU VhCV VhG[ CTG WPFGT EQPVTQN QH OWNVKRNG IGPGU CPF EQORNGZ TGIWNCVQT[ OGEhCPKUOU ÿ©UVTWEh üûûû; *QhOCPP üûûüý. úQOG QH VhGUG FGUKTGF RhGPQV[RGU EQWNF CNUQ IQ CICKPUV ÉDKQNQIKECN FGUKIPÌ ÿ#VVHKGNF þ997ý. 6hG [GCUV-DCUGF DCMKPI CPF OCPWHCEVWTKPI OCTMGVU CNUQ CHHQTF DKQVGEhPQNQIKECN EhCNNGPIGU FGTKXGF QH VhG ITQYKPI FGOCPFU QH RTQFWEGTU, DCMGTU, CPF EQPUWOGTU HQT PGY GEQPQOKECNN[ CPF GPXKTQPOGPVCNN[ UQWPF RTQEGUUGU CPF EQPXGPKGPV RTQFWEVU ÿ&GSWKP üûûþý. %CPG CPF DGGV OQNCUUGU KPETGCUG KP EQUV [GCT D[ [GCT, HQTEKPI VhG [GCUV RTQFWEGTU VQ GZRNQTG PQXGN CRRTQCEhGU HQT DCMGT•U [GCUV RTQRCICVKQP. 0WVTCEGWVKECNU CTG VhG HCUVGUV-ITQYKPI UGIOGPV QH VQFC[•U HQQF KPFWUVT[. $TGCF, CU C FKCT[ RTQFWEV, KU QPG QH VhG DGUV ECPFKFCVGU VQ DGEQOG C HWPEVKQPCN HQQF. 6hCV KU Yh[ C DCMGT•U [GCUV VhCV RTQFWEGU XKVCOKP RTGEWTUQTU, GUUGPVKCN HCVV[ CEKFU QT RTGDKQVKEU, YQWNF DG YGNEQOG. +P VhKU UEGPCTKQ, VhG CDKNKV[ VQ VTCPUHQTO DCMGT•U [GCUV D[ TGEQODKPCPV &0# VGEhPQNQI[ hCU QRGPGF VhG RQUUKDKNKVKGU VQ OCPKRWNCVG LWUV C UKPING IGPG QT RCVhYC[ YKVhQWV CNVGTKPI QVhGT MG[ QT XCNWCDNG IGPGU HQT VGEhPQNQIKECN RWTRQUGU ÿ4CPFG\-)KN GV CN. þ999; &GSWKP üûûþý. +V KU CNUQ RQUUKDNG, VQ GNKOKPCVG WPFGUKTCDNG IGPGU, VQ GPIKPGGT VhG EQPUVKVWVKXG GZRTGUUKQP QH ITQWRU QH IGPGU KPXQNXGF KP C RCT-
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
59
VKEWNCT HWPEVKQP, QT GXGP, VQ UYKVEh QP QT QHH VhG VTCPUETKRVKQP QH C IKXGP IGPG KP C URGEKHKE VGEhPQNQIKECN UVGR. +P CFFKVKQP, VhG CRRNKECVKQP QH Rh[UKQNQIKECN CPF OQNGEWNCT UVWFKGU KP VhG GPXKTQPOGPV GPEQWPVGTGF D[ DCMGT•U [GCUV ÿ*KIIKPU GV CN. þ999; XCP *QGM GV CN. üûûû; *KIIKPU GV CN. üûûþý CPF VhG KPVTQFWEVKQP QH HWPEVKQPCN IGPQOKE CPF RTQVGQOKE CRRTQCEhGU ÿ6CPIhG GV CN. üûûû; 4QFTKIWG\-øCTICU GV CN. üûûü; UGG CNUQ %hCRVGT þþý KU NGCFKPI VQ VhG KFGPVKHKECVKQP QH VCTIGV IGPGU CPF TGIWNCVQT[ RCVhYC[U KORNKGF KP EQOOGTEKCNN[ KORQTVCPV VTCKVU. 6hG VTCPUHGT QH hGVGTQNQIQWU IGPGU VQ S. cerevisiae hCU CNUQ OGCPV VhCV VhGTG KU PQY VhG RQUUKDKNKV[ QH WUKPI DCMGT•U [GCUV CU C EGNN HCEVQT[. 6hKU hCU CNNQYGF KP VhG RCUV FGECFG VhG FGXGNQROGPV QH KPFWUVTKCN UVTCKPU YKVh WPUWURGEVGF RTQRGTVKGU, IKXKPI TKUG VQ C PGY IGPGTCVKQP QH DCMGT•U [GCUV ÿ4CPFG\-)KN GV CN. þ999; &GSWKP üûûþý. 6hKU EhCRVGT YKNN EQNNGEV VhG CXCKNCDNG KPHQTOCVKQP QP VhG OQUV KORQTVCPV CFXCPEGU KP VhG IGPGVKE KORTQXGOGPV QH DCMGT•U [GCUV, HTQO DQVh HWPFCOGPVCN CPF VGEhPQNQIKECN CURGEVU. ÷G YKNN HQEWU QWT CVVGPVKQP QP KUUWGU TGNCVGF YKVh FQYPUVTGCO CRRNKECVKQPU QH DCMGT•U [GCUV, KP RCTVKEWNCT VQ HGTOGPVCVKXG RGTHQTOCPEG, QUOQVQNGTCPEG, CPF ET[QTGUKUVCPEG. +PHQTOCVKQP EQPEGTPKPI YKNF S. cerevisiae CPF QVhGT KPFWUVTKCN [GCUVU, UWEh CU Torulaspora, YKNN DG CNUQ EQPUKFGTGF YhGTG KV RTQXKFGU MPQYNGFIG VhCV CFFTGUUGU WPTGUQNXGF RTQDNGOU QT TGXGCNU HWVWTG RGTURGEVKXGU CPF CNVGTPCVKXGU. 1VhGT KORQTVCPV VCTIGVU HQT UVTCKP KORTQXGOGPV, NKMG GZVGPUKQP QH UWDUVTCVG TCPIG, hGVGTQNQIQWU GP\[OG GZRTGUUKQP CPF QXGTRTQFWEVKQP QH GUUGPVKCN PWVTKGPVU hCXG TGEGPVN[ DGGP TGXKGYGF ÿ&GSWKP üûûþ; 2TKGVQ GV CN. üûû3ý CPF VhGTGHQTG YKNN DG PQV VTGCVGF hGTG.
3.2 )GPGVKE EJCTCEVGTKUVKEU QH DCMGT•U [GCUV UVTCKPU 6TCFKVKQPCN CRRNKECVKQPU QH KPFWUVTKCN [GCUV KPENWFG VhG HQTOCVKQP QH KORQTVCPV RTQFWEVU, VhG TGUWNV QH URGEKHKE HGTOGPVCVKQP ECTTKGF QWV D[ UVTCKPU QH S. cerevisiae. 6hGUG UVTCKPU UhCTG EQOOQP HGCVWTGU UWEh CU GHHKEKGPV UWICT WVKNK\CVKQP, hKIh GVhCPQN VQNGTCPEG CPF RTQFWEVKQP, CTQOC CPF HNCXQWT HQTOCVKQP, hKIh [KGNF CPF HGTOGPVCVKQP TCVG, CPF IGPGVKE UVCDKNKV[, YhKEh IWCTCPVGG VhG TGRTQFWEKDKNKV[ QH RTQRGTVKGU KP VhG HKPCN RTQFWEVU ÿ$GPsVG\ GV CN. þ99ö; )CUGPV-4COsTG\ GV CN. þ999ý. õKMG OQUV KPFWUVTKCN UVTCKPU QH S. cerevisiae, DCMGT•U [GCUVU CTG RQQTN[ EhCTCEVGTK\GF IGPGVKE GPVKVKGU CPF hCXG NQPI DGGP EQPUKFGTGF CU OKETQQTICPKUOU VhCV RQUUGUU CP WPMPQYP EhTQOQUQOCN EQPUVKVWVKQP CPF C ITGCV FGCN QH IGPGVKE hGVGTQIGPGKV[. +P CFFKVKQP, GCEh KPFWUVTKCN ITQWR RQUUGUUGU QVhGT URGEKHKE RTQRGTVKGU ÿUGG CNUQ %hCRVGT üý. 3.2.1 0WENGCT IGPQOG 3.2.1.1 DNA content úQOG CWVhQTU ÿ#FCOU GV CN. þ99ü; õQWKU GV CN. þ99ô; 2GFGTUGP þ998ý hCXG TGRQTVGF NCTIG XCTKCVKQPU KP VhG &0# EQPVGPV CPF VhG EhTQOQUQOG PWODGT QH FKHHGTGPV KP-
öû
(. 4CPFG\-)KN GV CN.
6CDNG 1. 2NQKF[ (n) QH FKHHGTGPV S. cerevisiae KPFWUVTKCN UVTCKPU úVTCKP
2NQKF[ ÿPý
ó00ü95 ) þ &ú8þ ) ü &#&+ $ þ.5 øú $ ü.5 %6 $ ü.7 ú$ü $ ü.ö ú$þþ $ þ.3 øþ $ ü.7 øü $ ü.7 #%#üþ ÷ þ.9 #%#üü ÷ þ.9 #õò1þüô5 & þ.ö #õò1þ5ü3 & þ.3 #õò17ô3 $ 3 #õò155ô $ þ.3 #õò1þöþþ $ 3 6úþôö $ þ.ö #6%%9û8û $T þ.ö 0%ó%39ö $T ü.ü ) IGPGVKE NKPG; $ DCMGTU•; ÷ YKPG, $T DTGYGTU•; & FKUVKNNGTU•. /QFKHKGF HTQO %QFxP GV CN. ÿþ998ý. 4GRTQFWEGF YKVh RGTOKUUKQP.
FWUVTKCN [GCUV, CPF hCXG UWIIGUVGF VhCV VhGUG XCTKCVKQPU OC[ TGURQPF VQ URGEKHKE KPFWUVTKCN GPXKTQPOGPVU ÿUGG %hCRVGT üý. *QYGXGT, YhGP EQORCTKPI UKOWNVCPGQWUN[ FKHHGTGPV KPFWUVTKCN [GCUV ITQWRU, K.G., DCMGTU•, FKUVKNNGTU•, DTGYGTU• CPF YKPG UVTCKPU, UKOKNCT XCTKCVKQPU KP &0# EQPVGPV hCXG DGGP HQWPF COQPI [GCUV HTQO DQVh VhG UCOG ITQWR CPF HTQO FKHHGTGPV ITQWRU ÿK.G. DCMGTU• XU. DTGYGTU•ý. # ENGCT GZCORNG KU UhQYP KP 6CDNG þ. ÷hGP EQORCTGF YKVh ó00ü95 CPF &ú8þ NCDQTCVQT[ [GCUVU, VhG &0# EQPVGPV HTQO VhG DCMGT•U [GCUVU XCTKGU HTQO þ.3P VQ 3P ÿ%QFxP GV CN. þ998ý. 6hKU KPFKECVGU VhCV, CV NGCUV, &0# EQPVGPV FQGU PQV TGHNGEV URGEKHKE CFCRVCVKQP VQ KPFWUVTKCN GPXKTQPOGPVU. 3.2.1.2 Chromosomal reorganizations %hTQOQUQOCN TGQTICPK\CVKQP KU C PCVWTCN RhGPQOGPQP KP VhG NKHG E[ENG QH OCP[ QTICPKUOU, CPF KP UQOG ECUGU, KV KU RCTV QH VhGKT FGXGNQROGPV RCVVGTP ÿ#IWKNGTC CPF òNGKP þ993ý, HQT KPUVCPEG VhG de novo VGNQOGTG HQTOCVKQP KP S. cerevisiae. +P CFFKVKQP, VhGTG CTG QVhGT PQP-RTQITCOOGF IGPQOG TGQTICPK\CVKQPU, YhKEh KPENWFG FGNGVKQPU, VTCPUNQECVKQPU, GVE. YKVh COC\KPI EQPUGSWGPEGU UWEh CU FTCOCVKE EhCPIGU KP VhG XKCDKNKV[ QH VhG OGKQVKE RTQFWEVU QH C UVTCKP hGVGTQ\[IQWU HQT QPG QH VhGUG TGQTICPK\CVKQPU. +P S. cerevisiae, IGPQOKE TGQTICPK\CVKQP VCMGU RNCEG via TGEQODKPCVKQP, CPF VhKU RhGPQOGPQP QEEWTU DGVYGGP hQOQNQIQWU TGRGCV UGSWGPEGU, YhKEh KP [GCUV, CTG XGT[ UECTEG: VGNQOGTKE CPF UWDVGNQOGTKE ó• CPF ñ TGIKQPU, 6[ VTCPURQUCDNG GNGOGPVU, CPF VQ C NGUUGT GZVGPV, T40# CPF V40# IGPGU ÿ#IWKNGTC CPF
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
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òNGKP þ993ý. $QVh 6[ VTCPURQUCDNG GNGOGPVU CPF ó• UWDVGNQOGTKE TGIKQPU hCXG DGGP FGUETKDGF VQ EQPVTKDWVG VQ EhTQOQUQOG TGQTICPK\CVKQP, KPVTQFWEKPI XCTKCDKNKV[ KP EhTQOQUQOG UK\G CPF PWODGT ÿõQWKU GV CN. þ99ôý. 4GQTICPK\CVKQP CV OGKQUKU hCU DGGP QDUGTXGF DQVh KP CNN VhG VGVTCFU QH VhG UCOG DCMGT•U UVTCKPU CPF KP CNN VhG DCMGT•U [GCUV CPCN[\GF ÿ%QFxP GV CN. þ997ý. 6hG HCEV VhCV VhGUG TGQTICPK\CVKQPU UGGO VQ DG NKOKVGF VQ VhG OGKQVKE RGTKQF KU RCTVKEWNCTN[ YQTVh PQVKEKPI. 6hG PWODGT QH RQN[OQTRhKUOU HQWPF KP VhG GNGEVTQRhQTGVKE MCT[QV[RG RCVVGTPU QH VhG EhTQOQUQOGU QH FKHHGTGPV KPFWUVTKCN [GCUV ITQWRU KU XGT[ hKIh ÿ0CWOQX GV CN. þ99ü; %QFxP GV CN. þ997ý. ÷hGTGCU C UVCPFCTF hCRNQKF NCDQTCVQT[ UVTCKP QH S. cerevisiae ÿQT KVU KUQIGPKE FKRNQKFý FKURNC[U þ5 DCPFU EQTTGURQPFKPI VQ þö EhTQOQUQOGU CPF ITGCV XCTKCVKQPU hCXG DGGP HQWPF KP DCMGT•U [GCUV, YhKEh UhQYGF XCTKCVKQPU KP VhG PWODGT, KPVGPUKV[, CPF RQUKVKQP QH VhG EhTQOQUQOCN DCPFU ÿ%QFxP CPF $GPsVG\ þ995; %QFxP GV CN. þ998ý. 6hGUG FKHHGTGPEGU KP VhG PWODGT QH EhTQOQUQOCN DCPFU CTG OQUVN[ VhG TGUWNV QH hQOQNQIQWU EhTQOQUQOGU QH FKHHGTGPV UK\G. #RCTV HTQO EhCPIGU KP EhTQOQUQOG UK\G, VhGTG CTG CNUQ EhCPIGU KP EhTQOQUQOG PWODGT CU C EQPUGSWGPEG QH NCEM QH RCKTKPI CPF TGEQODKPCVKQP QH hQOQNQIQWU EhTQOQUQOGU ÿYKVh NQY hQOQNQI[ý, CPF PQP-FKULWPEVKQP CV OGKQUKU. 6hG NCVVGT TGUWNVU KP CP WPGSWCN FKUVTKDWVKQP QH VhG EhTQOQUQOGU KP VhG RTQIGP[, NQY XKCDKNKV[, CPF CPGWRNQKFKGU KP VhG UWTXKXKPI RTQFWEVU. #PGWRNQKF[ CPFðQT RQN[RNQKF[ COQPI KPFWUVTKCN [GCUVU KU UQ YKFGURTGCF VhCV VhG OCKPVGPCPEG QH CP WPDCNCPEGF EhTQOQUQOG UGV hCU DGGP UWIIGUVGF VQ DG CFXCPVCIGQWU ÿ#FCOU GV CN. þ99üý. 6hG UGNGEVKQP D[ RTQFWEGTU QH DTQCF [GCUV VTCKVU, UWEh CU TQDWUVPGUU, ITQYKPI TCVG, QT HGTOGPVCVKXG ECRCEKV[ YQWNF hCXG EQPVTKDWVGF VQ VhG IGPGVKE EQPUVKVWVKQP QH DCMGT•U UVTCKPU. #NVhQWIh VhGTG KU C NCEM QH MPQYNGFIG QH VhG IGPGU TGURQPUKDNG HQT VhGUG RTQRGTVKGU, VhG[ hCXG KP EQOOQP VhCV VhG[ CTG HCXQWTGF D[ C hKIh RNQKF[ NGXGN ÿXCP *QGM GV CN. üûûû; *KIIKPU GV CN. üûûþ; CPF %hCRVGT üý. 3.2.1.3 Homology with the DNA of laboratory strains úQOG CWVhQTU hCXG FGVGEVGF KP KPFWUVTKCN [GCUVU, VhG RTGUGPEG QH EhTQOQUQOGU YKVh hKIh QT NQY hQOQNQI[ YhGP EQORCTGF VQ VhQUG QH hCRNQKF TGHGTGPEG NCDQTCVQT[ UVTCKPU. úGSWGPEG FKXGTIGPEG QH hQOQNQIQWU &0# YCU UWHHKEKGPVN[ hKIh VQ RTGXGPV IGPGVKE TGEQODKPCVKQP ÿ2GFGTUGP þ998ý. %hTQOQUQOGU QH KPFWUVTKCN [GCUVU hCXG DGGP h[DTKFK\GF KP CNN ECUGU YKVh RTQDGU HTQO NCDQTCVQT[ UVTCKP IGPGU, VhWU KPFKECVKPI VhCV VhG &0# hQOQNQI[ YKVh VhGUG IGPGU KU XGT[ hKIh ÿ0CWOQX GV CN. þ99ü; %QFxP GV CN. þ995ý. *GPEG, FKHHGTGPV EhTQOQUQOGU YGTG URGEKHKECNN[ FGVGTOKPGF YKVh RTQDGU HQT LEU2, TRP1, URA3, CUP1, SUC2, ADC1 , CPF GAL4 ÿ0CWOQX GV CN. þ99ü; %QFxP GV CN. þ998ý. %hTQOQUQOCN NQECVKQP CRRGCTU VhG UCOG KP DQVh NCDQTCVQT[ CPF KPFWUVTKCN [GCUVU CPF VhG GZKUVGPEG QH hQOQNQIQWU EhTQOQUQOGU QH FKHHGTGPV UK\G YCU EQPHKTOGF ÿ%QFxP GV CN. þ998ý. ©ZEGRVKQPCNN[, VTCPUNQECVKQP VQ QVhGT EhTQOQUQOGU hCU DGGP HQWPF ÿK.G. CUP1 IGPGý ÿ0CWOQX GV CN. þ99ü; %QFxP CPF $GPsVG\ þ995; %QFxP GV CN. þ998ý. (QT SUC2, GPEQFKPI KPXGTVCUG, CPF HQT VhG MAL NQEK, GZVGPUKXG IGPG CORNKHKECVKQP QXGT UGXGTCN EhTQOQUQOGU hCU DGGP UhQYP HQT DCMGT•U CPF FKUVKNNGT•U [GCUV ÿõQWKU GV CN. þ99ô; %QFxP GV CN. þ998; CPF %hCRVGT üý.
öü
(. 4CPFG\-)KN GV CN.
3.2.2 5RQTWNCVKQP +PFWUVTKCN [GCUV UVTCKPU WUWCNN[ URQTWNCVG RQQTN[ CPF TCTGN[ HQTO HQWT-URQTG CUEK. /QTGQXGT, URQTG XKCDKNKVKGU CTG XGT[ NQY ÿUGG CNUQ %hCRVGT üý. 6hKU hCU QHVGP KORGFGF ETQUUDTGGFKPI CPF IGPGVKE EhCTCEVGTK\CVKQP. 6hWU, KORTQXGOGPV QH URQTWNCVKQP CPF HQWT-URQTG CUEWU HQTOCVKQP HTGSWGPEKGU KU C RTGTGSWKUKVG HQT DQVh C FGVCKNGF IGPGVKE CPCN[UKU CPF IGPGVKE KORTQXGOGPV D[ h[DTKFK\CVKQP KP DCMGT•U [GCUV. /CP[ VTGCVOGPVU hCXG DGGP FGUETKDGF VQ CHHGEV VhG HTGSWGPE[ QH URQTWNCVKQP CPF VhG PWODGT QH URQTGU RGT CUEWU. /QUV RTQEGFWTGU VQ KPFWEG URQTWNCVKQP KPXQNXG C RGTKQF QH XGIGVCVKXG ITQYVh ÿRTGURQTWNCVKQP OGFKWOý CPF VhGP VTCPUHGT VQ URQTWNCVKQP OGFKWO. 4GURKTCVQT[ UWHHKEKGPE[ CPF FGTGRTGUUKQP hCXG DGGP FGUETKDGF CU DGKPI RTGTGSWKUKVGU HQT NCDQTCVQT[ UVTCKPU URQTWNCVKQP. +P EQPVTCUV, HTGSWGPEKGU QH URQTWNCVKQP CPF HQWT-URQTG CUEWU HQTOCVKQP WR VQ 97 CPF öûù, TGURGEVKXGN[, YGTG QDVCKPGF KP DCMGT•U UVTCKPU CHVGT VTCPUHGT QH þûù-INWEQUG ITQYP EGNNU VQ URQTWNCVKQP OGFKWO CV üü¸% ÿ%QFxP GV CN. þ995ý. 6hKU UWIIGUVU VhCV KP DCMGT•U UVTCKPU EQORNGVKQP QH TGURKTCVQT[ CFCRVCVKQP QEEWTU FWTKPI VhG HKTUV HGY hQWTU KP URQTWNCVKQP OGFKWO, UQ VhCV VhKU KU UWHHKEKGPV VQ RGTOKV KPFWEVKQP QH hKIh HTGSWGPEKGU QH URQTWNCVKQP. úWEh HTGSWGPEKGU QH URQTWNCVKQP CPF VGVTCF HQTOCVKQP UhQWNF CNNQY HWTVhGT IGPGVKE CPCN[UKU CU YGNN CU IGPGVKE KORTQXGOGPV D[ h[DTKFK\CVKQP.
3.3 +ORQTVCPV VTCKVU HQT DCMKPI CRRNKECVKQPU 3.3.1 (GTOGPVCVKXG EJCTCEVGTKUVKEU $CMGTU• [GCUV KU RTQFWEGF HTQO OQNCUUGU, YhGTG UWETQUG KU VhG OCLQT UWICT. +P ECPG OQNCUUGU, VhGTG KU CNUQ C TGNCVKXGN[ hKIh RTQRQTVKQP QH INWEQUG CPF HTWEVQUG, YhGTGCU VhG COQWPV QH VhGUG TGFWEKPI UWICTU KU NQY KP DGGV OQNCUUGU. úWETQUG KU ENGCXGF QWVUKFG VhG EGNN D[ KPXGTVCUG KPVQ INWEQUG CPF HTWEVQUG, YhKEh CTG VhGP VTCPURQTVGF CETQUU VhG RNCUOC OGODTCPG D[ UGXGTCN hGZQUG VTCPURQTVGTU. $TGCF FQWIh KU TKEh KP OCNVQUG. 6hKU FKUCEEhCTKFG KU VTCPURQTVGF VhTQWIh C OCNVQUG RGTOGCUG CPF KU UWDUGSWGPVN[ h[FTQN[UGF KPVQ INWEQUG D[ OCNVCUG ÿVhG OCNVQUG OGVCDQNKUO KU FGUETKDGF KP %hCRVGT üý. )NWEQUG KU PQV QPN[ VhG RTGHGTTGF ECTDQP UQWTEG HQT [GCUV, KV EQPVTQNU C XCTKGV[ QH OGEhCPKUOU VhCV CEV CV FKHHGTGPV NGXGNU, GPUWTKPI KVU RTKOCT[ WVKNK\CVKQP CPF VhG TGIWNCVKQP QH OGVCDQNKUO, EGNN ITQYVh, CPF FGXGNQROGPV. #OQPI VhG RCVhYC[U TGIWNCVGF D[ INWEQUG, VhG VTCPUETKRVKQPCN TGRTGUUKQP QH IGPGU GPEQFKPI GP\[OGU HQT VhG OGVCDQNKUO QH CNVGTPCVG UWICTU ÿINWEQUG QT ECVCDQNKVG TGRTGUUKQPý CPF VhG 4CU-E#/2 RCVhYC[ CTG VhG DGUV WPFGTUVQQF. òPQYNGFIG QH UWICT OGVCDQNKUO, DQVh CV IGPGVKE CPF DKQEhGOKECN NGXGN, hCU DGGP WUGHWN VQ FGUKIP FKHHGTGPV UVTCVGIKGU KP QTFGT VQ KORTQXG VhG HGTOGPVCVKXG RGTHQTOCPEG QH DCMGTU• [GCUV UVTCKPU.
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
ö3
3.3.1.1 Sugar metabolism, transport and control $CMGT•U [GCUV RTGHGTU VQ WUG INWEQUG VQ CP[ UWICT, CPF KVU RTGUGPEG KPFWEGU C XCTKGV[ QH OGEhCPKUOU VQ RTGXGPV VhG WVKNK\CVKQP QH CNVGTPCVKXG ECTDQh[FTCVGU. 6hGUG OGEhCPKUOU KPENWFG VhG VTCPUETKRVKQPCN TGRTGUUKQP QH C NCTIG PWODGT QH IGPGU, C RhGPQOGPQP MPQYP CU ÉINWEQUG ÿQT ECVCDQNKVGý TGRTGUUKQPÌ ÿ)CPEGFQ þ998; *CTFKG GV CN. þ998; %CTNUQP þ999; JQhPUVQP þ999; /QTGPQ CPF *GTTGTQ üûûüý. 6hG IGPGU TGIWNCVGF D[ INWEQUG TGRTGUUKQP ECP DG ENCUUKHKGF KP VhTGG OCKP ITQWRU ÿ4QPPG þ995ý: IGPGU GPEQFKPI URGEKHKE INWEQPGQIGPKE GP\[OGU, IGPGU GPEQFKPI VhG OKVQEhQPFTKCN GP\[OGU QH VhG òTGDU E[ENG, CPF IGPGU GPEQFKPI RTQVGKPU KPXQNXGF KP VhG WRVCMG CPF OGVCDQNKUO QH CNVGTPCVKXG ECTDQP UQWTEGU. 6hG EWTTGPV
ÿKI. 1. /QFGN HQT TGIWNCVKQP QH INWEQUG TGRTGUUGF IGPGU. 6hG RTQVGKP MKPCUG úPHþR CPF KVU TGIWNCVQTU ÿúPHôR CPF VhG DGVC-UWDWPKVU úKRþR, úKRüR, CPF )CN83Rý RNC[ C EGPVTCN TQNG KP ECVCDQNKVG TGRTGUUKQP ÿ*CTFKG GV CN. þ998ý. +P INWEQUG-ITQYP EGNNU, VhG úPHþR TGIWNCVQT[ FQOCKP CWVQKPhKDKVU VhG ECVCN[VKE FQOCKP. 6hG KPhKDKVKQP QH úPHþR NGCFU VQ C /KIþR WPFGTRhQURhQT[NCVGF VhCV OQXGU KPVQ VhG PWENGWU, YhGTG KV TGETWKVU VhG EQTGRTGUUQT úUPöRð6WRþR VQ TGRTGUU IGPG GZRTGUUKQP. ÷hGP INWEQUG NGXGN HCNNU, VhG úPHôR CEVKXCVKPI UWDWPKV DKPFU VQ VhG TGIWNCVQT[ FQOCKP CPF CWVQKPhKDKVKQP KU TGNKGXGF. #EVKXCVKQP TGSWKTGU VhG RhQURhQT[NCVKQP QH C EQPUGTXGF VhTGQPKPG TGUKFWG D[ CP WPKFGPVKHKGF RTQVGKP MKPCUG ÿ/E%CTVPG[ CPF úEhOKFV üûûþý. ïRQP CEVKXCVKQP, /KIþR DGEQOGU RhQURhQT[NCVGF D[ úPHþR CPF OQXGU QWV QH VhG PWENGWU, TGUWNVKPI KP FGTGRTGUUKQP QH INWEQUG-TGRTGUUGF IGPGU. 6hG )NE7R-4GIþR RTQVGKP RhQURhCVCUG CUUQEKCVGU YKVh VhG CEVKXCVGF EQORNGZ CPF HCEKNKVCVGU VhG VTCPUKVKQP DCEM VQ VhG KPCEVKXG UVCVG. *ZMüR TGIWNCVGU VhG RhQURhQT[NCVKQP UVCVG QH 4GIþR, YhKEh OC[ CEEQWPV KP RCTV HQT VhG GHHGEV QH *ZMüR QP úPHþR HWPEVKQP ÿúCP\ GV CN. üûûûý. ©CEh MKPCUG EQORNGZ EQPVCKPU QPG QH VhG TGNCVGF RTQVGKPU úKRþ, úKRü, QT )CN83 ÿ%CTNUQP þ999ý.
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(. 4CPFG\-)KN GV CN.
OQFGN QH TGIWNCVKQP QH IGPG GZRTGUUKQP D[ INWEQUG TGRTGUUKQP KU UhQYP KP (KIWTG þ. )NWEQUG WRVCMG KU VhG HKTUV CPF NKOKVKPI UVGR QH INWEQUG OGVCDQNKUO. +V KU ECTTKGF QWV D[ C HCOKN[ QH üû URGEKHKE hGZQUG VTCPURQTVGTU, *ZV ÿf\ECP CPF JQhPUVQP þ999ý. 6hG[ UhQY FKHHGTGPV CHHKPKVKGU HQT INWEQUG CPF CNNQY [GCUV VQ ITQY QP C DTQCF TCPIG QH INWEQUG EQPEGPVTCVKQPU. /WNVKRNG RCVhYC[U, KPENWFKPI INWEQUG TGRTGUUKQP, TGIWNCVG VhG GZRTGUUKQP QH VhG HXT IGPGU CEEQTFKPI VQ VhG COQWPV QH GZVTCEGNNWNCT INWEQUG CXCKNCDNG ÿf\ECP CPF JQhPUVQP þ999ý. 6hG UKIPCN HQT INWEQUG KPFWEVKQP QH HXT IGPG GZRTGUUKQP KU IGPGTCVGF D[ VYQ OGODGTU QH VhG INWEQUG VTCPURQTVGTU HCOKN[, úPH3R CPF 4IVüR, CEVKPI CU UGPUQTU QH VhG GZVTCEGNNWNCT INWEQUG ÿf\ECP GV CN. þ99ö; f\ECP GV CN. þ997ý. 4GEGPVN[, KV hCU DGGP UhQYP VhCV VhG KPFWEVKQP QH HXT IGPG GZRTGUUKQP D[ INWEQUG KU CNUQ GUUGPVKCN HQT VhG IGPGTCVKQP QH VhG INWEQUG TGRTGUUKQP UKIPCN ÿf\ECP üûûüý. 6hG KPVTCEGNNWNCT hGZQUGU hCXG VQ DG RhQURhQT[NCVGF VQ GPVGT KP VhG IN[EQN[VKE RCVhYC[. 6hG GZRTGUUKQP QH VhG IGPGU GPEQFKPI VhG VYQ hGZQMKPCUGU ÿHXK1 CPF HXK2ý CPF VhG INWEQMKPCUG ÿGLK1ý KU TGIWNCVGF D[ INWEQUG; DGKPI QPN[ HXK2 GZRTGUUGF YhGP INWEQUG KU WUGF CU ECTDQP UQWTEG ÿ/QTGPQ CPF *GTTGTQ üûûüý. 1VhGT IGPGU GPEQFKPI GP\[OGU QH VhG IN[EQN[VKE RCVhYC[ CTG CNUQ TGIWNCVGF D[ ECTDQP UQWTEG. PFK26 CPF PFK27 ÿGPEQFKPI RhQURhQHTWEVQMKPCUGý, PYK1 ÿR[TWXCVG MKPCUGý, ENO2 ÿGPQNCUGý, PGK1 ÿIN[EGTCVG MKPCUGý, CPF PDC1 ÿR[TWXCVG FGECTDQZ[NCUGý CTG KPFWEGF VQ XCT[KPI GZVGPF D[ VhG RTGUGPEG QH INWEQUG KP VhG OGFKWO ÿ)QPnCNXGU CPF 2NCPVC þ998ý. 3.3.1.2 The protein kinase A (PKA) pathway #FFKVKQP QH INWEQUG VQ [GCUV EGNNU ITQYP QP C RQQT ECTDQP UQWTEG VTKIIGTU C TCRKF CPF VTCPUKGPV KPETGCUG KP VhG EGNNWNCT E#/2 EQPEGPVTCVKQP ÿ6hGXGNGKP CPF FG ÷KPFG þ999ý. 6hG E#/2 KU U[PVhGUK\GF D[ VhG GP\[OG CFGP[NCVG E[ENCUG ÿGPEQFGF D[ CYR1ðCDC35ý. 6hG CEVKXKV[ QH VhKU GP\[OG KP [GCUV FGRGPFU QP VhG VYQ UOCNN )-RTQVGKPU 4CUþR CPF 4CUüR. 4GEGPVN[, VhG ) RTQVGKP-EQWRNGF TGEGRVQT U[UVGO )RTþR-)RCüR hCU DGGP KORNKECVGF KP OGFKCVKPI INWEQUG UVKOWNCVKQP QH E#/2 RTQFWEVKQP ÿ4QNNCPF GV CN. üûûüý. 6hG QPN[ VCTIGV QH E#/2 UGGOU VQ DG VhG E#/2FGRGPFGPV RTQVGKP MKPCUG, 2ò#. E#/2 DKPFU VQ VhG 2ò# TGIWNCVQT[ UWDWPKVU ÿGPEQFGF D[ BCY1ý CPF TGNGCUG VhG ECVCN[VKE UWDWPKVU ÿGPEQFGF D[ TPK1, TPK2, CPF TPK3ý. 6CTIGVU QH 2ò# KPENWFG GP\[OGU KPXQNXGF KP ECTDQP OGVCDQNKUO, NGCFKPI VQ C UVKOWNCVKQP QH HGTOGPVCVKQP ÿ4QNNCPF GV CN. üûûüý. 2ò# CNUQ EQPVTQNU VhG GZRTGUUKQP QH C NCTIG PWODGT QH IGPGU VhTQWIh VhG PGICVKXG TGIWNCVKQP QH VhG VTCPUETKRVKQP HCEVQTU /UPüR CPF /UPôR, KPENWFKPI IN[EQN[VKE GP\[OGU, GP\[OGU HQT VhG U[PVhGUKU QH IN[EGTQN, CPF TGUGTXG ECTDQh[FTCVGU ÿ©UVTWEh üûûûý. 6hG UWDEGNNWNCT NQECNK\CVKQP QH VhGUG HCEVQTU KU TGIWNCVGF D[ 2ò# CEVKXKV[. ÷hGP CEVKXG, 2ò# RhQURhQT[NCVGU VhG PWENGCT NQECNK\CVKQP UKIPCN QH /UPüR CPF KPhKDKVU KVU HWPEVKQP. úVTGUU EQPFKVKQPU FQ PQV UGGO VQ TGFWEG 2ò# CEVKXKV[ DWV VhG[ YQWNF QRGTCVG QP /UPüR D[ CHHGEVKPI C PWENGCT GZRQTV CPFðQT TGVGPVKQP OGEhCPKUO ÿ){TPGT GV CN. üûûüý. # UEhGOCVKE TGRTGUGPVCVKQP QH KPVGTCEVKQPU CPF GNGOGPVU QH VhG 2ò# RCVhYC[ KU FGRKEVGF KP (KIWTG ü.
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
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ÿKI. þ. 4GIWNCVKQP QH 2-# KP [GCUV. )NWEQUG UKIPCN KU VTCPUOKVVGF VQ VhG CFGPKNCVG E[ENCUG VhTQWIh VYQ FKHHGTGPV RCVhYC[U. E#/2 CEVKXCVGU 2ò# D[ FKUUQEKCVKQP QH VhG TGIWNCVQT[ UWDWPKVU, NGCFKPI VQ TGIWNCVKQP OCP[ GP\[OCVKE CEVKXKVKGU CPF IGPG GZRTGUUKQP. úGG VGZV HQT FGVCKNU.
3.3.1.3 Strategies to enhance fermentative performance $CMGT•U [GCUV UVTCKPU CTG EQOOGTEKCNN[ RTQFWEGF HTQO OQNCUUGU, YhGTG UWETQUG KU VhG OCLQT UWICT. 6hG DKQOCUU [KGNF QH DCMGT•U [GCUV ITQYKPI QP UWICTU KU hCORGTGF D[ KVU RTGHGTGPEG VQ ITQY D[ HGTOGPVCVKQP, GXGP KP CGTQDKE EQPFKVKQPU, YhKEh CRRGCTU VQ DG VhG EQPUGSWGPEG QH C NQY TGURKTCVQT[ ECRCEKV[. #VVGORVU VQ TGFKTGEVKQP ECTDQP HNWZGU DGVYGGP HGTOGPVCVKQP CPF TGURKTCVKQP D[ TGFWEKPI VhG CEVKXKVKGU QH HGTOGPVCVKXG GP\[OGU CTG NKOKVGF D[ VhGKT PGICVKXG GHHGEVU QP ITQYVh ÿ(NKMYGGTV GV CN. þ99ö; þ999ý. # FKHHGTGPV CRRTQCEh hCU DGGP VQ GPhCPEG TGURKTCVQT[ CEVKXKV[ D[ KPETGCUKPI VhG GZRTGUUKQP QH *CRôR. 6hKU VTCPUETKRVKQP HCEVQT KU C MG[ EQORQ-
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(. 4CPFG\-)KN GV CN.
PGPV QH VhG *CRüð3ðôð5R CEVKXCVQT EQORNGZ, TGSWKTGF HQT VhG VTCPUETKRVKQPCN KPFWEVKQP QH TGURKTCVQT[ EQORQPGPVU. 6hG QXGTRTQFWEVKQP QH HAP4 WPFGT VhG EQPVTQN QH VhG INWEQUG-KPFWEKDNG ADH1 RTQOQVGT KP C NCDQTCVQT[ UVTCKP QH S. cerevisiae NGCF VQ C OQTG QZKFCVKXG OGVCDQNKUO, TGUWNVKPI KP C UKIPKHKECPV TGFWEGF GVhCPQN RTQFWEVKQP, KORTQXGOGPV QH ITQYVh TCVG CPF C ôûù ICKP KP DKQOCUU [KGNF ÿ$NQO GV CN. üûûûý. *QYGXGT, VhG RTKOCT[ TQNG QH [GCUV KU VhG RTQFWEVKQP QH %1ü D[ CNEQhQNKE HGTOGPVCVKQP VhCV TGUWNVU KP FQWIh NGCXGPKPI. 6hGTGHQTG, CNVhQWIh [GCUV RTQFWEVKQP TGSWKTGU VhG UWRRTGUUKQP QH CNEQhQNKE HGTOGPVCVKQP, C hKIh HGTOGPVCVKXG ECRCEKV[ KU UQWIhV CV VhG GPF QH VhG KPFWUVTKCN RTQEGUU. 6Q CXQKF HGTOGPVCVKQP, DCMGT•U [GCUV RTQFWEVKQP KU RGTHQTOGF KP CGTQDKE, UWICT NKOKVGF HGF-DCEh EWNVWTGU. øCP *QGM GV CN. ÿüûûûý CPCN[\GF VhG GHHGEV QH ITQYVh EQPFKVKQPU QP HGTOGPVCVKXG ECRCEKV[ WUKPI EhGOQUVCV EWNVKXCVKQP VQ UKOWNCVG VhG KPFWUVTKCN RTQEGUU. 6hG[ HQWPF VhCV VhG TGIWNCVKQP QH HGTOGPVCVKXG ECRCEKV[ KU UVTCKP URGEKHKE. +P VhG KPFWUVTKCN [GCUV UVTCKP &úü89þþ, HGTOGPVCVKXG ECRCEKV[ KPETGCUGF NKPGCTN[ YKVh KPETGCUKPI FKNWVKQP TCVG, GXGP KP VhG TCPIG QH TCVGU VhCV RTQFWEG C HWNN[ TGURKTCVKXG ITQYVh. úQ HCT, KORTQXGOGPV QH HGTOGPVCVKXG ECRCEKV[ hCU DGGP CRRTQCEhGF GORKTKECNN[ D[ UVTCKP UGNGEVKQP RTQITCOOGU CPF D[ QRVKOK\CVKQP QH RTQEGUU RCTCOGVGTU ÿXCP *QGM GV CN. üûûûý. /QTG TGEGPVN[, FKHHGTGPV CRRTQCEhGU VQ KPETGCUG HGTOGPVCVKXG ECRCEKV[ D[ IGPGVKE GPIKPGGTKPI hCXG DGGP FGXGNQRGF. +P VhKU UGEVKQP, YG CPCN[\G VhQUG UVTCVGIKGU EQPUKUVKPI KP VhG OCPKRWNCVKQP QH S. cerevisiae QYP IGPGU. úVTCVGIKGU DCUGF KP VhG KPVTQFWEVKQP QH HQTGKIP IGPGU hCXG DGGP TGEGPVN[ FKUEWUUGF KP 2TKGVQ GV CN. ÿüûû3ý. Overexpression of glycolytic genes. ÷hGP VhG NGXGNU QH IN[EQN[VKE GP\[OGU CTG CPCN[\GF KP EQPFKVKQPU YhGTG FKHHGTGPV HGTOGPVCVKXG ECRCEKVKGU CTG QDUGTXGF, QPN[ VYQ GP\[OGU ÿRhQURhQHTWEVQMKPCUG CPF R[TWXCVG ECTDQZKNCUGý GZhKDKVGF C RQUKVKXG EQTTGNCVKQP YKVh HGTOGPVCVKXG ECRCEKV[ ÿXCP *QGM GV CN. üûûûý. 6hKU TGUWNV CTIWGU CICKPUV VhG EQPVTQN QH HGTOGPVCVKXG ECRCEKV[ D[ TGIWNCVKQP QH VhG U[PVhGUKU QH GP\[OGU VhCV EQPXGTVU KPVTCEGNNWNCT INWEQUG KP GVhCPQN. +P HCEV, VhQUG UVTCVGIKGU FGUKIPGF VQ KORTQXG VhG HGTOGPVCVKXG ECRCEKV[ QH S. cerevisiae D[ KPETGCUG QH VhG IN[EQN[VKE HNWZ VhTQWIh VhG QXGTGZRTGUUKQP QH IN[EQNKVKE IGPGU hCXG HCKNGF ÿ4CPFG\-)KN GV CN. þ999ý. 6hKU HCKNWTG UWRRQTVU VhG KFGC VhCV HNWZ EQPVTQN KU PQV GZGTVGF D[ QPN[ C UKPING GP\[OG ÿ%QTPKUh-$QYFGP CPF *QHOG[T þ995; XCP *GGUYKLM GV CN. þ999ý. ©PhCPEGOGPV QH IN[EQN[VKE HNWZ hCU DGGP QDVCKPGF D[ VhG QXGTRTQFWEVKQP QH C EQORNGVG UGV QH IN[EQN[VKE GP\[OGU, DWV QPN[ WPFGT EQPFKVKQPU QH GPhCPEGF #62 FGOCPF ÿúOKVU GV CN. üûûûý. %QPUKFGTKPI VhCV KP S. cerevisiae, VhG IN[EQN[VKE GP\[OGU CNTGCF[ EQPUVKVWVGU 3û-ö5ù QH VhG VQVCN E[VQRNCUOKE RTQVGKP EQPVGPV, VhG GPhCPEGOGPV QH VhG IN[EQN[VKE HNWZ D[ QXGTRTQFWEVKQP QH CNN VhG IN[EQN[VKE GP\[OGU KU hKIhN[ SWGUVKQPCDNG DGECWUG QH VhG GZEGUUKXG RTQVGKP DWTFGP ÿúOKVU GV CN. üûûûý. Modulation of the glycolysis/gluconegenesis balance. #P CNVGTPCVKXG VQ KORTQXKPI IN[EQN[VKE HNWZ KU OQFWNCVKQP QH VhG IN[EQN[UKU INWEQPGQIGPGUKU DCNCPEG D[ VhG KPVTQFWEVKQP QH HWVKNG E[ENGU ÿUGG CNUQ UGEV. 3.3.ü.5.ôý. 0CXCU GV CN. ÿþ993ý RNCEGF VhG IGPGU HQT VhG VYQ MG[ INWEQPGQIGPKE GP\[OGU: HTWEVQUG-þ,ö-DKURhQURhCVCUG ÿFBP1ý CPF RhQURhQGPQNR[TWXCVG ECTDQZ[MKPCUG ÿPCK1ý WPFGT VhG EQPVTQN QH VhG CNEQhQN-FGh[FTQIGPCUG-+ RTQOQVGT VQ CNNQY VhG GZRTGUUKQP QH DQVh IGPGU KP VhG RTGUGPEG QH INWEQUG. óGCUV VTCPUHQTOCPVU ECTT[KPI VhGUG EQPUVTWEVU UhQYGF CP KP-
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
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ETGCUGF UWICT-WVKNK\CVKQP TCVG ÿD[ 5ûùý, CU YGNN CU KPETGCUGF GVhCPQN HQTOCVKQP ÿD[ ôôùý CPF %1ü RTQFWEVKQP ÿD[ 35ùý KP EQORCTKUQP YKVh VhG RCTGPVCN UVTCKP. *QYGXGT, VhG[ CNUQ hCF NQPIGT IGPGTCVKQP VKOGU CPF C FGETGCUG KP DKQOCUU QH CTQWPF 3ûù ÿ0CXCU GV CN. þ993ý. Adaptation to maltose fermentation: avoiding the maltose-lag. #V VhG DGIKPPKPI QH VhG DTGCF-OCMKPI RTQEGUU, [GCUV HGTOGPVU VhG HTGG UWICTU RTGUGPV KP HNQWT ÿOCKPN[ INWEQUG, HTWEVQUG, UWETQUG, OCNVQUG CPF INWEQHTWEVCPUý. ÷hGP VhGUG UWDUVTCVGU CTG GZhCWUVGF, FQWIh HGTOGPVCVKQP EQPVKPWGU XKC FG CEVKQP QH GPFQIGPQWU ¾-CO[NCUG CPF GZQIGPQWU ½-CO[NCUG, YhKEh TGNGCUG OCNVQUG HTQO FCOCIGF UVCTEh ITCPWNGU. #NVhQWIh VhG COQWPV QH HTGG UWICTU KU NQY ÿþ-üùý, KV KU GPQWIh VQ TGRTGUU VhG U[PVhGUKU QH OCNVQUG-WVKNK\CVKQP GP\[OGU D[ ECVCDQNKVG TGRTGUUKQP ÿ)CPEGFQ þ998ý. 6hKU ECWUGU C NCI RhCUG KP %1ü RTQFWEVKQP WPVKN VhG IGPGU GPEQFKPI VhG OCNVQUG WVKNK\CVKQP RCVhYC[ CTG FGTGRTGUUGF CPF KPFWEGF ÿUGG %hCRVGT üý. 6hWU, FQWIhNGCXGPKPI CDKNKV[ EQTTGNCVGU YKVh VhG CEVKXKV[ QH VhG MAL IGPGU ÿ*KIIKPU GV CN. þ999ý, YhKEh FGVGTOKPG VhG ECRCEKV[ QH OCNVQUG VTCPURQTV CPF h[FTQN[UKU. 6hKU hCU DGGP ENGCTN[ FGOQPUVTCVGF D[ EQORCTKPI VhG OCNVCUG CPF OCNVQUG RGTOGCUG CEVKXKVKGU KP PQP-NCIIKPI ÿUNQYý XGTUWU NCIIKPI ÿTCRKFý UVTCKPU. +P VhG PQP-NCIIKPI UVTCKPU VhGUG CEVKXKVKGU YGTG TGIWNCVGF D[ INWEQUG TGRTGUUKQP, DWV YGTG CV NGCUV þü- ÿHQT OCNVCUGý CPF þ3û-HQNF hKIhGT ÿHQT OCNVCUG RGTOGCUGý KP CDUGPEG QH KPFWEKPI UWICT, CU EQPUGSWGPEG QH VhG RTGUGPEG QH URGEKHKE MALx3 TGIWNCVQT[ CNNGNGU ÿ*KIIKPU GV CN. þ999ý. úGXGTCN UVTCVGIKGU hCXG DGGP FGUKIPGF VQ CXQKF VhG NCI RGTKQF. 1UKPIC GV CN. ÿþ988ý RNCEGF VhG OCNVCUG CPF OCNVQUG RGTOGCUG IGPGU WPFGT VhG EQPVTQN QH EQPUVKVWVKXG RTQOQVGTU, KPETGCUKPI þ8 ù VhG RTQFWEVKQP QH %1ü KP RTGUGPEG QH INWEQUG. $[ WUKPI ü-FGQZK-&-INWEQUG, 4CPFG\-)KN CPF úCP\ ÿþ99ôý CPF 4KPExP GV CN. ÿüûûþý hCXG QDVCKPGF OWVCPVU VhCV UhQY FGTGIWNCVGF OCNVCUG CPF KPXGTVCUG CEVKXKVKGU. #P CNVGTPCVKXG UVTCVGI[ hCU DGGP VhG OCPKRWNCVKQP QH MG[ TGIWNCVQT[ RTQVGKPU, NKMG /KIþRð/KIüR QT *ZMüR ÿUGG (KIWTG þý. ÷hGP VhG WVKNK\CVKQP QH UWETQUG KP RTGUGPEG QH INWEQUG YCU KPXGUVKICVGF KP MIG1-FKUTWRVGF, CP CNNGXKCVKQP QH TGRTGUUKQP YCU HQWPF ÿ1NUUQP CPF 0KGNUGP üûûûý. *QYGXGT, VhGUG OQFKHKECVKQPU FKF PQV TGUWNV KP C UKIPKHKECPV EhCPIG KP DKQOCUU [KGNF ÿòNGKP GV CN. þ999ý. # TGFWEVKQP KP INWEQUG TGRTGUUKQP hCU DGGP CNUQ QDVCKPGF D[ FGNGVKQP QH HXK2. +P DCVEh EWNVWTGU, VhG hxk2 OWVCPV FKURNC[GF C HWNN[ QZKFCVKXG ITQYVh KP RTGUGPEG QH INWEQUG YKVhQWV VhG GCTN[ RTQFWEVKQP QH GVhCPQN QDUGTXGF KP VhG YKNF V[RG UVTCKP, TGUWNVKPI KP C hKIhGT DKQOCUU [KGNF ÿ&KFGTKEh GV CN. üûûþý. *QYGXGT, VhG PWNN UVTCKP GZhKDKVU C TGFWEGF ITQYVh TCVG CV hKIh INWEQUG EQPEGPVTCVKQP. +P CFFKVKQP, PQ FCVC CDQWV HGTOGPVCVKXG RGTHQTOCPEG YCU RTQXKFGF 3.3.2 ÿUOQVQNGTCPEG CPF 0Cþ VQZKEKV[ TGUKUVCPEG øCTKCVKQP KP VhG [GCUV UWTTQWPFKPI QUOQVKE RTGUUWTG KU C EQPUVCPV RhGPQOGPQP QEEWTTKPI KP CNOQUV CNN UVGRU HTQO DKQOCUU RTQFWEVKQP VQ DTGCF OCMKPI ÿ(KI. 3ý.
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(. 4CPFG\-)KN GV CN.
(KI. 3. 5QWTEGU QH QUOQVKE UVTGUU KP [GCUV RTQFWEVKQP CPF DTGCF OCMKPI.
6hGTGHQTG, [GCUVU hCXG VQ EQRG YKVh UWEEGUUKXG QUOQVKE UVTGUUHWN GPXKTQPOGPVU ÿ#VVHKGNF þ997ý, KP YhKEh VhG[ hCXG VQ ITQY CPF RGTHQTO HGTOGPVCVKQP KP VhG QRVKOCN EQPFKVKQPU TGSWKTGF HQT C IQQF DTGCF SWCNKV[. 6hWU, VhG CDKNKV[ QH [GCUV VQ YKVhUVCPF KPETGCUKPI QUOQVKE RTGUUWTGU KU C MG[ RQKPV KP VhG DTGCF OCMKPI RTQEGUU, CPF KORTQXGOGPV QH [GCUV QUOQ-VQNGTCPEG hCU DGEQOG C OCLQT HQEWU QH CVVGPVKQP FWTKPI [GCTU. 6hKU KPVGTGUV hCU KPETGCUGF D[ VhG GPQTOQWU GZRCPUKQP QH VhG UYGGV IQQFU OCTMGV, GURGEKCNN[ UKPEG VhG KPVTQFWEVKQP QH VhG OQFGTP HTQ\GP-FQWIh VGEhPQNQI[. 2TGUGPVN[, DCMGF IQQFU UWEh CU &CPKUh RCUVTKGU, FQWIhPWVU, CPF ETQKUUCPVU OCMG WR þûù QH VhG UCNGU QH DCMGF RTQFWEVU KP ©WTQRG CPF ü5-3ûù YQTNFYKFG, CPF C ITQYKPI FGOCPF KU GZRGEVGF KP VhG PGCT HWVWTG. 3.3.2.1 Sources of osmotic stress &WTKPI VhG HGF-DCVEh RTQEGUU QH DKQOCUU RTQFWEVKQP, [GCUVU CTG ITQYP KP WPFGHKPGF OGFKC QDVCKPGF DCUKECNN[ HTQO OQNCUUGU. $CVEhGU QH OQNCUUGU CTG hKIhN[ XCTKCDNG, EQPVCKPKPI FKHHGTGPV COQWPVU QH UWICTU, UCNVU, CPF QVhGT UQNWVGU. #U TGUWNV, [GCUVU CTG QHVGP GZRQUGF VQ NQY YCVGT CEVKXKV[ CPF GZEGUU QH UCNVU, [GV KP VhG OG-
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
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FKWO KP YhKEh VhG[ CTG GZRGEVGF VQ TGCEh QRVKOCN DKQOCUU [KGNFU. &QYPUVTGCO RTQEGUUKPI CNUQ KPXQNXGU h[RGT-QUOQVKE UVTGUU. 1UOQVKE UhQEM D[ CFFKPI CDQWV û.5ù QH UCNV, KU C EQOOQPN[ WUGF RTG-VTGCVOGPV KP VhG RTGRCTCVKQP QH EQORTGUUGF [GCUV DNQEMU HQT ECTTKCIG CPF UVQTCIG. 1PEG EQORTGUUGF, VhG UWTTQWPFKPI YCVGT CEVKXKV[ KU GXGP NQYGT. úVQTCIG QH [GCUV CU ETGCO QT FT[ RQYFGT CNUQ KORNKGU h[RGTQUOQVKE UVTGUU FWG VQ FGYCVGTKPI ÿ#VVHKGNF þ997ý. +P VhG DCMKPI RTQEGUU, EGNNU CTG WUWCNN[ EhCNNGPIGF VQ QUOQVKE UVTGUU D[ VhG RTGUGPEG QH UWICTU KP VhG FQWIh. 6hGUG DCMGT[ RTQFWEVU EQPVCKP WR VQ 3ûù CFFGF UWETQUG QT INWEQUGðHTWEVQUG U[TWR CU C UYGGVGPGT CIGPV, QHVGP EQODKPGF YKVh HCV, RQYFGTGF OKNM, CPF UCNV, YhKEh TGFWEGU VhG YCVGT CEVKXKV[ QH VhG FQWIh. +P HTQ\GP FQWIh, KEG HQTOCVKQP QWVUKFG VhG EGNNU HWTVhGT FGETGCUGU YCVGT CEVKXKV[, TGUWNVKPI KP CFFKVKQPCN QUOQVKE UVTGUU ÿ/[GTU CPF #VVHKGNF þ999ý. 6hG CFFKVKQP QH UCNV ÿCTQWPF üù, HNQWT DCUKUý VQ UYGGV FQWIh OC[ CNUQ ECWUG HWTVhGT GHHGEVU QP DCMKPI RGTHQTOCPEG, YhKEh CTG FKHHGTGPV HTQO VhQUG QH QUOQVKE UVTGUU. +PFGGF, CEEWOWNCVKQP QH UQFKWO CPF EhNQTKFG KP VhG E[VQUQN QH [GCUV EGNNU hCU hCTOHWN GHHGEVU HQT OQUV QH GP\[OG U[UVGOU ÿúGTTCPQ þ99öý. 6hGUG VQZKE GHHGEVU CTG QDUGTXGF CV KPVTCEGNNWNCT EQPEGPVTCVKQPU QH 0Cî CPF %N- QH 5û-þûû O/, YhKEh KU OWEh NQYGT VhCP VhG NGXGNU TGSWKTGF HQT KQPKE UVTGPIVh GHHGEVU ÿúGTTCPQ GV CN. þ997ý. úWEh KQPKE EQPVGPVU CTG GCUKN[ CEhKGXGF DGECWUG VhG NQY YCVGT CEVKXKV[ KP UYGGV FQWIh GPhCPEGU VhG GHHGEVKXG EQPEGPVTCVKQP QH DQVh KQPU. 3.3.2.2 Technological impact 6hG QUOQ-UVTGUU UWHHGTGF D[ [GCUVU KP hKIh-UWICT FQWIh ECWUGU NQUU QH HGTOGPVCVKQP ECRCEKV[, NGCFKPI VQ NQPIGT RTQQHKPI VKOGU CPF TGFWEGF RTQFWEV XQNWOG. 6Q QXGTEQOG VhGUG RTQDNGOU, DCMGTU QXGTFQUG [GCUV D[ ü-8ù ÿHNQWT DCUKUý, YhKEh KPETGCUGU RTQEGUU EQUV CPF PGICVKXGN[ CHHGEVU QTICPQNGRVKE RTQRGTVKGU. #NVGTPCVKXGN[, DCMKPI RQYFGTU, UWEh CU UQFKWO DKECTDQPCVG, ECP DG WUGF VQ GPUWTG UCVKUHCEVQT[ NGCXGPKPI. %hGOKECN NGCXGPGF RTQFWEVU FKURNC[ DCMGF NQCH YKVh C ITCKP CPF VGZVWTG TGUGODNKPI [GCUV-TCKUGF DTGCF, DWV VhG HNCXQWT KU HNCV CPF EGTGCN-NKMG ÿ4GGF CPF 0CIQFCYKVhCPC þ99þý. +P CFFKVKQP, OQFGTP EQPUWOGTU RTGHGT HQQFU YKVh HGY QT PQ CFFKVKXGU. %QPUGSWGPVN[, CP KPFWUVTKCN DCMGT•U [GCUV, YhKEh KU YGNN CFCRVGF VQ UYGGV FQWIh RTQEGUUGU, YQWNF hCXG C ITGCV KORCEV QP VhG GEQPQOKE QH VhG RTQEGUU CPF SWCNKV[ QH VhG RTQFWEVU. +P URKVG QH VhG GHHQTVU, PQPG QH VhG DCMGT•U [GCUV UVTCKPU TGRQTVGF UQ HCT EQODKPGU VhG FGUKTCDNG VGEhPQNQIKECN RTQRGTVKGU. 0GXGTVhGNGUU, DCMGT•U [GCUV CRRGCTU VQ FKURNC[ VhG CFGSWCVG OGVCDQNKE OCEhKPGT[ VQ TGURQPF CPF CFCRV VQ hKIh QUOQVKE RTGUUWTGU. 3.3.2.3 Osmotic shock: primary responses ÷hGP [GCUV EGNNU CTG UhKHVGF VQ hKIh QUOQNCTKV[, VhGTG KU C UWFFGP YCVGT NGCMCIG HTQO VhG E[VQRNCUO VQ VhG QWVUKFG ÿ(KI. ôý, FWG VQ VhG QUOQVKE ITCFKGPV HQTOGF DGVYGGP DQVh UKFGU QH VhG EGNN OGODTCPG. 6hKU FGh[FTCVKQP RTQXQMGU C NQUU QH VWTIQT, CPF ITQYVh KU KOOGFKCVGN[ CTTGUVGF. 6hGP, VYQ ENCUUGU QH TGURQPUGU CTG UVCTVGF: HKTUV, UQOG KOOGFKCVG TGUEWG OGEhCPKUOU, VhCV CTG FKTGEVGF VQ TGEQXGT YCVGT CPF VWTIQT, CXQKFKPI EGNN FGCVh, CPF UGEQPF, NQPI-VGTO UGEQPFCT[ TGURQPUGU,
7û
(. 4CPFG\-)KN GV CN.
ÿKI. ý. 4GURQPUGU VQ QUOQVKE UJQEM ÷hGP [GCUV EGNNU CTG EhCNNGPIGF VQ h[RGTQUOQVKE UVTGUU, RTKOCT[ TGUEWG OGEhCPKUOU CTG VTKIIGTGF HQT KOOGFKCVG YCVGT KPVCMG, CXQKFKPI EGNN FGCVh. úGEQPFCT[, NQPI VGTO TGURQPUGU ECWUG OGVCDQNKE TG-CTTCPIGOGPVU, YhKEh GPCDNGU VhG EGNN VQ EQORNGVGN[ CFCRV VQ VhG PGY GPXKTQPOGPV. úGG VGZV HQT FGVCKNU.
VhCV GPCDNG VhG EGNN VQ EQORNGVGN[ CFCRV KVUGNH VQ VhG PGY GPXKTQPOGPV, CPF TGUVQTG ITQYVh ÿ(KI. ôý. #DKNKV[ VQ HQTO XCEWQNGU ÿõCVVGTKEh CPF ÷CVUQP þ993ý, CPF ENQUWTG QH VhG OGODTCPG IN[EGTQN EhCPPGN, (RUþR ÿõW[VGP GV CN. þ995ý, CTG VYQ HCEVU TGNCVGF VQ KOOGFKCVG TGUEWG OGEhCPKUOU. 4GVGPVKQP QH IN[EGTQN UGGOU VQ DG C ETWEKCN UWDLGEV HQT UWTXKXCN VQ CP QUOQVKE UhQEM, DGECWUG OWVCPVU YKVh EQPUVKVWVKXG IN[EGTQN GZRQTV CTG QUOQ-UGPUKVKXG ÿ6COáU GV CN. þ999ý. )N[EGTQN KU C EQORCVKDNG QUOQN[VG, CDNG VQ EQWPVGTDCNCPEG VhG FKHHGTGPEGU KP QUOQVKE RTGUUWTG YKVhQWV KPVGTHGTG QVhGT OGVCDQNKE TGCEVKQPU, CPF KU VhG QPN[ QUOQN[VG MPQYP VQ DG CEEWOWNCVGF D[ S. cerevisiae ÿ$NQODGTI CPF #FNGT þ99üý. úGEQPFCT[, NQPI-VGTO TGUEWG OGEhCPKUOU KPENWFG de novo U[PVhGUKU CPF DCUKECNN[ UWRRQUG VTCPUETKRVKQPCN CEVKXCVKQP ÿ*QhOCPP üûûüý. 3.3.2.4 Transcriptional activation and main secondary responses 1PN[ HGY GP\[OCVKE OGEhCPKUOU CTG TGCF[ VQ IQ YhGP EGNNU CTG UWFFGPN[ GZRQUGF VQ h[RGT-QUOQVKE OGFKC. 6hG OCLQTKV[ QH GXGPVU HQNNQYKPI CP QUOQVKE UhKHV FGRGPF QP GP\[OGU VhCV hCXG VQ DG PGYN[ U[PVhGUKUGF. *GPEG, [GCUV UhQYU C RCTVKEWNCT VTCPUETKRVKQP RTQHKNG YhGP EQORCTGF VQ CP QUOQ-UVTGUUKPI GPXKTQPOGPV. 1XGT ü5ûôûû IGPGU, EQXGTKPI C YKFG XCTKGV[ QH Rh[UKQNQIKECN HWPEVKQPU UWEh CU ECTDQP CPF COKPQ-CEKF OGVCDQNKUO, TGFQZ DCNCPEG, CPVK-QZKFCPV RTQVGEVKQP, #62CUGU, OGODTCPG RTQVGKPU, EhCRGTQPGU, E[VQUMGNGVQP, CPF EGNN YCNN CTG WR-TGIWNCVGF CHVGT FKHHGTGPV EQPFKVKQPU QH QUOQVKE UhQEM ÿ)CUEh GV CN. üûûû; 4GR GV CN. üûûû; 2QUCU GV CN. üûûû;óCNG CPF $QhPGTV üûûþ; %CWUVQP GV CN. üûûþý. úVKOWNCVGF GZRTGUUKQP QH VhGUG
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
7þ
ÿKI. ü. /CKP UKIPCN VTCPUFWEVKQP RCVJYC[U TGURQPFKPI VQ QUOQVKE CPF KQPKE UVTGUU. 6hG *1) ÿHQT *KIh 1UOQNCTKV[ )N[EGTQNý RCVhYC[, QPG QH VhG HKXG /#2 ÿHQT /KVQIGP #EVKXCVGF 2TQVGKPý MKPCUG ÿ/#2òý RCVhYC[U MPQYP KP S. cerevisiae ÿ)WUVKP GV CN. þ998ý KU VhG OQUV KORQTVCPV UKIPCN VTCPUFWEVKQP OGEhCPKUO OGFKCVKPI VhG QUOQ-KPFWEGF IGPGVKE TGURQPUG. ÷hGP VhGTG KU CP KPETGCUG KP VhG GPXKTQPOGPVCN QUOQVKE RTGUUWTG, VhG UKIPCN HQT VhG IGPGVKE TGURQPUG KU VTKIIGTGF D[ VYQ KPFGRGPFGPV OGEhCPKUOU ÿ/CGFC GV CN. þ99ô; þ995ý CPF VTCPUFWEGF VhTQWIh C RhQURhQT[NCVKQP ECUECFG ÿHQT CP GZVGPUKXG TGXKGY UGG *QhOCPP üûûüý. ïRQP CEVKXCVKQP, *QIþR OKITCVGU VQ VhG PWENGWU, CPF UVCTVU VhG VTCPUETKRVKQPCN CEVKXCVKQP ÿ(GTTKIPQ GV CN. þ998; 4GKUGT GV CN. þ999ý, YhKEh CRRGCTU VQ FGRGPF, DGVYGGP QVhGTU, QP VhG VTCPUETKRVKQPCN HCEVQTU /UPüR CPF /UPôR ÿ4GR GV CN. üûûû; #NGRW\ GV CN. üûûþý. 6hG ECNEKPGWTKPð%T\þR RCVhYC[ KU CNUQ GUUGPVKCN HQT CFCRVCVKQP VQ UCNKPG UVTGUU ÿ4WUPCM CPF /GTV\ üûûûý. %CNEKPGWTKP KU CEVKXCVGF D[ %Cüî, XKC ECNOQFWNKP, C %Cüî-DKPFKPI RTQVGKP. #EVKXCVKQP QH ECNEKPGWTKP NGCFU VQ VhG FGRhQURhQT[NCVKQP QH VhG VTCPUETKRVKQPCN HCEVQT %T\þR, YhKEh KU VTCPUNQECVGF VQ VhG PWENGWU ÿ/CVhGQU GV CN. þ997; úVCVhQRQWNQU CPF %[GTV þ997ý, KPFWEKPI VhG GZRTGUUKQP QH C NCTIG UGV QH UCNV-TGURQPUKXG IGPGU ÿóQUhKOQVQ GV CN. üûûüý.
IGPGU CRRGCTU VQ DG OCKPN[ FGRGPFGPV QP VYQ YGNN EhCTCEVGTKUGF OQNGEWNCT UKIPCNNKPI RCVhYC[U ÿUGG (KIWTG 5ý, VhG *1) ÿHQT *KIh 1UOQNCTKV[ )N[EGTQNý RCVhYC[ ÿ1'4QWTMG GV CN. üûûü; *QhOCPP üûûüý, QPG QH VhG HKXG /#2 ÿHQT /KVQIGP #EVKXCVGF 2TQVGKPý MKPCUG ÿ/#2òý RCVhYC[U MPQYP KP S. cerevisiae ÿ)WUVKP GV CN. þ998ý CPF VhG ECNEKPGWTKPð%T\þR RCVhYC[ ÿ4WUPCM CPF /GTV\ üûûûý. 4GEGPVN[, C
7ü
(. 4CPFG\-)KN GV CN.
PGY /#2ò TQWVG, VhG úø) RCVhYC[ ÿHQT úVGTKNG øGIGVCVKXG )TQYVhý ÿõGG CPF ©NKQP þ999ý, UhCTKPI EQOOQP HCEVQTU YKVh VhG *1) RCVhYC[ ÿ#NQPUQ-/QPIG GV CN. üûûþý hCU CNUQ DGGP KORNKECVGF KP VhG VTCPUOKUUKQP QH VhG QUOQ-UKIPCN VQ VhG PWENGWU. 6hKU TGUWNVU KP TGRTQITCOOKPI CPF CEVKXCVKQP QH FKHHGTGPV Rh[UKQNQIKECN HWPEVKQPU CPF RTQVGEVKXG OGEhCPKUOU. /QUV QH VhGO UGGO VQ CKO OCKPN[ CV VhG RTQFWEVKQP QH QUOQRTQVGEVCPVU CPF VhG CFLWUVOGPV QH GPGTI[ OGVCDQNKUO ÿ*QhOCPP üûûüý. Osmoprotection. 6hG OCKP QUOQTGIWNCVQT[ HGCVWTG KP S. cerevisiae KU VhG CEEWOWNCVKQP QH IN[EGTQN D[ VhG CEVKQP QH IN[EGTQN-3-RhQURhCVG FGh[FTQIGPCUG GPEQFGF D[ GPD1 ÿ$NQODGTI CPF #FNGT þ99ü; õCTUUQP GV CN. þ993; #NDGTV[P GV CN. þ99ôý. 6hG RTQFWEV QH GPD1 VTCPUHQTOU FKh[FTQZ[CEGVQPGRhQURhCVG KPVQ IN[EGTQN-3RhQURhCVG, YhKEh KU VhGP EQPXGTVGF VQ IN[EGTQN D[ VhG GP\[OG IN[EGTQN-3RhQURhCVCUG, GPEQFGF D[ VhG QUOQ-CEVKXCVGF GPP2 IGPG ÿ0QTDGEM GV CN. þ99öý. )N[EGTQN KU CEEWOWNCVGF WR VQ CV NGCUV 3 hQWTU CHVGT VhG UVTGUU, CPF OWVCPVU NCEMKPI GPD1 hCXG C TGOCTMCDNG QUOQ-UGPUKVKXKV[ ÿ#NDGTV[P GV CN. þ99ôý. 1XGTCEEWOWNCVKQP QH IN[EGTQN KU CXQKFGF D[ QRGPKPI VhG IN[EGTQN EhCPPGN, (RUþR ÿõW[VGP GV CN. þ995ý, YhKEh TGIWNCVGU VhG QUOQN[VG TGVGPVKQP CPF TGNGCUG KP VhG QUOQ-CFCRVCVKQP RTQEGUU ÿúWVhGTNCPF GV CN. þ997; 6COáU GV CN. þ999ý. +P CFFKVKQP, FPS1 OQFWNCVGU VQVCN IN[EGTQN RTQFWEVKQP ÿõW[VGP GV CN. þ995ý, UWIIGUVKPI VhG GZKUVGPEG QH C HGGFDCEM OGEhCPKUO QH EQPVTQN. *QYGXGT, U[PVhGUKU QH IN[EGTQN CNUQ CHHGEVU VhG TGFQZ DCNCPEG, UKPEG VhG )RFþR CEVKXKV[ KU EQWRNGF VQ 0#&* QZKFCVKQP. 6hWU, IN[EGTQN U[PVhGUKU XKC )RFþR KU EQPUKFGTGF CU C OGVCDQNKE UKPM HQT QZKFK\KPI VhG GZEGUU QH 0#&* RTQFWEGF D[ OGVCDQNKE TGFQZ KODCNCPEGU. &TCUVKE EhCPIGU KP VhG OGVCDQNKUO FWG VQ QUOQVKE UVTGUU OC[ KPETGCUG VhGUG TGFQZ KODCNCPEGU, TGSWKTKPI HWTVhGT IN[EGTQN HQTOCVKQP QPEG VhG QUOQVKE RTGUUWTGU KPUKFG CPF QWVUKFG VhG EGNN CTG TGGSWKNKDTCVGF. Metabolic adjustment. ©ZRQUWTG QH [GCUV EGNNU VQ C hKIh QUOQVKE GPXKTQPOGPV UVQR QT UNQYFQYP VhG ITQYVh. 6hKU KU ENGCTN[ TGHNGEVGF D[ C UWFFGP FQYP-TGIWNCVKQP QH HWPEVKQPU KPXQNXGF KP EGNN RTQNKHGTCVKQP, NKMG WR-VCMG CPF CPCDQNKUO QH COKPQ CEKFU ÿ0QTDGEM CPF $NQODGTI þ998ý, PWENGQVKFG DKQU[PVhGUKU CPF &0# CPF RTQVGKP RTQFWEVKQP ÿ4GR GV CN. üûûû; 2QUCU GV CN. üûûû; óCNG CPF $QhPGTV üûûþý, GURGEKCNN[ QH TKDQUQOCN RTQVGKPU, C YGNN MPQYP OCTMGT QH EGNNWNCT ITQYVh TCVG ÿ/CIGT CPF 2NCPVC þ99þý. %QPUGSWGPVN[, EGNNU OWUV CFLWUV VhGKT GPGTIGVKE UVCVWU VQ VhG NQYGT #62 FGOCPF HQT DKQU[PVhGUKU. 4GFQZ OGVCDQNKUO PGGFU VQ DG CNUQ CFLWUVGF CEEQTFKPIN[, UKPEG DKQOCUU IGPGTCVKQP KU VhG OCKP UQWTEG QH 0#&2* GSWKXCNGPVU, C MG[ GNGOGPV HQT QZKFCVKXG RTQVGEVKQP. #U EQWNF DG GZRGEVGF, C OCKP HCEVQT VhCV EQPVTKDWVGU VQ TGFQZ CPF GPGTI[ DCNCPEKPI FWTKPI QUOQVKE CFCRVCVKQP KU VhG TGIWNCVKQP QH VhG IN[EQN[UKU, VhG EGPVTCN RCVhYC[ KP VhG OGVCDQNKUO QH S. cerevisiae. 6hWU, C ITGCV PWODGT QH IGPGU GPEQFKPI IN[EQN[VKE, KPENWFKPI FKHHGTGPV KUQGP\[OGU, CPF EQPPGEVGF RCVhYC[U GP\[OGU, CRRGCT VQ DG WR- QT FQYP-TGIWNCVGF WRQP QUOQVKE UhQEM ÿ4GR GV CN. üûûû; 2QUCU GV CN. üûûû; óCNG CPF $QhPGTV üûûþý. õCVGN[ VhGUG, KP RCTVKEWNCT VhG IN[EGTQN CPF VTGhCNQUG RCVhYC[U, hCXG DGGP RTQRQUGF VQ hCXG CP KORQTVCPV TQNG KP VhG EQPVTQN QH UWICT KPHNWZ KPVQ IN[EQN[UKU ÿ6hGXGNGKP CPF *QhOCPP þ995ý, GSWKNKDTCVKPI VhG IGPGTCVKQP CPF FGOCPF QH GPGTI[ WPFGT QUOQVKE UVTGUU ÿ$NQODGTI üûûûý.
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
73
Ionic equilibrium. # OCKP KUUWG KP VhG EGNNWNCT TGRTQITCOOKPI QH CNOQUV CNN QTICPKUOU UWDLGEVGF VQ QUOQVKE, CPF KP RCTVKEWNCT, UCNV UVTGUU KU VhG OCKPVGPCPEG QH C hKIh òîð0Cî TCVKQ ÿúNGCVQT CPF *KNN üûûþ; #RUG CPF $NWOYCNF üûûüý. +P [GCUV EGNNU, VhKU KU CEhKGXGF D[ VhG GZVTWUKQP QH 0Cî CPF VhG CEEWOWNCVKQP QH òî ÿúGTTCPQ GV CN. þ999; 4QFTKIWG\-0CXCTTQ üûûû; úGTTCPQ CPF 4QFTKIWG\-0CXCTTQ üûûþý. 6hKU KU OCKPN[ IQXGTPGF D[ VhG 2-V[RG #62CUG ©PCþR, VhCV OGFKCVGU VhG CEVKXG GHHNWZ QH 0Cî ÿ*CTQ GV CN. þ99þý, VhG UQFKWO-RTQVQP CPVKRQTVGT 0hCþR ÿ2TKQT GV CN. þ99öý, CPF VhG òî VTCPURQTVGTU 6TMþR CPF 6TMüR ÿòQ CPF )CDGT þ99þý. #PQVhGT IGPG HAL1, KU KPXQNXGF KP òî hQOGQUVCUKU, CPF KVU QXGTGZRTGUUKQP EQPHGTU UCNV, DWV PQV RWTGN[ QUOQVKE VQNGTCPEG ÿ)CZKQNC GV CN. þ99üý. ena1 FKUTWRVU CTG CNUQ UCNV UGPUKVKXG, DWV VhG[ FQ PQV UhQY QUOQUGPUKVKXKV[. #NVhQWIh QUOQVKE CPF UCNV UVTGUUGU CTG FKHHGTGPV RhGPQOGPC, DQVh TGURQPUGU WUWCNN[ QXGTNCR. ©ZRTGUUKQP QH HAL1 KU GPhCPEGF D[ QUOQVKE CPF UCNKPG UVTGUU ÿ)CZKQNC GV CN. þ99üý CPF QUOQ-VTKIIGTGF VTCPUETKRVKQPCN CEVKXCVKQP OGEhCPKUOU ÿUGG (KIWTG 5ý, YhKEh NGCFU VQ VhG KPFWEVKQP QH ENA1 ÿ/áTSWG\ GV CN. þ998ý. ïR-TGIWNCVKQP QH UCNV-TGURQPUKXG IGPGU CRRGCTU CNUQ VQ FGRGPF QP PWVTKGPVUGPUKPI U[UVGOU. 6hWU, VhG 614 RCVhYC[, C EGPVTCN EQPVTQNNGT QH EGNN ITQYVh ÿúEhOGN\NG CPF *CNN üûûû; %TGURQ CPF *CNN üûûüý CRRGCTU VQ DG KORNKECVGF, CV NGCUV KP RCTV, KP VhG GZRTGUUKQP QH ENA1 ÿ%TGURQ GV CN. üûûþý. )EPôR, C OCUVGT TGIWNCVQT QH IGPG GZRTGUUKQP KP S. cerevisiae ÿ*KPPGDWUEh CPF 0CVCTCLCP üûûüý OQFWNCVGU VhG VTCPUETKRVKQPCN CEVKXCVKQP QH HAL1 ÿ2CUEWCN-#hWKT GV CN. üûûþý. (KPCNN[, úPHþR VhG MG[ RTQVGKP MKPCUG HQT VhG ECVCDQNKVG TGRTGUUKQP RTQEGUU ÿUGG (KIWTG þý CNUQ EQPVTQNU VhG GZRTGUUKQP QH ENA1 ÿ#NGRW\ GV CN. þ997ý. 3.3.2.5 Strategies and targets for the improvement of osmotolerance &KHHGTGPV UVTCVGIKGU hCXG DGGP RTQRQUGF VQ GPFQY DCMGT•U [GCUV EGNNU YKVh GPhCPEGF ICUUKPI RGTHQTOCPEG KP hKIh-UWICT FQWIh. /QUV QH VhGO hCXG DGGP CFFTGUUGF VQ QXGTEQOG QUOQVKE CPF KQPKE KPLWT[ CPF VQ CXQKF OGVCDQNKE KODCNCPEGU. Invertase level. 6hG NGXGN QH KPXGTVCUG CEVKXKV[ hCU DGGP VTCFKVKQPCNN[ EQPUKFGTGF CP KORQTVCPV HCEVQT VhCV KPHNWGPEGU VhG DCMKPI RGTHQTOCPEG QH [GCUV KP UYGGV FQWIh ÿ©XCPU þ99ûý. 6hKU GP\[OG ENGCXGU UWETQUG, [KGNFKPI VYQ OQNGEWNGU QH UWICT RGT OQNGEWNG QH UWETQUG CPF, hGPEG, GPhCPEKPI VhG QUOQVKE RTGUUWTG. 6hGTGHQTG, DCMGT•U [GCUV UVTCKPU YKVh NQY KPXGTVCUG CEVKXKV[ ÿÉUNQYÌ UVTCKPUý hCXG DGGP UGNGEVGF HQT UYGGV FQWIh CRRNKECVKQPU. 6hGUG UVTCKPU CTG UWKVCDNG VQ GNCDQTCVG FQWIh EQPVCKPKPI NQY EQPEGPVTCVKQPU QH UWETQUG ÿ<8-þûùý, DWV RGTHQTO RQQTN[ KP NGCP FQWIh, DGECWUG QH KORCKTGF OCNVQUG CFCRVCVKQP. +P CFFKVKQP, VhGUG UVTCKPU FQ PQV UhQY KPVTKPUKE QUOQVQNGTCPEG CPF CV hKIhGT UWICT EQPVGPVU, VhG QUOQVKE RTGUUWTG KPhKDKVU VhGKT CEVKXKV[ ÿ/[GTU GV CN. þ997; *GTPCPFG\-õQRG\ GV CN. üûû3Cý. Overproduction of osmolytes. ©NGXCVGF [GCUV IN[EGTQN EQPVGPV hCU DGGP FKTGEVN[ TGNCVGF VQ QUOQ-VQNGTCPEG CPF HGTOGPVCVKQP ECRCEKV[ KP UYGGV FQWIh ÿ/[GTU GV CN. þ997; #VVHKGNF CPF òNGVUCU üûûûý. 6hGTGHQTG, FKHHGTGPV RTG-VTGCVOGPVU hCXG DGGP UWIIGUVGF VQ KUQNCVG EGNNU YKVh hKIh RQN[QN EQPVGPV HQT DCMKPI CRRNKECVKQPU. *KTCUCYC CPF óQMQKICYC ÿüûûþý TGRQTVGF C UKIPKHKECPV CEEWOWNCVKQP QH IN[EGTQN D[ RTG-UVTGUUKPI EGNNU DGHQTG VhG
7ô
(. 4CPFG\-)KN GV CN.
DTGCF OCMKPI RTQEGUU. 2TG-NQCFKPI QH EGNNU KP IN[EGTQN UQNWVKQPU CNUQ KPETGCUGU NGCXGPKPI ECRCEKV[ KP UYGGV FQWIh ÿ/[GTU GV CN. þ998ý. *QYGXGT, VhGUG OGVhQFU CTG NCDQTKQWU CPF FKHHKEWNV VQ CRRN[ KP VhG DCMKPI RTCEVKEG CPF QHVGP GZRGPUKXG. +P C FKHHGTGPV CRRTQCEh, KORTQXGOGPV QH DCMGT•U [GCUV RGTHQTOCPEG KP UYGGV FQWIh D[ OGVCDQNKE GPIKPGGTKPI QH VhG IN[EGTQN DKQU[PVhGUKU YCU RTQRQUGF ÿUGG CNUQ %hCRVGT ôý. *QYGXGT, CVVGORVU VQ KPETGCUG IN[EGTQN [KGNF CPF QUOQVQNGTCPEG KP NCDQTCVQT[ UVTCKPU D[ QXGTCEVKXCVKQP QH VhG *1) RCVhYC[ QT QXGTGZRTGUUKQP QH GPD1 HCKNGF. ©ZRTGUUKQP QH h[RGTCEVKXG *QIþR OWVCPV HQTOU KP YKNF V[RG EGNNU FTCOCVKECNN[ TGFWEGF ITQYVh TCVG WPFGT QRVKOCN ITQYVh EQPFKVKQPU CPF KPETGCUGF EGNN CIITGICVKQP CPF HNQEEWNCVKQP ÿ$GNN GV CN. üûûþý. /QTGQXGT, GPD1 QXGTGZRTGUUKQP FQGU PQV CRRGCT VQ EQPHGT QUOQVKE TGUKUVCPEG ÿ#NDGTV[P GV CN. þ99ôý. &KUTWRVKQP QH (RUþR CNQPG ÿõW[VGP GV CN. þ995; 2hKNKRU CPF *GTUMQYKV\ þ997ý QT EQODKPGF YKVh VhG QXGTGZRTGUUKQP QH GPD1 ÿõW[VGP GV CN. þ995; 6CQ GV CN. þ999ý NGCFU VQ C RTQPQWPEGF ITQYVh FGHGEV ÿ4GOK\G GV CN. üûûþý. *GPEG, CNVGTGF IN[EGTQN RTQFWEVKQP CPFðQT TGVGPVKQP CRRGCT VQ hCXG FTCOCVKE GHHGEVU KP QVhGT OGVCDQNKE DTCPEhGU, CU KV EQWNF DG GZRGEVGF HTQO VhG OWNVKRNG HWPEVKQPU QH VhG IN[EGTQN RCVhYC[. 6hKU EQWNF CNUQ GZRNCKP Yh[ hQOQNQIQWU QT hGVGTQNQIQWU GZRTGUUKQP QH QVhGT QUOQN[VGU, NKMG UQTDKVQN QT OCPKVQN, hCU NKVVNG QT PQ QUOQ-RTQVGEVKXG GHHGEVU KP S. cerevisiae ÿúhGP GV CN. þ999; #IWKNGTC CPF 2TKGVQ üûûþý. 4GTQWVKPI ECTDQP HNWZ VQ IN[EGTQN DKQU[PVhGUKU OC[ CNUQ hCXG PGICVKXG GHHGEVU QP QVhGT D[-RTQFWEVU QH CNEQhQNKE HGTOGPVCVKQP, NKMG GVhCPQN CPF GURGEKCNN[ %1ü, CU KV hCU DGGP FGOQPUVTCVGF KP YKPG [GCUV ÿ4GOK\G GV CN. þ999ý. 1XGTRTQFWEVKQP QH TGFQZ-WPEQWRNGF QUOQN[VGU OC[ RTQXKFG C OQTG UWEEGUUHWN CRRTQCEh VQ GZRCPF [GCUV ITQYVh CPF RTQFWEVKXKV[ KP UYGGV FQWIh. # ITGCV PWODGT QH EQORCVKDNG UQNWVGU, NKMG UWICTU, CNEQhQNU, UWICT-CNEQhQNU, COKPQ CEKFU, QT VhGKT FGTKXCVKXGU hCXG DGGP KFGPVKHKGF KP hCNQVQNGTCPVðhCNQRhKNKE EGNNU ÿFC %QUVC GV CN. þ998ý QRGPKPI VhG RQUUKDKNKVKGU QH PGY VCTIGVU HQT QUOQVKE RTQVGEVKQP KP [GCUV. 6hG FGGR MPQYNGFIG QH QUOQN[VG HWPEVKQP CPF TGIWNCVKQP KP QVhGT QTICPKUOU, KP RCTVKEWNCT KP RNCPVU, UWIIGUVU VYQ FTCYDCEMU VQ VhGUG CRRTQCEhGU. (KTUV, VhGTG CTG OGVCDQNKE NKOKVCVKQPU QP VhG CDUQNWVG NGXGN QH VhG VCTIGV QUOQN[VG VhCV ECP DG CEEWOWNCVGF. úGEQPF, VhG FGITGG QH KPETGCUGF VQNGTCPEG KU PQV PGEGUUCTKN[ EQTTGNCVKXG YKVh VhG COQWPVU QH QUOQRTQVGEVCPVU CVVCKPGF ÿ#RUG CPF $NWOYCNF üûûüý. Membrane protection. 6hG U[PVhGUKU QH QUOQN[VGU FQGU PQV CRRGCT VQ GZGTV C UKIPKHKECPV RTQVGEVKQP QH VhG QWVGT HCEG QH VhG EGNN OGODTCPG, YhKEh KU RGTOCPGPVN[ GZRQUGF VQ KQPKE KPLWT[ CPF QUOQVKE FCOCIG. +P RNCPVU, õ©# ÿõCVG ©ODT[QIGPGUKU-#DWPFCPVý RTQVGKPU HWPEVKQP KP EGNN RTQVGEVKQP CICKPUV YCVGT FGHKEKV KORQUGF D[ FTQWIhV, UCNKPKV[, CPF NQY VGORGTCVWTG ÿñW GV CN. þ99ö; ÷hKVUKVV GV CN. þ997ý. S. cerevisiae VTCPUHQTOCPVU GZRTGUUKPI õ©# RTQVGKPU HTQO VQOCVQ ÿ+OCK GV CN. þ99öý, YhGCV ÿúYKTG-%NCTM GV CN. þ999ý, QT DCTNG[ ÿíhCPI GV CN. üûûûý UhQYGF GPhCPEGF ITQYVh KP VhG RTGUGPEG QH C XCTKGV[ QH QUOQVKECNN[ CEVKXG EQORQWPFU, YhGTGCU PQ GHHGEV YCU QDUGTXGF KP OGFKC QH PQTOCN QUOQNCTKV[. HSP12 GPEQFGU C õ©#-NKMG RTQVGKP HTQO S. cerevisiae CPF QPG QH VhG VYQ OCLQT UOCNN hGCV UhQEM RTQVGKPU, YhKEh CEEWOWNCVGU OCUUKXGN[ WRQP VhGTOCN UVTGUU ÿ2TCGMGNV CPF /GCEQEM þ99ûý. HSP12 KU WR-TGIWNCVGF D[ QUOQVKE UVTGUU VhTQWIh VhG *1) RCVhYC[ CPF PGICVKXGN[ TGIWNCVGF D[ RTQVGKP MKPCUG # ÿøCTGNC GV CN. þ995; 4GR GV CN. þ999ý CPF INW-
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
75
EQUG TGRTGUUKQP ÿFG )TQQV GV CN. üûûûý, CPF KVU QXGTGZRTGUUKQP EQPHGTU RTQVGEVKQP CICKPUV FGUKEECVKQP CPF GVhCPQN-KPFWEGF UVTGUU ÿúCNGU GV CN. üûûûý. +OOWPQE[VQEhGOKECN CPCN[UKU KPFKECVGU VhCV *URþüR hCU C RNCUOC OGODTCPG NQECVKQP, UWIIGUVKPI C TQNG KP RTGUGTXCVKQP QH OGODTCPG UVTWEVWTG. Futile cycles. #P KORQTVCPV KUUWG KP VhG IGPGVKE KORTQXGOGPV QH KPFWUVTKCN DCMGT•U [GCUV HQT hKIh-UWICT CRRNKECVKQPU KU VhG OGVCDQNKE KODCNCPEG RTQXQMGF D[ QUOQVKE UhQEM. 1PG OGVhQF VQ CRRTQCEh VhKU RTQDNGO hCU DGGP VhG GPIKPGGTKPI QH HWVKNG E[ENGU ÿ4QIGTU CPF ú\QUVCM þ987; 0CXCU GV CN. þ993ý, YhKEh YCU HWTVhGT GZRNQTGF CPF ENCKOGF D[ )KUV-$TQECFGU KP C RCVGPV CRRNKECVKQP ÿXCP 4QQKLGP GV CN. þ99ôý. ©PIKPGGTGF UVTCKPU QXGTGZRTGUUKPI VhG INWEQPGQIGPKE FBP1 CPF PCK1 IGPGU YGTG CDNG VQ RTQFWEG þ9ù OQTG ICU EQORCTGF YKVh VhG RCTGPVCN UVTCKP. 0GXGTVhGNGUU, C UKIPKHKECPV TGFWEVKQP QP DKQOCUU [KGNF YCU QDUGTXGF ÿUGG CNUQ UGEV. 3.3.þ.3ý. /QTG TGEGPVN[, VYQ ü-FGQZ[-&-INWEQUG-TGUKUVCPV OWVCPVU QH DCMGT•U [GCUV hCXG DGGP UhQYP VQ FKURNC[ KORTQXGF HGTOGPVCVKXG ECRCEKV[ QP UYGGV FQWIh ÿ4KPExP GV CN. üûûþý. #NVhQWIh PQ QDXKQWU GZRNCPCVKQP QH VhKU RhGPQV[RG YCU RTQXKFGF, VhG VYQ OWVCPVU GZhKDKVGF GPhCPEGF ü-FGQZ[INWEQUG-ö-RhQURhCVG ÿü-&1)ý RhQURhCVCUG CEVKXKV[, GPEQFGF D[ VhG hQOQNQIQWU IGPGU DOG1 CPF DOG2 ÿúCP\ GV CN. þ99ô; 4CPFG\-)KN GV CN. þ995ý. +P CFFKVKQP VQ ü-&1), DQVh RTQVGKPU UhQY C UKIPKHKECPV CEVKXKV[ CICKPUV INWEQUG-ö-RhQURhCVG, CPF QVhGT UWICT RhQURhCVGU ÿ4CPFG\-)KN GV CN. þ995ý, CPF VhGTGHQTG, KPETGCUGF NGXGNU QH RhQURhCVCUG EQWNF IGPGTCVG C HWVKNG E[ENG YKVh PGV h[FTQN[UKU QH #62. +P EQPUQPCPEG YKVh VhKU, DOG2 hCU DGGP TGRQTVGF VQ DG QUOQVKECNN[ KPFWEGF CPF *1)-RCVhYC[ FGRGPFGPV ÿ6UWLKOQVQ GV CN. üûûûý UWIIGUVKPI C HWPEVKQPCN TQNG QH VhKU IGPG KP QUOQRTQVGEVKQP. High K+/Na+ ratio. 6hG CFFKVKQP QH UCNV VQ hKIh-UWICT FQWIh hCU hCTOHWN GHHGEVU QP [GCUV RGTHQTOCPEG ÿ*GTPCPFG\-õQRG\ GV CN. üûû3Cý. %QPUGSWGPVN[, 0Cî TGUKUVCPEG UhQWNF DG EQPUKFGTGF KP VhG EQODKPCVKQP QH VTCKVU VhCV UhQWNF GZhKDKV C EQOOGTEKCN DCMGT•U [GCUV UVTCKP. úVWFKGU KP [GCUV hCXG EQXGTGF DCUKE OGEhCPKUOU QH KQP hQOGQUVCUKU CPF hCXG KFGPVKHKGF MG[ IGPGU KP VhG OCKPVGPCPEG QH C hKIh òîð0Cî TCVKQ ÿúGTTCPQ GV CN. þ999; 4QFTKIWG\-0CXCTTQ üûûûý. $GVYGGP VhGO, IGPGU KPXQNXGF KP GZVTWUKQP CPF XCEWQNCT EQORCTVOGPVCVKQP QH 0Cî KQPU CRRGCT CU HWPFCOGPVCN. ©PIKPGGTKPI QH VhGUG DCUKE OGEhCPKUOU YKNN CNNQY VhG IGPGTCVKQP QH 0Cî-VQNGTCPV DCMGT•U [GCUV. 4GEGPVN[, RTQVGKP MKPCUGU CPF UKIPCNNKPI RCVhYC[U KPXQNXGF KP VhG UCNV TGURQPUG hCXG DGGP CNUQ KFGPVKHKGF CPF EhCTCEVGTKUGF ÿúGTTCPQ CPF 4QFTKIWG\-0CXCTTQ üûûþý. /WVCVKQPU KP VhGUG TGIWNCVQT[ GNGOGPVU hCXG FTCOCVKE GHHGEVU QP UCNV UGPUKVKXKV[, CPF VhGTGHQTG, VhG[ CTG QDXKQWU ECPFKFCVGU HQT [GCUV OCPKRWNCVKQP. 3.3.3 %T[QTGUKUVCPEG +P VhG DCMKPI KPFWUVT[, HTQ\GP-FQWIh VGEhPQNQI[ hCU DGGP CEEGRVGF FWG VQ UGXGTCN CFXCPVCIGU, YhKEh KPENWFG UWRRN[KPI QXGP-HTGUh DCMGT[ RTQFWEVU VQ EQPUWOGTU CPF KORTQXKPI NCDQWT EQPFKVKQPU HQT DCMGTU. 1XGT VhG NCUV FGECFGU, VhG SWCNKV[ QH DTGCF CPF RCUVT[ QDVCKPGF HTQO HTQ\GP FQWIh hCU DGGP OCTMGFN[ KORTQXGF D[ CFCRVCVKQP QH PQXGN DTGCF OCMKPI VGEhPQNQI[ ÿ÷CPI CPF 2QPVG þ995; +PQWG GV CN.
7ö
(. 4CPFG\-)KN GV CN.
þ995ý. %WTTGPVN[, TGEKRGU KPENWFKPI PGY KPITGFKGPVU CPF CFFKVKXGU, URGEKHKE GSWKROGPV HQT TCRKF CPF NQY-VGORGTCVWTG OKZKPI, QT FGGR HTGG\KPI VWPPGNU CTG CXCKNCDNG KP VhG NCTIG KPFWUVT[. %QORCTGF VQ VhKU, VhG UGNGEVKQP CPF KPVTQFWEVKQP KP VhG DTGCF OCMKPI RTQEGUU QH PGY ET[QTGUKUVCPV UVTCKPU hCU DGGP SWKVG NKOKVGF. 1P VQR QH VhKU, NKVVNG KU MPQYP CDQWV VhG Rh[UKQNQIKECN HCEVQTU VhCV OCMG [GCUV OQTG HTGG\GTGUKUVCPV CPF VhG OQNGEWNCT OGEhCPKUOU KPXQNXGF KP ET[QTGUKUVCPEG TGOCKP WPENGCT. #EEQTFKPIN[, VhG NQUU QH XKCDKNKV[ CPF NGCXGPKPI ECRCEKV[ QH DCMGT•U [GCUV FWTKPI HTGG\KPI CPF HTQ\GP UVQTCIG KU VhG OCKP HCEVQT VhCV NKOKVU VhG GZRCPUKQP CPF FGXGNQROGPV QH VhGUG RTQFWEVU ÿ)oNKPCU GV CN. þ993; 4CPFG\-)KN GV CN. þ999ý. 3.3.3.1 Effects of freezing and frozen storage 6hG HKTUV UVTGUU GPEQWPVGTGF D[ DCMGT•U [GCUV EGNNU FWTKPI RTGRCTCVKQP QH HTQ\GP FQWIh KU C UhCTR FGETGCUG, CHVGT OKZKPI, KP VhG UWTTQWPFKPI VGORGTCVWTG, C OQFKHKECVKQP MPQYP CU EQNF-UhQEM. 6hKU GPXKTQPOGPV CNVGTCVKQP KORCKTU VhG RTQRGT HWPEVKQPKPI QH DQVh VhG OGODTCPG CPF VhG VTCPUNCVKQPCN CRRCTCVWU CU TGUWNV QH TGFWEGF OGODTCPG HNWKFKV[ CPF UVCDKNK\CVKQP QH VhG UGEQPFCT[ UVTWEVWTGU QH &0# CPF 40# ÿ6hKGTKPIGT GV CN. þ998; +PQW[G þ999ý. ÷hGTGCU RQUKVKXG EQNF VGORGTCVWTGU NGCF VQ VhG U[PVhGUKU QH URGEKHKE RTQVGKPU CUUQEKCVGF YKVh VhG FGXGNQROGPV QH VTCPUKGPV RhGPQV[RKE CFCRVCVKQP ÿúChCTC GV CN. üûûüý, HTGG\KPI KU QHVGP C NGVhCN UVTGUU VQ EGNNU. #V UWD\GTQ VGORGTCVWTGU, VhG FGNGVGTKQWU GHHGEVU QP [GCUV EGNNU FGRGPF QH VhG HTGG\KPI TCVG. #V hKIh HTGG\KPI TCVGU, EGNNU CTG KPLWTGF D[ VhG HQTOCVKQP QH KPVTCEGNNWNCT KEG ET[UVCNU, NGCFKPI VQ OCETQOQNGEWNGU CPF OGODTCPG FGPCVWTCVKQP ÿ/QTTKU GV CN. þ988ý. ïNVTCUVTWEVWTCN GZCOKPCVKQP QH VhGUG EGNNU UhQYU FKUEQPVKPWQWU PWENGCT OGODTCPGU, FKUCRRGCTCPEG QH XCEWQNGU CPF &0# URTGCFU CNN QXGT VhG EGNNU ÿòCWN GV CN. þ99üý. +P EQPVTCUV, CV NQY HTGG\KPI TCVG, QUOQVKE UhTKPMCIG QH VhG EGNN CPF HTQ\GP GZVTCEGNNWNCT YCVGT KU QDUGTXGF. +P VhKU ECUG, VhG EGNNU DGEQOG GZRQUGF VQ h[RGTQUOQVKE UQNWVKQPU CPF GSWKNKDTCVG D[ OQXGOGPV QH YCVGT CETQUU VhG OGODTCPGU ÿ÷QNHG CPF $T[CPV þ999ý. &WTKPI HTQ\GP UVQTCIG, KEG ET[UVCN ITQYVh ECP HWTVhGT FGVGTKQTCVG VhG RNCUOC OGODTCPG CPF KORCKT CEVKXKV[ KP FKHHGTGPV EGNNWNCT U[UVGOU. (KPCNN[, FWTKPI VhG VhCYKPI RTQEGUU, EGNNU ECP UWHHGT DKQEhGOKECN FCOCIG D[ QZKFCVKXG UVTGUU ÿ*GTOGU-õKOC CPF úVQTG[ þ993ý. 1XGTCNN, VhGUG QDUGTXCVKQPU KPFKECVG VhCV HTGG\KPI KU C EQORNGZ CPF OWNVKHCEGVGF UVTGUU, KP YhKEh FKHHGTGPV UVTGUUQTU CPF UVTGUU TGURQPUGU CRRGCT VQ RNC[ KORQTVCPV TQNGU. +V KU PQV UWTRTKUKPI; VhGTGHQTG, VhCV HTGG\KPI VQNGTCPEG KU NKMGN[ VQ KPXQNXG FKHHGTGPV OGEhCPKUOU YQTMKPI KP EQPEGTV. 3.3.3.2 Cellular factors involved in cryoresistance $CMGT•U [GCUV UVTCKPU YKVh KORTQXGF ET[QTGUKUVCPEG hCXG DGGP UGNGEVGF HTQO PCVWTCN UQWTEGU CPF [GCUV EWNVWTG EQNNGEVKQPU D[ VhGKT CDKNKV[ VQ UWTXKXG UWEEGUUKXG HTGG\KPI-VhCYKPI E[ENGU ÿ1FC CPF 6QPQOWTC þ993; #NOGKFC CPF 2CKU þ99öCý QT hCXG DGGP EQPUVTWEVGF D[ h[DTKFKUCVKQP ÿ0CMCICYC CPF 1WEhK þ99ô; %QFxP GV CN. üûû3ý QT OWVCVKQP ÿ6CMCIK GV CN. þ997; XCP &KLEM GV CN. üûûû; 6GWPKUUGP GV CN. üûûüý. 6hG Rh[UKQNQIKECN UVWF[ QH VhGUG UVTCKPU hCU NGF VQ VhG EQTTGNCVKQP QH ET[QTGUKUVCPEG YKVh XCTKQWU EGNNWNCT HCEVQTU.
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
77
Respiratory growth. óGCUV EGNNU UhQY XCTKQWU FGITGGU QH VQNGTCPEG VQ UVTGUUGU FGRGPFKPI QP VhGKT Rh[UKQNQIKECN UVCVG CPF ITQYVh EQPFKVKQPU ÿ/CIGT CPF (GTTGKTC þ993; ÷GTPGT-÷CUhDWTPG GV CN. þ993ý. +V KU IGPGTCNN[ CUUWOGF VhCV EGNNU KP UVCVKQPCT[ RhCUG CTG OQTG TGUKUVCPV VQ XCTKQWU UVTGUUGU VhCP CEVKXGN[ INWEQUG-ITQYKPI EGNNU. +P VhG ECUG QH HTGG\G UVTGUU, OCZKOWO VQNGTCPEG QEEWTU PQV FWTKPI UVCVKQPCT[ RhCUG, DWV FWTKPI TGURKTCVQT[ ITQYVh QP GVhCPQN ÿõGYKU GV CN. þ993; 2CTM GV CN. þ997ý. +P CFFKVKQP, TGURKTCVQT[-FGHKEKGPV OWVCPVU CTG OQTG UGPUKVKXG VQ HTGG\G-VhCY UVTGUU, RTQDCDN[ GZRNCKPGF D[ VhG QZKFCVKXG DWTUV FWTKPI VhCYKPI ÿ*GTOGU-õKOC CPF úVQTG[ þ993ý. úWEh RTQEGUU YQWNF NGCF VQ VhG IGPGTCVKQP QH TGCEVKXG QZ[IGP URGEKGU ÿ41úý YKVh EQPUGSWGPV QZKFCVKXG FCOCIG VQ EGNNWNCT EQORQPGPVU ÿøCNGPVKPG GV CN. þ998ý. 6hG OKVQEhQPFTKCN TGURKTCVQT[ EhCKP RTQXKFGU C RTKOCT[ UQWTEG QH UWRGTQZKFG ÿíhCPI GV CN. þ998ý, CPF hGPEG, CEVKXGN[ TGURKTKPI OKVQEhQPFTKC UWRRN[ C HKTUV NKPG QH FGHGPEG CICKPUV 41ú ÿ2CTM GV CN. þ998; /QTCFCU-(GTTGKTC CPF %QUVC üûûûý. Trehalose. /WEh QH VhG FCOCIG VQ EGNNU KP C HTGG\G-VhCY UVTGUU TGUWNVU HTQO HTGG\KPI TCVhGT VhCP HTQO VhCYKPI CPF KU FWG RTKOCTKN[ VQ Rh[UKECN HCEVQTU UWEh CU KEG ET[UVCN HQTOCVKQP CPF FGh[FTCVKQP ÿ2CTM GV CN. þ997ý. 6hG GHHGEV QH VTGhCNQUG QP KEG PWENGCVKQP KU WPENGCT, DWV KVU TQNG CU C YCVGT UWDUVKVWVG FWTKPI FGh[FTCVKQP KU YGNN FQEWOGPVGF ÿúCPQ GV CN. þ999ý. 6hWU, RTQRCICVKQP EQPFKVKQPU VhCV NGF VQ VTGhCNQUG QXGTCEEWOWNCVKQP hCXG DGGP WUGF D[ DCMGT•U [GCUV RTQFWEGTU HQT [GCTU VQ CXQKF CWVQN[UKU CPF KPETGCUG NGCXGPKPI ECRCEKV[. *KIh NGXGNU QH VTGhCNQUG hCXG CNUQ DGGP EQTTGNCVGF YKVh TGUKUVCPEG VQ HTGG\KPI CPF GVhCPQN UhQEM ÿXCP &KLEM GV CN. þ995; òKO GV CN. þ99ö; õGYKU GV CN. þ997ý. %GNNWNCT NGXGNU QH VTGhCNQUG CTG EQPVTQNNGF D[ C DCNCPEG DGVYGGP U[PVhGUKU CPF FGITCFCVKQP. +P NCDQTCVQT[ [GCUV UVTCKPU, FKUTWRVKQP QH CEKFKE XCEWQNCT VTGhCNCUG ÿATH1ý KORTQXGU EGNN UWTXKXCN CHVGT HTGG\G-VhCY, CPF VhKU GHHGEV EQTTGNCVGU YKVh ITGCVGT NGXGNU QH EGNNWNCT VTGhCNQUG ÿòKO GV CN. þ99öý. *QYGXGT, VhKU GHHGEV YCU UVTCKP FGRGPFGPV YhGP PGWVTCN E[VQUQNKE VTGhCNCUG NTH1 CEVKXKV[ YCU CDQNKUhGF ÿXCP &KLEM GV CN. þ995; òKO GV CN. þ99öý. nth1 OWVCPVU HTQO EQOOGTEKCN UVTCKPU UhQYGF GPhCPEGF ET[QVQNGTCPEG CPF VhKU GHHGEV YCU CUUQEKCVGF YKVh C hKIh KPVTCEGNNWNCT VTGhCNQUG NGXGN ÿúhKOC GV CN. þ999ý. +P HWNN CITGGOGPV YKVh VhKU, VTGhCNQUG RTG-NQCFGF DCMGT•U [GCUV EGNNU GZhKDKVGF C ENGCT KORTQXGOGPV KP HTQ\GP FQWIh NGCXGPKPI ÿ*KTCUCYC GV CN. üûûþý. Accumulation of amino acids. +P [GCUV, NKMG KP XCTKQWU QVhGT QTICPKUOU, CEEWOWNCVKQP QH EhCTIGF COKPQ CEKFU, UWEh CU INWVCOCVG, N[UKPG, CPF CTIKPKPG, TGUWNVU KP hKIhGT HTGG\KPI VQNGTCPEG ÿ6CMCIK GV CN. þ997ý. 6hGUG EQORQWPFU YQWNF hCXG C HWPEVKQP QH KQP-EQCVKPI QP VhG UWTHCEG QH OGODTCPG EQORQPGPVU CPF RTQVGKPU RTGXGPVKPI VhGKT FGPCVWTCVKQP. 6hG FGITGG QH ET[QRTQVGEVKQP OC[ FGRGPF QP OGODTCPG UVTWEVWTG EhCTCEVGTKUVKEU QH GCEh UVTCKP. 6Q ENCTKH[ VhG ET[QRTQVGEVKXG TQNG QH EhCTIGF COKPQ CEKFU KP EQOOGTEKCN DCMGT•U [GCUV, C hQOQ\[IQWU FKRNQKF CTIKPCUGFGHKEKGPV OWVCPV hCU DGGP EQPUVTWEVGF D[ FKUTWRVKQP QH CAR1 ÿúhKOC GV CN. üûû3ý. /QNCUUGU-ITQYP EGNNU QH VhKU UVTCKP CEEWOWNCVGF hKIhGT NGXGNU QH INWVCOCVG CPF
78
(. 4CPFG\-)KN GV CN.
ÿKI. û. 5EJGOCVKE TGRTGUGPVCVKQP QH VJG LúRTQNKPG DKQU[PVJGVKE RCVJYC[. (WNN CTTQYU KPFKECVG FKTGEV OGVCDQNKE UVGRU. 2TQþR: Ä-)NWVCO[N MKPCUG; 2TQüR: Ä-)NWVCO[N RhQURhCVG TGFWEVCUG; 2WVþR: 2TQNKPG QZKFCUG; 2WVüR: •‰-2[TTQNKPG-5-ECTDQZ[NCVG FGh[FTQIGPCUG; 2TQ3R: •‰-2[TTQNKPG-5-ECTDQZ[NCVG TGFWEVCUG; %CTþR: #TIKPCUG; %CTüR: 1TPKVhKPG VTCPUCOKPCUG.
UhQYGF þû VQ þûûû VKOGU hKIhGT UWTXKXCN TCVG WRQP UGXGTCN E[ENGU QH HTGG\GðVhCY. ©PhCPEGF NGCXGPKPI CEVKXKV[ ÿüûùý YCU CNUQ QDUGTXGF FWTKPI HTQ\GP-FQWIh UVQTCIG. 6hG ET[QRTQVGEVKXG GHHGEV QH RTQNKPG ECP DG RTGUWOGF VQ DG WPKXGTUCN, DGECWUG KV KU VhQWIh VhCV NKMG VTGhCNQUG, RTGUGTXGU OGODTCPG UVTWEVWTG FWTKPI HTGG\KPI, CEVKPI CU C RTQVGKP HQNFKPI EhCRGTQPG ÿúCOWGN GV CN. üûûûý. +P S. cerevisiae, EGNNU hCTDQWTKPI C OWVCVGF PRO1 IGPG, YhKEh GNGXCVGF VhG CEVKXKV[ QH Ä-INWVCO[N MKPCUG ÿPRO1ý CPF Ä-INWVCO[N RhQURhCVG TGFWEVCUG ÿPRO2ý, DQVh KPXQNXGF KP RTQNKPG DKQU[PVhGUKU HTQO INWVCOCVG ÿUGG (KIWTG öý, CEEWOWNCVGF RTQNKPG CPF UhQYGF HTGG\KPI VQNGTCPEG CV UhQTV UVQTCIG VKOG ÿ6CMCIK GV CN. þ997; /QTKVC GV CN. üûû3ý. úKOKNCT TGUWNVU YGTG QDVCKPGF YhGP C PUT1-FKUTWRVGF OWVCPV, FGHKEKGPV KP RTQNKPG QZKFCUG ÿ(KI. öý, YCU EQPUVTWEVGF ÿ6CMCIK GV CN. üûûûý. Lipid composition. (NWKFKV[ QH VhG OGODTCPG KU KORQTVCPV HQT OCKPVCKPKPI VhG DCTTKGT RTQRGTVKGU QH VhG NKRKF DKNC[GT CPF HQT VhG CEVKXCVKQP CPF HWPEVKQP QH EGTVCKP OGODTCPG-DQWPF GP\[OGU. # WPKXGTUCN TGURQPUG VQ NQY VGORGTCVWTG KU C EhCPIG KP VhG NKRKF EQORQUKVKQP QH OGODTCPGU KP QTFGT VQ OCKPVCKP VhG QRVKOCN HNWKFKV[.
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
79
4GEGPV UVWFKGU UWIIGUV VhCV EhCPIGU KP UQOG NKRKF HTCEVKQPU EQWNF CNUQ DG KPXQNXGF KP HTGG\KPI VQNGTCPEG CESWKUKVKQP. 6hWU, EQORCTCVKXG CPCN[UKU QH VhG NKRKF EQORQUKVKQP QH EQOOGTEKCN DCMGT•U [GCUV, UhQYKPI FKHHGTGPV FGITGGU QH HTGG\KPIUGPUKVKXKV[, TGXGCNGF C RQUKVKXG EQTTGNCVKQP DGVYGGP VhG TCVKQ UVGTQN VQ RhQURhQNKRKF QH VhG RNCUOC OGODTCPG CPF ET[QVQNGTCPEG. 1P VhG EQPVTCT[, VQVCN NKRKF CPF UVGTQN EQPVGPV, CU YGNN CU VhG RhQURhQNKRKF WPUCVWTCVKQP KPFGZ, FKF PQV EQTTGNCVG YKVh DCMGT•U [GCUV ET[QVQNGTCPEG ÿ)oNKPCU GV CN. þ99þ; /WTCMCOK GV CN. þ99öý. 3.3.3.3 The cold and freeze response #FCRVCVKQP QH [GCUV EGNNU VQ C FQYPUhKHV KP VGORGTCVWTG QEEWTU D[ CNVGTCVKQPU KP OGVCDQNKE TCVG, KQP EQPEGPVTCVKQP, OGODTCPG EQORQUKVKQP, CPF D[ EhCPIG KP IGPG GZRTGUUKQP. %QORCTGF YKVh VhG hWIG COQWPV QH KPHQTOCVKQP EQPEGTPKPI VhG EQNF CPF HTGG\G TGURQPUG KP RNCPVU CPF DCEVGTKC, VhGUG UVTGUUGU hCXG TGEGKXGF NKVVNG CVVGPVKQP KP S. cerevisiae. ïPNKMG QVhGT GWMCT[QVGU, RTGXKQWU UVWFKGU D[ 2CTM GV CN. ÿþ997ý UhQYGF VhCV S. cerevisiae FQGU PQV GZRGTKGPEG RhGPQV[RKE CFCRVCVKQP VQ ET[QVQNGTCPEG. 6hWU, NGVhCN GHHGEVU KP [GCUV ECWUGF D[ HTGG\KPI YGTG PQV RTGXGPVGF QT CNNGXKCVGF GKVhGT D[ EQNF-UhQEM RTG-VTGCVOGPV QT D[ TGRGCVGF HTGG\GðVhCY E[ENGU. *QYGXGT, TGEGPVN[ KUQNCVGF HTGG\G VQNGTCPV OWVCPVU, YGTG QDVCKPGF D[ C RTQEGFWTG VhCV EQODKPGU ïø OWVCIGPGUKU CPF üûû TQWPFU QH HTGG\KPIðVhCYKPI ÿ6GWPKUUGP GV CN. üûûüý. 4GEGPV GXKFGPEGU KP QWT NCDQTCVQT[ CNUQ KPFKECVG VhCV EGNNU QH KPFWUVTKCN UVTCKPU ITQYKPI CEVKXGN[ CV þ5·% FKURNC[ GPhCPEGF HTGG\G CPF HTQ\GP-UVQTCIG TGUKUVCPEG VhCP VhQUG ITQYP CV 3û·% ÿ4QFTKIWG\-øCTICU ú, 4CPFG\-)KN (, WPRWDNKUhGF TGUWNVUý. 6hWU, C YGNN CFCRVGF OGVCDQNKUO CV NQY VGORGTCVWTGU CRRGCTU VQ KPFWEG hKIhGT TGUKUVCPEG VQ C NGVhCN HTGG\G UVTGUU KP S. cerevisiae, CU KV hCU DGGP FGOQPUVTCVGF HQT QVhGT [GCUVU ÿ6hCOOCXQPIU GV CN. üûûûý. +V KU VhGTGHQTG YQTVhYhKNG VQ TGXKUKV VhG IGPGU TGRQTVGF CU EQNF-KPFWEGF QT GUUGPVKCN HQT ITQYVh CV EQNF VGORGTCVWTGU. Laboratory strains. 2TGXKQWU YQTM hCU NGF VQ VhG KFGPVKHKECVKQP D[ FKHHGTGPVKCN UETGGPKPI QH UGXGTCN EQNF-KPFWEGF IGPGU, TIP1 ÿVGORGTCVWTG-KPFWEGF RTQVGKPý CPF VhTGG hQOQNQIWGU, TIR1, TIR2, CPF TIR4, YhKEh GPEQFG RTQVGKPU EQPVCKPKPI C UGTKPG-TKEh %-VGTOKPCN FQOCKP ÿòQPFQ CPF +PQW[G þ99þý. *QYGXGT, VhGKT FKUTWRVKQP FKF PQV UhQY CP[ CRRCTGPV RhGPQV[RG CV NQY VGORGTCVWTGU ÿòQYCNUMK GV CN. þ995ý. #PQVhGT UVWF[ D[ íhCPI GV CN. ÿüûûþý RTQXKFGF CFFKVKQPCN KPHQTOCVKQP QP IGPGU WR-TGIWNCVGF D[ C VGORGTCVWTG FGETGCUG HTQO 3û VQ þû·%. 6hG NKUV KPENWFGU YKL183w, YLR011w, FBA1, GPEQFKPI C HTWEVQUG DKRhQURhCVG CNFQNCUG, RPL2B, YhKEh GPEQFGU VhG öûú TKDQUQOCN UWDWPKV RTQVGKP CPF NOP1, C PWENGCT RTQVGKP KPXQNXGF KP TKDQUQOG DKQIGPGUKU. # EQNF-UGPUKVKXG ITQYVh RhGPQV[RG QH rpl2b YCU QDUGTXGF. 4GEGPVN[, INQDCN IGPG GZRTGUUKQP UVWFKGU hCXG CNUQ DGGP EQPFWEVGF QP EQNFUhQEMGF EGNNU, EQPHKTOKPI VhG KPFWEVKQP CPF TGRTGUUKQP QH RTGXKQWUN[ TGRQTVGF IGPGU ÿ)CUEh GV CN. üûûûý. #P GZVGPUKXG CPCN[UKU QH IGPG RTQHKNG EhCPIG CNQPI C EQNF UVTGUU VTGCVOGPV UhQYGF VhCV 93ô IGPGU YGTG WR-TGIWNCVGF YhKNG 75ö YGTG FQYP-TGIWNCVGF FWTKPI 8 h QH GZRQUKVKQP CV þû·% ÿúChCTC GV CN. üûûüý. (WPEVKQPCN CPF ENWUVGTKPI CPCN[UKU QH VGORQTCN EhCPIGU UhQYGF VhCV IGPGU TGNCVGF VQ T40#
8û
(. 4CPFG\-)KN GV CN.
U[UPVhGUKU, UVTGUU RTQVGEVKQP, CPF TKDQUQOCN RTQVGKPU YGTG WR-TGIWNCVGF KP C VKOGFGRGPFGPVN[ HCUhKQP. 6hG KORQTVCPEG QH VhGUG HWPEVKQPU KP EQNF CFCRVCVKQP hCU DGGP HWTVhGT EQPHKTOGF D[ U[UVGOCVKE CPCN[UKU QH IGPG FKUTWRVKQPU. 6CDNG ü IKXGU CP QXGTXKGY QH UQOG XGTKHKGF EQNF-UGPUKVKXG OWVCPVU D[ FKHHGTGPV ITQWRU. 6hKU UhQYU GXKFGPEG QH VhG TGSWKTGOGPV QH C TGOQFGNNKPI QH VhG VTCPUNCVKQPCN OCEhKPGT[ CPF VhG UGEQPFCT[ UVTWEVWTG QH PWENGKE CEKFU EQORTQOKUGF D[ EQNF-UhQEM. # EQORNGVG NKUV QH VhG þûö 14(U KPENWFGF KP VhG ECVCNQIWG QH EQNF-UGPUKVKXKV[ ÿ*CORUG[ þ997ý ECP DG HQWPF CV VhG HQNNQYKPI ï4õ, hVVR:ððOKRU.IUH.FGðRTQLð [GCUVð%ó)&ðFDðKPFGZ.hVON. 6CDNG þ. 5QOG GZCORNGU QH S. cerevisiae EQNFúUGPUKVKXG OWVCPVU ÿWPEVKQPCN ECVGIQT[ 2TQVGKP U[PVhGUKU 6TCPUETKRVKQP
5WDúECVGIQT[ùHWPEVKQP
4KDQUQOG DKQIGPGUKU 4KDQUQOCN RTQVGKPU 4KDQUQOG EhCRGTQPG *ú27û HCOKN[ T40# RTQEGUUKPI 40#-FGRGPFGPV #62CUG *GNKECUG
øGPG PCOG
RPS11A, RPS11B, ASC1 SSB1, SSB2 MPP10 DRS1
O40# RTQEGUUKPI, URNKEKPI 40#-hGNKECUG TGNCVGF RTQVGKP 40#-FGRGPFGPV #62CUG *GNKECUG O40# U[PVhGUKU ÿ%6&ý MKPCUG UWDWPKVU %GNNWNCT VTCPU- ©ZVTCEGNNWNCT VTCPURQTV RQTV 0WENGCT VTCPURQTV 4CP-DKPFKPI RTQVGKP øGUKEWNCT VTCPURQTV 4KDQUQOG CUUGODN[ /GVCDQNKUO 2hQURhCVG OGVCDQNKUO )62-DKPFKPI RTQVGKP
BRR1 BRR2 PRP16 DED1 CTK1, CTK2, CTK3 SRO7/SRO77
%GNN E[ENG CPF %GNN E[ENG &0# RTQEGUUKPI )62ð)&2-GZEhCPIG HCEVQT
CDC50
úGTð6hTð6[T 2TQVGKP MKPCUG
YRB2 DRS2 GTR1
LTE1 MCK1
4GNGXCPV TGHGTGPEGU
÷KP\GNGT GV CN. ÿþ999ý %TCKI CPF JCEQDUGP ÿþ985ý õGG CPF $CUGTIC ÿþ997ý 4KROCUVGT GV CN. ÿþ993ý 0QDNG CPF )WVhTKG ÿþ99öý úVGTPGT GV CN. ÿþ995ý õGhOCP GV CN. ÿþ999ý 0QIWEhK GV CN. ÿþ997; þ999ý 4KROCUVGT GV CN. ÿþ993ý 0CMCUhKOC GV CN. ÿþ99öý; FG JGUWU (GTTGKTC GV CN. ÿüûûþý 4CFLK GV CN. ÿüûûþý úhKTC[COC GV CN. ÿþ99öý 2KCQ GV CN. ÿþ999ý; FG JGUWU (GTTGKTC GV CN. ÿüûûþý
Industrial strains. # FKHHGTGPVKCN GZRTGUUKQP CPCN[UKU hCU DGGP ECTTKGF QWV YKVh DCMGT•U [GCUV WPFGT EQNF CPF HTGG\G UVTGUU EQPFKVKQPU, FGOQPUVTCVKPI VhCV DCMGT•U
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
8þ
[GCUVU UhQY C URGEKHKE RCVVGTP QH IGPG GZRTGUUKQP KP TGURQPUG VQ C UhKHV VQ NQY VGORGTCVWTGU ÿ4QFTKIWG\-øCTICU GV CN. üûûüý. #NVhQWIh IGPG TGRTGUUKQP YCU OQTG EQOOQP HQT HTGG\G-UhQEMGF EGNNU DGVYGGP ôû CPF öû E&0#, DCPFU YGTG KPFWEGF D[ DQVh EQNF- CPF HTGG\G-UVTGUU. 6hG URGEKHKEKV[ QH VhG TGURQPUG YCU EQPHKTOGF HQT TPI1, YhKEh GPEQFGU VTKQUG-RhQURhCVG KUQOGTCUG, ERG10, VhG IGPG HQT CEGVQCEGV[N EQGP\[OG-# VhKQNCUG, CPF IMH1, C RTQVGKP KPXQNXGF KP RTQVGKP VTCPURQTV. +PVGTGUVKPIN[, VhG GZRTGUUKQP QH TPI1 CPF ERG10 YCU KPFWEGF KP HTGG\G-UVTGUUGF UCORNGU QH C VQNGTCPV EQOOGTEKCN DCMGT•U [GCUV, DWV PQV KP C UGPUKVKXG UVTCKP. (WTVhGTOQTG, ERG10 QXGTGZRTGUUKQP GPhCPEGF HTGG\G VQNGTCPEG KP S. cerevisiae. ©CTNKGT UVWFKGU YKVh C [GCUV UVGTQN CWZQVTQRh TGRQTVGF C RQUKVKXG EQTTGNCVKQP DGVYGGP GTIQUVGTQN CPF VQNGTCPEG VQ hGCV CPF GVhCPQN ÿúYCP CPF ÷CVUQP þ998ý. 6hGUG FCVC KPFKECVG VhCV CP[ RTQFWEV QH VhG OGXCNQPCVG RCVhYC[ EQWNF RNC[ C TQNG KP HTGG\G TGUKUVCPEG KP [GCUV. /KETQCTTC[ CPCN[UKU QH HTGG\G-VQNGTCPV XGTUWU HTGG\G-UGPUKVKXG UVTCKPU FGTKXGF HTQO EQOOGTEKCN DCMGT•U [GCUV hCU CNUQ DGGP ECTTKGF QWV KP QTFGT VQ KFGPVKH[ IGPGU EQPHGTTKPI VhKU RhGPQV[RG ÿ6CPIhG GV CN. üûûüý. %QORCTKUQP QH IGPG RTQHKNG GZRTGUUKQP, WPFGT HGTOGPVKPI EQPFKVKQPU, UhQYGF VhCV þ5 IGPGU YGTG NQYGT GZRTGUUGF, YhKNG ö7 YGTG WR-TGIWNCVGF KP VhG TGUKUVCPV UVTCKP ÿ#6ü5ý. 1PN[ UKZ IGPGU, HTQO YhKEh HKXG GPEQFG OGODTCPG QT EGNN YCNN TGNCVGF RTQVGKPU, YGTG HWTVhGT UVWFKGF. *QYGXGT, PGKVhGT KPFKXKFWCN QXGTGZRTGUUKQP PQT FGNGVKQP TGUWNVGF KP UKIPKHKECPV GHHGEVU QP HTGG\G VQNGTCPEG. 3.3.3.4 Strategies and targets for improvement of cryoresistance +V KU ENGCT HTQO QWT NKOKVGF MPQYNGFIG QH HTGG\G VQNGTCPEG VhCV VhKU VTCKV KPXQNXGU C EQQTFKPCVGF UGV QH OGEhCPKUOU, CPF VhGTGHQTG, KV KU NKMGN[ VhCV QPN[ C EQQTFKPCVGF CEVKXCVKQP QH CNN QH VhGO TGUWNV KP C FTCUVKE KORTQXGOGPV. /GCPYhKNG KPXGUVKICVKQP TGXGCNU PGY RNC[GTU, TGIWNCVQTU, CPF KPVGTCEVKQPU COQPI VhGO. úQOG UVTCVGIKGU CPF VCTIGVU, hGTGCHVGT QWVNKPGF, EQWNF hGNR VQ FGXGNQR UVTCKPU YKVh DGVVGT RTQRGTVKGU. +V KU PQV KPVGPFGF CU CP GZhCWUVKXG NKUV, DWV TCVhGT C FGUETKRVKQP QH UGXGTCN KPVGTGUVKPI GZCORNGU. (TGG\G UVTGUU CRRGCTU VQ UhCTG C EQOOQP TGURQPUG, CV NGCUV KP RCTV, YKVh QVhGT UVTGUUGU, NKMG FGUKEECVKQP, QUOQVKE, CPF QZKFCVKXG. 6hGTGHQTG, VCTIGVU OGPVKQPGF KP QVhGT UGEVKQPU VQ KORTQXG UVTGUU TGUKUVCPEG ÿUGEV. 3.3.ü.5ý CTG GZRGEVGF VQ hGNR HTGG\G VQNGTCPEG. Membrane fluidity. 2QN[WPUCVWTCVGF HCVV[ CEKFU RNC[ CP KORQTVCPV TQNG KP RTQVGEVKQP QH VhG EGNN OGODTCPG ÿúCMCOQVQ CPF /WTCVC üûûüý, C OCLQT VCTIGV QH HTGG\GVhCY UVTGUU. #EEQTFKPIN[, VhG ETGCVKQP QH [GCUV EGNNU YKVh hKIh NGXGNU QH RQN[WPUCVWTCVGF CEKFU EQWNF DG C TGCNKUVKE CRRTQCEh VQ KORTQXG OGODTCPG HNWKFKV[, CPF VhWU, HTGG\G VQNGTCPEG. *QYGXGT, S. cerevisiae QPN[ FKURNC[U •9 HCVV[ CEKF FGUCVWTCUG CEVKXKV[, GPEQFGF D[ OLE1, YhGTGD[ þö:þ CPF þ8:þ HCVV[ CEKFU CTG HQTOGF. +VU GZRTGUUKQP KU TGIWNCVGF CV VhG NGXGN QH VTCPUETKRVKQP CPF O40# UVCDKNKV[ D[ FKHHGTGPV HCEVQTU ÿ/CTVKP GV CN. üûûüý, COQPI VhGO, NQY VGORGTCVWTG ÿ0CMCICYC GV CN. üûûüý. 6hKU QDUGTXCVKQP UWIIGUVU VhCV VhG TGIWNCVKQP QH FGUCVWTCVKQP QH HCVV[ CEKFU KP Saccharomyces RNC[U C TQNG KP VQNGTCPEG VQ EQNF UVTGUU. •þü CPF •ö HCVV[ CEKF FGUCVWTCUGU CTG YKFGN[ GZVGPFGF KP hKIhGT RNCPVU CPF E[CPQDCEVGTKC ÿõQU CPF /WTCVC
8ü
(. 4CPFG\-)KN GV CN.
þ998ý, UQOG QH YhKEh hCXG DGGP UWEEGUUHWNN[ GZRTGUUGF KP YKNF S. cerevisiae ÿ2G[QW-0FK GV CN. üûûû; &[GT GV CN. üûûþ; /CTVsPG\-4KXCU J/, RGTUQPCN EQOOWPKECVKQPý. 4GEGPVN[, YG hCXG ENQPGF CPF QXGTGZRTGUUGF VYQ •þü HCVV[ CEKF FGUCVWTCUGU HTQO UWPHNQYGT KP DCMGTU• [GCUV. 2TGNKOKPCT[ EhCTCEVGTKUCVKQP QH VhG HCVV[ CEKF EQORQUKVKQP QH YhQNG VTCPUHQTOCPV EGNNU TGXGCNGF VhG RTGUGPEG QH UKIPKHKECPV COQWPVU QH þö:ü CPF þ8:ü HCVV[ CEKFU VQIGVhGT YKVh C EQPEQOKVCPV KPETGCUG KP VhG TCVKQ QH WPUCVWTCVGF VQ UCVWTCVGF HCVV[ CEKFU. 6hKU EhCPIG KP VhG HCVV[ CEKF RTQHKNG KPETGCUGF ITQYVh TCVG CV NQY VGORGTCVWTGU CPF EGNN UWTXKXCN FWTKPI HTGG\G-UVTGUU ÿ4QFTKIWG\-øCTICU ú, /CTVsPG\-4KXCU J/, 4CPFG\-)KN (, WPRWDNKUhGF TGUWNVUý. Aquaporins. #SWCRQTKPU CTG EhCPPGN RTQVGKPU VhCV RNC[ CP KORQTVCPV TQNG KP hKIhGT RNCPVU CPF CPKOCNU HCEKNKVCVKPI TCRKF YCVGT KPHNWZðGHHNWZ HQT OCKPVGPCPEG QH VWTIQT CPF EGNNWNCT XQNWOG ÿ$QPhKXGTU GV CN. þ998; /G[TKCN GV CN. üûûþý. S. cerevisiae RQUUGUU VYQ IGPGU YKVh UVTQPI UKOKNCTKV[ YKVh CSWCRQTKPU, AQY1 CPF AQY2 ÿ*QhOCPP GV CN. üûûûý. 0GXGTVhGNGUU, VhGKT Rh[UKQNQIKECN HWPEVKQP KP YCVGT hQOGQUVCUKU CPF QUOQTGIWNCVKQP TGOCKPU WPEGTVCKP ÿ/G[TKCN GV CN. üûûþý. +PVGTGUVKPIN[, KV hCU DGGP TGEGPVN[ GXKFGPEGF VhCV QXGTGZRTGUUKQP QH AQY1 CPF AQY2 KP KPFWUVTKCN DCMGT•U [GCUV, KORTQXGU HTGG\G VQNGTCPEG ÿ6CPIhG GV CN. üûûüý. +P CFFKVKQP, VTCPUHQTOCPVU UhQYGF C hKIhGT UWTXKXCN TCVKQ FWTKPI HTQ\GP-FQWIh UVQTCIG CPF KVU KPKVKCN HGTOGPVCVKQP ECRCEKV[ YCU PQV CHHGEVGF D[ CSWCRQTKPU QXGTGZRTGUUKQP. 6hGUG QDUGTXCVKQPU EQWNF KPFKECVG VhCV C TCRKF QUOQVKECNN[ FTKXGP GHHNWZ QH YCVGT FWTKPI VhG HTGG\KPI RTQEGUU TGFWEGU KEG ET[UVCN HQTOCVKQP CPF TGUWNVKPI EGNN FCOCIG ÿ6CPIhG GV CN. üûûüý. Antifreezing proteins. #PQVhGT UVTCVGI[ VQ GPhCPEG ET[QVQNGTCPEG KP [GCUV EQWNF DG VhG GZRTGUUKQP QH CPVKHTGG\G RTQVGKPU. 6hG CPVKHTGG\G RTQVGKPU ÿ#(2Uý CPF IN[EQRTQVGKPU HTQO RQNCT HKUh CPF VhG VhGTOCN h[UVGTGUKU RTQVGKPU HTQO CTEVKE KPUGEVU KPhKDKV VhG ITQYVh QH NCTIG KEG ET[UVCNU ÿTGET[UVCNK\CVKQP KPhKDKVKQPý VhCV RTGXGPV EGNNWNCT FCOCIG ÿ$CTTGVV üûûþý. #(2U HTQO ÷KPVGT (NQWPFGT hCXG DGGP GZRTGUUGF KP [GCUV CU C EhKOGTKE RTQVGKP. óGCUV EGNNU hCTDQWTKPI VhKU EQPUVTWEVKQP UhQYGF C VYQHQNF KPETGCUG KP UWTXKXCN CHVGT HTGG\KPI ÿ/EòQYP CPF ÷CTTGP þ99þý, YhKEh FGOQPUVTCVGU VhCV C HWPEVKQPCN RTQVGKP EQWNF DG GZRTGUUGF KP VhKU QTICPKUO. +P QTFGT VQ TGCEh KPETGCUGF GZRTGUUKQP NGXGNU, VCPFGO TGRGCVU QH VhG #(2ö IGPG hCU DGGP GHHKEKGPVN[ GZRTGUUGF CPF VhG CPVKHTGG\G CEVKXKV[ hCU DGGP OGCUWTGF KP VhG RTQFWEVU UGETGVGF VQ VhG OGFKWO ÿ&TKGFQPMU GV CN. þ995ý. 6hWU, C HCOKN[ QH TGNCVGF KPVTCEGNNWNCT UMKP V[RG QH CPVKHTGG\G RQN[RGRVKFGU HTQO ÷KPVGT (NQWPFGT VhCV KU WUGF VQ OCMG EGNNU EQNF-TGUKUVCPV hCU DGGP RCVGPVGF ÿ*GY CPF )QPI üûûþý. +P QWT NCDQTCVQT[, YG hCXG TGEGPVN[ QXGTGZRTGUUGF VhG OCLQT ÿ)ú-5ý CPVKHTGG\G RQN[RGRVKFG KUQNCVGF HTQO ITWDD[ UEWNRKP KP KPFWUVTKCN DCMGT•U [GCUV ÿ%hCMTCDCTVV[ GV CN. þ988ý, Myoxocephalus aenaeus. 6TCPUHQTOCPVU UhQYGF C UKIPKHKECPV KORTQXGOGPV KP EGNN UWTXKXCN CPF NGCXGPKPI ECRCEKV[ CHVGT HTQ\GP-UVQTCIG QH NKSWKF FQWIh OQFGN U[UVGO QT DTGCF FQWIh ÿ2CPCFGTQ J, 4CPFG\-)KN (, 2TKGVQ J#, WPRWDNKUhGF TGUWNVUý. Oxidative defence. #RRTQCEhGU HQT KORTQXGF ET[QTGUKUVCPEG KP DCMGT•U [GCUV EQWNF KPENWFG VhG GPIKPGGTKPI QH QZKFCVKXG FGHGPEG OGEhCPKUOU. +P S. cerevisiae, VhG VTCPUETKRVKQP HCEVQT óCRþR ÿúEhPGNN GV CN. þ99ü; òWIG CPF JQPGU þ99ôý hCU DGGP KORNKECVGF KP VhG QZKFCVKXG UVTGUU TGURQPUG. óCRþR EQPVTQNU C NCTIG QZKFCVKXG
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
83
UVTGUU TGURQPUG TGIWNQP QH CV NGCUV 3ü RTQVGKPU ÿõGG GV CN. þ999ý, KPENWFKPI GSH1 ÿÄ-INWVCO[NE[UVGKPG U[PVhGVCUGý, TRX2 ÿVhKQTGFQZKPý, TRR1 ÿVhKQTGFQZKP TGFWEVCUGý, GLR1 ÿINWVCVhKQPG TGFWEVCUGý, CTT1 ÿE[VQUQNKE ECVCNCUGý, CPF VhG UWRGTQZKFG FKUOWVCUGU SOD1 CPF SOD2. 6hWU, QXGTGZRTGUUKQP QH YAP1 EQWNF RTQXKFG EQQTFKPCVGF GZRTGUUKQP QH QZKFCVKXG FGHGPEG IGPGU OCMKPI VhG GPVKTG 41úFGVQZKHKECVKQP U[UVGO YQTM GHHKEKGPVN[.
3.4 PQP-Saccharomyces DCMGT•U [GCUV: Torulaspora delbrueckii &QWIh NGCXGPKPI CDKNKV[ hCU DGGP TGRQTVGF HQT [GCUV QVhGT VhCP S. cerevisiae. óGCUV UWEh CU Issatchenkia orientalis, Pichia membranaefaciens, CPF Torulaspora delbrueckii YGTG VhG OQUV CDWPFCPV PQP-Saccharomyces URGEKGU RTGUGPV KP hQOGOCFG EQTP CPF T[G DTGCF FQWIh ÿ*ChP CPF òCYCK þ99û; #NOGKFC CPF 2CKU þ99öCý. $GVYGGP VhGO, VhG [GCUV Torulaspora delbrueckii hCU DGGP TGEQIPK\GF CU HTGG\GVhCY VQNGTCPV ÿ#NOGKFC CPF 2CKU þ99öDý CPF UQOG T. delbrueckii UVTCKPU CTG EWTTGPVN[ EQOOGTEKCNKUGF KP JCRCP HQT TGIWNCT ÿ5ù UWETQUGý HTQ\GP FQWIh CRRNKECVKQPU. 3.4.1 /QTRJQNQIKECN CPF IGPGVKE EJCTCEVGTKUVKEU 6hG [GCUV T. delbrueckii, YhKEh KU HTGSWGPVN[ HQWPF KP CNEQhQNKE DGXGTCIGU, HTWKV LWKEGU, CPF hKIh-UWICT-EQPVCKPKPI HQQFU ÿ©UVGXG-íCT\QUQ GV CN. üûûþý KU QHVGP EQPUKFGTGF C URQKNCIG [GCUV ÿúEhWNNGT GV CN. üûûþý. 2h[NQIGPGVKECNN[, T. delbrueckii KU ITQWRGF KP CP KPFGRGPFGPV VCZQP VQ VhCV QH S. cerevisiae HTQO YhKEh KV YCU UGRCTCVGF D[ VhG YC[ VhG CUEWU CTQUG ÿøCWIhCP-/CTVKP CPF /CTVKPK þ998ý. +V UhQYU UKZ EhTQOQUQOGU TGUQNXGF D[ RWNUG-HKGNF IGN GNGEVTQRhQTGUKU KP VhG TCPIG QH 8ûû VQ þöûû òD ÿ1FC CPF 6QPQOWTC þ995ý, C INQDWNCT VQ QXCN EGNN OQTRhQNQI[, CPF UOCNNGT UK\G VhCP S. cerevisiae EQOOGTEKCN DCMGT•U [GCUV. +P CFFKVKQP, OQUV QH VhG DCMKPI UVTCKPU QH Torulaspora FKURNC[ C XGIGVCVKXG ITQYVh CU hCRNQKF ÿòWTV\OCP þ998ý. &GURKVG QH VhGUG FKHHGTGPEGU, S. cerevisiae CPF T. delbrueckii CTG IGPGCNQIKECNN[ ENQUGN[ TGNCVGF, CU KV hCU DGGP RTGXKQWUN[ TGRQTVGF D[ UGSWGPEG EQORCTKUQPU ÿJCOGU GV CN. þ99ö; 1FC GV CN. þ997; $GNNQEh GV CN. üûûûý. 3.4.2 $CMKPI CRRNKECVKQPU #U KV YCU TGHGTTGF CDQXG, QPG KORQTVCPV VTCKV VhCV FGVGTOKPGU VhG HGTOGPVCVKXG CEVKXKV[ QH EQOOGTEKCN UVTCKPU QH S. cerevisiae KP NGCP CPF NQY-UWETQUG DTGCF FQWIh KU VhG NGXGN QH OCNVCUG CPF KPXGTVCUG CEVKXKV[. T. delbrueckii UVTCKPU UhQY EQPUKFGTCDNG XCTKCVKQP KP VhGKT CDKNKVKGU VQ HGTOGPV CPF VQ CUUKOKNCVG ECTDQP EQORQWPFU, CU ICNCEVQUG QT OCNVQUG ÿòWTV\OCP þ998ý. 6hWU, OCP[ T. delbrueckii UVTCKPU KUQNCVGF HTQO PCVWTG, hCXG DGGP FKUECTFGF HQT DCMKPI CRRNKECVKQPU DGECWUG VhGKT RQQT OCN-
8ô
(. 4CPFG\-)KN GV CN.
VCUG CEVKXKV[ ÿ1FC CPF 6QPQOWTC þ993ý. 6hG UOCNN UK\G QH Torulaspora EGNNU hCU CNUQ hCORGTGF KVU KPFWUVTKCN RTQFWEVKQP DGECWUG KV IKXGU C RCTVKEWNCT FKUCFXCPVCIG FWTKPI EGNNU HKNVTCVKQP, KPETGCUKPI VKOG CPF EQUV KP VhG RTQEGUU. %QPUGSWGPVN[, FKHHGTGPV GHHQTVU YGTG OCFG VQ QXGTEQOG VhKU RTQDNGO D[ QDVCKPKPI NCTIGT-UK\G FKRNQKF EGNNU ÿúCUCMK CPF 1UhKOC þ987ý. &GURKVG VhGUG FTCYDCEMU, VhG GZEGRVKQPCN CDKNKV[ QH Torulaspora EGNNU VQ EQRG YKVh UGXGTCN UVTGUUGU hCU CKOGF VhG EhCTCEVGTK\CVKQP QH PGY KUQNCVGU D[ VhGKT RQVGPVKCN WVKNK\CVKQP KP VhG DTGCF OCMKPI KPFWUVT[ ÿ#NOGKFC CPF 2CKU þ99öCý. õGCXGPKPI CEVKXKV[ QH T. delbrueckii +)%53üþ CPF +)%53ü3 WRQP HTGG\G-VhCY CRRGCTGF VQ DG WPCNVGTGF KP EQORCTKUQP YKVh VhG FGETGCUG QH HGTOGPVCVKXG ECRCEKV[ ÿôûùý KP S. cerevisiae DCMGT•U [GCUV ÿ#NOGKFC CPF 2CKU þ99öDý. 6hKU TGUWNV EQWNF DG GZRNCKPGF, CV NGCUV KP RCTV, D[ VhG UNQY TCVG QH VTGhCNQUG OQDKNKUCVKQP HQWPF KP VhGUG UVTCKPU CPF VhGKT CDKNKV[ VQ TGURQPF TCRKFN[ VQ QUOQVKE UVTGUU. +PFGGF, VhG RTGXKQWU TGRQTV QH #NOGKFC CPF 2CKU ÿþ99öDý UhQYGF VhCV T. delbrueckii UVTCKPU CTG PQV CHHGEVGF D[ HGTOGPVCVKQP DGHQTG HTGG\KPI. #U GZRGEVGF, GHHGEVU QH HTQ\GP UVQTCIG YGTG FGNGVGTKQWU HQT CNN VhG UVTCKP VGUVGF, DGKPI NGUU RTQPQWPEGF KP T. delbrueckii ÿ*GTPCPFG\õQRG\ GV CN. üûû3Cý. +P CFFKVKQP, T. delbrueckii UVTCKPU UhQYGF 0Cî VQZKEKV[ TGUKUVCPEG CPF C IQQF NGCXGPKPI CEVKXKV[ KP CNN VhG HQTOWNCVKQPU VGUVGF, NGCP, TGIWNCT, CPF UYGGV FQWIh ÿ4CPFG\-)KN GV CN. üûûüý. 6hWU, VhG CRRNKECVKQP QH VhGUG UVTCKPU KP DTGCF OCMKPI YQWNF hCXG CFFKVKQPCN CFXCPVCIGU, UKPEG C UKPING UVTCKP ECP DG WUGF HQT OQUV QT CNN DCMKPI CRRNKECVKQPU YKVh VhG hKIhGUV RGTHQTOCPEG. 3.4.3 T. delbrueckii CU C OQFGN U[UVGO ïPNKMG Saccharomyces, T. delbrueckii UVTCKPU GZhKDKV CP GZEGRVKQPCN TGUKUVCPEG VQ QUOQVKE CPF HTGG\G-VhCY UVTGUU, YhKEh OCFG QH VhKU [GCUV C RQVGPVKCN OQFGN VQ GZRNQTG CPF WPFGTUVCPF VhG OGEhCPKUOU WPFGTN[KPI UVTGUU TGUKUVCPEG KP DCMGT•U [GCUV. +P CFFKVKQP, VhG RQVGPVKCN DKQVGEhPQNQIKECN CRRNKECVKQPU QH Torulaspora UVTCKPU TGPFGT VhGO CP KPVGTGUVKPI hQUV VQ GZRTGUU HQTGKIP RTQVGKPU VhCV ECP KORTQXG VhG URGEKHKE KPFWUVTKCN RTQEGUUGU KP YhKEh VhG[ CTG KPXQNXGF. *QYGXGT, XGT[ NKVVNG KU MPQYP CDQWV VhG Rh[UKQNQI[ CPF IGPGVKEU QH VhKU [GCUV CPF VhG VQQNU HQT KVU OCPKRWNCVKQP hCU PQV DGGP FGXGNQRGF. 4GEGPVN[, YG hCXG TGRQTVGF VhG EQPUVTWEVKQP QH C IGPQOKE NKDTCT[ HTQO VhKU QTICPKUO CPF VhG KUQNCVKQP, ENQPKPI, CPF RTGNKOKPCT[ EhCTCEVGTKUCVKQP QH T. delbrueckii URA3 IGPG, TdURA3 ÿ*GTPCPFG\-õQRG\ GV CN. üûûüý. ïTC- UVTCKPU QH VhG T. delbrueckii +)%53ü3 QDVCKPGF GKVhGT D[ 5-(1#-CKFGF UGNGEVKQP QT VTCPUHQTOCVKQP YKVh C 2%4-DCUGF FKUTWRVKQP ECUUGVVG CTG CNUQ CXCKNCDNG ÿ*GTPCPFG\-õQRG\ GV CN. üûû3Dý. (WTVhGTOQTG, UGSWGPEKPI QH QVhGTU CWZQVTQRhKE OCTMGTU, UWEh CU TdLEU2, TdHIS3, CPF TdTRP1 KU KP RTQITGUU ÿ#NNGT-#TTCP\ ©, 4CPFG\-)KN (, 2TKGVQ J#, WPRWDNKUhGF TGUWNVUý. (KPCNN[, YG CTG UVCTVKPI VQ KFGPVKH[ IGPGU QH Torulaspora EQPHGTTKPI ET[QVQNGTCPEG KP S. cerevisiae DCMGT•U [GCUV, YhKEh RTQOKUG VQ DG CP KORQTVCPV VQQN VQ WPFGTUVCPF VhG OQNGEWNCT DCUKU QH VhKU RhGPQV[RG.
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
85
3.ý %QPENWUKQPU +P VhG RCUV HGY [GCTU, TGEQODKPCPV &0# VGEhPQNQI[ hCU NGF VQ VhG CRRCTKVKQP QH PGY DCMGT•U [GCUV UVTCKPU YKVh QRVKOK\GF QT PQXGN RTQRGTVKGU CPF KV KU GZRGEVGF VhCV VhKU VQQN YKNN RTQFWEG C hWIG URGEVTWO QH URGEKCNK\GF [GCUV QH hKIh CFFGF XCNWG KP C PGCT HWVWTG ÿ2TKGVQ GV CN. üûû3ý. 6hGKT KPVTQFWEVKQP KP VhG OCPWHCEVWTKPI OCTMGV YKNN RTQFWEG C FTCOCVKE EhCPIG KP HQTOWNCVKQP, KPITGFKGPVU, QT RTQEGUUKPI EQPFKVKQPU EWTTGPVN[ WUGF KP VhG DCMKPI RTCEVKEG CPF YKNN RTQXKFG PGY GPF-RTQFWEVU YKVh GPhCPEGF HNCXQWT, VGZVWTGU, QT GZVGPFGF UhGNH NKHG. $WV VhG OCLQT DKQVGEhPQNQIKECN EhCNNGPIG KP VhKU CTGC KU VhG UKIPKHKECPV KORTQXGOGPV QH HGTOGPVCVKXG VTCKVU, QUOQVQNGTCPEG CPF ET[QTGUKUVCPEG, QH DCMGT•U [GCUV. 6hGTG KU UVTQPI GXKFGPEG VhCV VhGUG EQOOGTEKCNN[ TGNGXCPV VTCKVU CTG WPFGT VhG EQPVTQN QH C ITGCV PWODGT QH IGPGU CPF EGNNWNCT HWPEVKQPU. 6hWU, INQDCN Rh[UKQNQIKECN TGUGCTEh YKVhKP VhG EQPVGZV QH KPFWUVTKCN TGNGXCPV EQPFKVKQPU KU VhG QDXKQWU VQQN VQ WPFGTUVCPFKPI hQY OWNVKRNG IGPGU CTG KPXQNXGF KP [GCUV VQ CEEQORNKUh OGVCDQNKE RTQEGUUGU FGVGTOKPKPI VhG RGTHQTOCPEG QH DCMGT•U [GCUV. 1PN[ VhG MPQYNGFIG, CV VhG OQNGEWNCT NGXGN, QH OGVCDQNKE PGVYQTMU ECP hGNR WU VQ EhQQUG VTCPUETKRVKQPCN TGIWNCVQTU CU VCTIGVU VQ EQPVTQN C URGEKHKE VTCKV. *QYGXGT, YG OWUV MGGR KP OKPF VhG TGEGPV YQTM FQPG D[ õGG CPF EQNNGCIWGU ÿüûûüý YhQ KNNWUVTCVGF EGNNWNCT PGVYQTMU QH VTCPUETKRVKQPCN TGIWNCVQTU VhCV EQPVTQN GZRTGUUKQP QH QVhGT VTCPUETKRVKQPCN HCEVQTU. òGGRKPI C TGCNKUVKE RGTURGEVKXG, KORTQXGOGPV QH VhGUG EQOOGTEKCN EhCTCEVGTKUVKEU YKNN DG C FKHHKEWNV VCUM. 6hGTGHQTG, PQP-Saccharomyces [GCUVU YKVh C hKIh UVTGUU TGUKUVCPEG CRRGCT CU C RTQOKUKPI CNVGTPCVKXG HTQO C VGEhPQNQIKECN RQKPV QH XKGY.
#EMPQYNGFIGOGPVU 6hG CURGEVU QH QWT TGUGCTEh FGUETKDGF KP VhKU EhCRVGT YGTG UWRRQTVGF D[ VhG %QOKUKxP +PVGTOKPKUVGTKCN FG %KGPEKC [ 6GEPQNQIsC RTQLGEVU ÿ#õ+98-û35ö; 2#%6+ %11þ999#ñþ73; #)õüûûû-û5üô; #)õüûûþ-þüû3ý.
üGHGTGPEGU #FCOU J, 2WUMCU-4Q\UC ú, úKONCT J, ÷KNMG %/ ÿþ99üý #FCRVCVKQP CPF OCLQT EhTQOQUQOCN EhCPIGU KP RQRWNCVKQPU QH Saccharomyces cerevisiae. %WTT )GPGV üü:þ3-þ9 #IWKNGTC #, òNGKP *õ ÿþ993ý %hTQOQUQOG CDGTTCVKQPU KP UKORNGT GWMCT[QVGU. +P: òKTUEh +4 ÿGF.ý 6hG %CWUGU CPF %QPUGSWGPEGU QH %hTQOQUQOCN #DGTTCVKQPU. %4% 2TGUU, $QEC 4CVQP, RR 5þ-9û #IWKNGTC J, 2TKGVQ J# ÿüûûþý 6hG Saccharomyces cerevisiae CNFQUG TGFWEVCUG KU KORNKGF KP VhG OGVCDQNKUO QH OGVh[NIN[QZCN KP TGURQPUG VQ UVTGUU EQPFKVKQPU. %WTT )GPGV 39:ü73ü83 #NDGTV[P J, *QhOCPP ú, 6hGXGNGKP J/, 2TKQT $# ÿþ99ôý GPD1, YhKEh GPEQFGU IN[EGTQN-3RhQURhCVG FGh[FTQIGPCUG, KU GUUGPVKCN HQT ITQYVh WPFGT QUOQVKE UVTGUU KP Saccharomy-
8ö
(. 4CPFG\-)KN GV CN.
ces cerevisiae, CPF KVU GZRTGUUKQP KU TGIWNCVGF D[ VhG hKIh-QUOQNCTKV[ IN[EGTQN TGURQPUG RCVhYC[. /QN %GNN $KQN þô:ôþ35-ôþôô #NGRW\ 2/, %WPPKPIhCO ò÷, ©UVTWEh ( ÿþ997ý )NWEQUG TGRTGUUKQP CHHGEVU KQP hQOGQUVCUKU KP [GCUV VhTQWIh TGIWNCVKQP QH VhG UVTGUU-CEVKXCVGF ENA1 IGPG. /QN /KETQDKQN üö:9þ-98 #NGRW\ 2/, JQXCPQXKE #, 4GKUGT ø, #OOGTGT ) ÿüûûþý úVTGUU-KPFWEGF OCR MKPCUG *QIþ KU RCTV QH VTCPUETKRVKQP CEVKXCVKQP EQORNGZGU. /QN %GNN 7:7ö7-777 #NOGKFC /J, 2CKU %ú ÿþ99öCý %hCTCEVGTK\CVKQP QH VhG [GCUV RQRWNCVKQP HTQO VTCFKVKQPCN EQTP CPF T[G DTGCF FQWIhU. õGVV #RRN /KETQDKQN ü3:þ5ô-þ58 #NOGKFC /J, 2CKU %ú ÿþ99öDý õGCXGPKPI CDKNKV[ CPF HTGG\G VQNGTCPEG QH [GCUVU KUQNCVGF HTQO VTCFKVKQPCN EQTP CPF T[G DTGCF FQWIhU. #RRN ©PXKTQP /KETQDKQN öü:ôôûþ-ôôûô #NQPUQ-/QPIG 4, 4GCN ©, ÷QLFC +, $GDGNOCP J2, /CIGT ÷*, úKFGTKWU / ÿüûûþý *[RGTQUOQVKE UVTGUU TGURQPUG CPF TGIWNCVKQP QH EGNN YCNN KPVGITKV[ KP Saccharomyces cerevisiae UhCTG EQOOQP HWPEVKQPCN CURGEVU. /QN /KETQDKQN ôþ:7þ7-73û #RUG /2, $NWOYCNF © ÿüûûüý ©PIKPGGTKPI UCNV VQNGTCPEG KP RNCPVU. %WTT 1RKP $KQVGEhPQN þ3:þôö-þ5û #VVHKGNF 2ø ÿþ997ý úVTGUU VQNGTCPEG: VhG MG[ VQ GHHGEVKXG UVTCKPU QH KPFWUVTKCN DCMGT•U [GCUV. 0CVWTG $KQVGEhPQN þ5:þ35þ-þ357 #VVHKGNF 2ø, òNGVUCU ú ÿüûûûý *[RGTQUOQVKE UVTGUU TGURQPUG D[ UVTCKPU QH DCMGTU' [GCUVU KP hKIh UWICT EQPEGPVTCVKQP OGFKWO. õGVV #RRN /KETQDKQN 3þ:3ü3-3ü7 $CTTGVV J ÿüûûþý 6hGTOCN h[UVGTGUKU RTQVGKPU. +PV J $KQEhGO %GNN $KQN 33:þû5-þþ7 $GNN /, %CRQPG 4, 2CUhVCP +, õGXKV\MK #, ©PIGNDGTI & ÿüûûþý +UQNCVKQP QH h[RGTCEVKXG OWVCPVU QH VhG /#2 MKPCUG R38ð*QIþ VhCV CTG KPFGRGPFGPV QH /#2òò CEVKXCVKQP. J $KQN %hGO ü7ö:ü535þ-ü5358 $GNNQEh %, ìWGTQN #, )CTEsC /&, $CTTKQ © ÿüûûûý 2h[NQIGP[ QH VhG IGPWU Kluyveromyces KPHGTTGF HTQO VhG OKVQEhQPFTKCN E[VQEhTQOG-E QZKFCUG ++ IGPG. +PV J ú[UV ©XQN /KETQDKQN 5û:ôû5-ôþö $GPsVG\ 6, %CUVTGLxP (, )CUGPV-4COsTG\ J/, %QFxP #% ÿþ99öý &GXGNQROGPV QH PGY UVTCKPU HQT VhG HQQF KPFWUVT[. $KQVGEhPQN 2TQI þü:þô9-þö3 $NQO J, &G /CVVQU /J, )TKXGNN õ# ÿüûûûý 4GFKTGEVKQP QH VhG TGURKTQ-HGTOGPVCVKXG HNWZ FKUVTKDWVKQP KP Saccharomyces cerevisiae D[ QXGTGZRTGUUKQP QH VhG VTCPUETKRVKQP HCEVQT *CRôR. #RRN ©PXKTQP /KETQDKQN öö:þ97û-þ973 $NQODGTI # ÿüûûûý /GVCDQNKE UWTRTKUGU KP Saccharomyces cerevisiae FWTKPI CFCRVCVKQP VQ UCNKPG EQPFKVKQPU: SWGUVKQPU, UQOG CPUYGTU CPF C OQFGN. (©/ú /KETQDKQN õGVV þ8ü:þ-8 $NQODGTI #, #FNGT õ ÿþ99üý 2h[UKQNQI[ QH QUOQVQNGTCPEG KP HWPIK. #FX /KETQD 2h[U 33:þô5-üþü $QPhKXGTU /, %CTDTG[ J/, )QWNF úJ, #ITG 2 ÿþ998ý #SWCRQTKPU KP Saccharomyces-IGPGVKE CPF HWPEVKQPCN FKUVKPEVKQPU DGVYGGP NCDQTCVQT[ CPF YKNF-V[RG UVTCKPU. J $KQN %hGO ü73:ü75ö5-ü757ü %CTNUQP / ÿþ999ý )NWEQUG TGRTGUUKQP KP [GCUV. %WTT 1RKP /KETQDKQN ü:üûü-üû7 %CWUVQP *%, 4GP $, òQh úú, *CTDKUQP %6, òCPKP ©, JGPPKPIU ©), õGG 6+, 6TWG *õ, õCPFGT ©ú, óQWPI 4# ÿüûûþý 4GOQFGNKPI QH [GCUV IGPQOG GZRTGUUKQP KP TGURQPUG VQ GPXKTQPOGPVCN EhCPIGU. /QN $KQN %GNN þü:3ü3-337 %hCMTCDCTVV[ #, *GY %õ, úhGCTU /, (NGVEhGT ) ÿþ988ý 2TKOCT[ UVTWEVWTGU QH VhG CNCPKPGTKEh CPVKHTGG\G RQN[RGRVKFGU HTQO ITWDD[ UEWNRKP, Myoxocephalus aenaeus. %CP J íQQN öö:ôû3-ôû8 %QFxP #%, $GPsVG\ 6 ÿþ995ý øCTKCDKNKV[ QH VhG Rh[UKQNQIKECN HGCVWTGU CPF QH VhG PWENGCT CPF OKVQEhQPFTKCN IGPQOGU QH DCMGT•U [GCUVU. ú[UVGO #RRN /KETQDKQN þ8:3ô3-35ü
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
87
%QFxP #%, )CUGPV-4COsTG\ J/, $GPsVG\ 6 ÿþ995ý (CEVQTU YhKEh CHHGEV VhG HTGSWGPE[ QH URQTWNCVKQP CPF VGVTCF HQTOCVKQP KP Saccharomyces cerevisiae DCMGT•U [GCUVU. #RRN ©PXKTQP /KETQDKQN öþ:ö3û-ö38 %QFxP #%, $GPsVG\ 6, òQThQNC / ÿþ997ý %hTQOQUQOCN TGQTICPK\CVKQP FWTKPI OGKQUKU QH Saccharomyces cerevisiae DCMGT'U [GCUVU. %WTT )GPGV 3ü:üô7-59 %QFxP #%, $GPsVG\ 6, òQThQNC / ÿþ998ý %hTQOQUQOCN RQN[OQTRhKUO CPF CFCRVCVKQP VQ URGEKHKE KPFWUVTKCN GPXKTQPOGPVU QH Saccharomyces UVTCKPU. #RRN /KETQDKQN $KQVGEhPQN ô9:þ5ô-ö3 %QFxP #%, 4KPExP #/, /QTGPQ-/CVGQU /#, &GNICFQ-JCTCPC J, 4G[ /, õKOxP %, 4QUCFQ +ø, %WDGTQ $, 2GwCVG ñ, %CUVTGLxP (, $GPsVG\ 6 ÿüûû3ý. 0GY Saccharomyces cerevisiae DCMGT•U [GCUV FKURNC[KPI GPhCPEGF TGUKUVCPEG VQ HTGG\KPI. J #ITKE (QQF %hGO 5þ:ô83-ô9þ %QTPKUh-$QYFGP #, *QHOG[T J* ÿþ995ý &GVGTOKPCVKQP QH EQPVTQN EQGHHKEKGPVU KP KPVCEV OGVCDQNKE U[UVGOU. $KQEhGO J ü98:3ö7-375 %TCKI ©#, JCEQDUGP ò ÿþ985ý /WVCVKQPU KP EQIPCVG IGPGU QH Saccharomyces cerevisiae hUR7û TGUWNV KP TGFWEGF ITQYVh TCVGU CV NQY VGORGTCVWTGU. /QN %GNN $KQN 5:35þ7-35üô %TGURQ Jõ, *CNN /0 ÿüûûüý ©NWEKFCVKPI 614 UKIPCNKPI CPF TCRCO[EKP CEVKQP: NGUUQPU HTQO Saccharomyces cerevisiae. /KETQDKQN /QN $KQN 4GX öö:579-59þ %TGURQ Jõ, &CKEhQ ò, ïUhKOCTW 6, *CNN /0 ÿüûûþý 6hG )#6# VTCPUETKRVKQP HCEVQTU GLN3 CPF GAT1 NKPM 614 VQ UCNV UVTGUU KP Saccharomyces cerevisiae. J $KQN %hGO ü7ö:3ôôôþ-3ôôôô FC %QUVC /ú, úCPVQU *, )CNKPUMK ©# ÿþ998ý #P QXGTXKGY QH VhG TQNG CPF FKXGTUKV[ QH EQORCVKDNG UQNWVGU KP DCEVGTKC CPF CTEhCGC. +P: #PVTCPKMKCP ) ÿGFý $KQVGEhPQNQI[ QH GZVTGOQRhKNGU. #FXCPEGU KP DKQEhGOKECN GPIKPGGTKPIðDKQVGEhPQNQI[, XQN öþ. úRTKPIGT, 0GY óQTM, RR þþ7-þ53 FG )TQQV ©, $GDGNOCP J2, /CIGT ÷*, 2NCPVC 4J ÿüûûûý øGT[ NQY COQWPVU QH INWEQUG ECWUG TGRTGUUKQP QH VhG UVTGUU-TGURQPUKXG IGPG HSP12 KP Saccharomyces cerevisiae. /KETQDKQNQI[ þôö:3ö7-375 FG JGUWU (GTTGKTC /%, $CQ ñ, õCK\o ø, *QhOCPP ú ÿüûûþý 6TCPURQUQP OWVCIGPGUKU TGXGCNU PQXGN NQEK CHHGEVKPI VQNGTCPEG VQ UCNV UVTGUU CPF ITQYVh CV NQY VGORGTCVWTG. %WTT )GPGV ôû:ü7-39 &GSWKP ú ÿüûûþý 6hG RQVGPVKCN QH IGPGVKE GPIKPGGTKPI HQT KORTQXKPI DTGYKPI, YKPG-OCMKPI CPF DCMKPI [GCUVU. #RRN /KETQDKQN $KQVGEhPQN 5ö:577-588 &KFGTKEh J#, 4CCOUFQPM õ/, òTWEMGDGTI #õ, $GTFGP J#, XCP &CO ò ÿüûûþý 2h[UKQNQIKECN RTQRGTVKGU QH Saccharomyces cerevisiae HTQO YhKEh hGZQMKPCUG ++ hCU DGGP FGNGVGF. #RRN ©PXKTQP /KETQDKQN ö7:þ587-þ593 &TKGFQPMU 4#, 6QUEhMC *ó, øCP #NOMGTM J÷, úEhCHHGTU +/, øGTDCMGN J/ ÿþ995ý ©ZRTGUUKQP CPF UGETGVKQP QH CPVKHTGG\G RGRVKFGU KP VhG [GCUV Saccharomyces cerevisiae. óGCUV þþ:8ô9-8öô &[GT J/, %hCRKVCN &%, %CT[ J÷, 2GRRGTOCP #$ ÿüûûþý %hKNNKPI-UGPUKVKXG, RQUVVTCPUETKRVKQPCN TGIWNCVKQP QH C RNCPV HCVV[ CEKF FGUCVWTCUG GZRTGUUGF KP [GCUV. $KQEhGO $KQRh[U 4GU %QOOWP ü8ü:þûþ9-þûü5 ©UVGXG-íCT\QUQ $, )QUVKPECT #, $QDGV 4, ïTWDWTW (, ìWGTQN # ÿüûûþý úGNGEVKQP CPF OQNGEWNCT EhCTCEVGTK\CVKQP QH YKPG [GCUVU KUQNCVGF HTQO VhG É©N 2GPGFGUÌ CTGC. (QQF /KETQDKQN þ7:553-5öü ©UVTWEh ( ÿüûûûý úVTGUU-EQPVTQNNGF VTCPUETKRVKQP HCEVQTU, UVTGUU-KPFWEGF IGPGU CPF UVTGUU VQNGTCPEG KP DWFFKPI [GCUV. (©/ú /KETQDKQN 4GX üô:ôö9-ô8ö
88
(. 4CPFG\-)KN GV CN.
©XCPU +* ÿþ99ûý. óGCUV UVTCKPU HQT DCMKPI: TGEGPV FGXGNQROGPVU. +P: úRGPEGT J(6, úRGPEGT &/ ÿGFUý óGCUV 6GEhPQNQI[. úRTKPIGT-øGTNCI, *GKFGNDGTI, RR þ3-5ô (GTTKIPQ 2, 2QUCU (, òQGRR &, úCKVQ *, úKNXGT 2# ÿþ998ý 4GIWNCVGF PWENGQðE[VQRNCUOKE GZEhCPIG QH HOG1 /#2ò TGSWKTGU VhG KORQTVKP DGVC hQOQNQIU NMD5 CPF XPO1. ©/$1 J þ7:5öûö-5öþô (NKMYGGTV /6, øCP > íCPFGP õ, JCPUUGP ÷/, úVGGPUOC *ó, øCP &KLMGP J2, 2TQPM J6 ÿþ99öý 2[TWXCVG FGECTDQZ[NCUG: CP KPFKURGPUCDNG GP\[OG HQT ITQYVh QH Saccharomyces cerevisiae QP INWEQUG.óGCUV þü:üô7-ü57 (NKMYGGTV /6, òW[RGT /, XCP /CTKU #J, òQVVGT 2, XCP &KLMGP J2, 2TQPM J6 ÿþ999ý úVGCF[UVCVG CPF VTCPUKGPV-UVCVG CPCN[UKU QH ITQYVh CPF OGVCDQNKVG RTQFWEVKQP KP C Saccharomyces cerevisiae UVTCKP YKVh TGFWEGF R[TWXCVG-FGECTDQZ[NCUG CEVKXKV[. $KQVGEhPQN $KQGPI öö:ôü-5û )CPEGFQ J/ ÿþ998ý óGCUV ECTDQP ECVCDQNKVG TGRTGUUKQP. /KETQDKQN /QN $KQN 4GX öü:33ô3öþ )CUEh #2, úRGNNOCP 26, òCQ %/, %CTOGN-*CTGN 1, ©KUGP /$, úVQT\ ), $QVUVGKP &, $TQYP 21 ÿüûûûý )GPQOKE GZRTGUUKQP RTQITCOU KP VhG TGURQPUG QH [GCUV EGNNU VQ GPXKTQPOGPVCN EhCPIGU. /QN $KQN %GNN þþ:ôüôþ-ôü57 )CUGPV-4COsTG\ J/, %CUVTGLxP (, ìWGTQN #, 4COxP &, $GPsVG\ 6 ÿþ999ý )GPQOKE UVCDKNKV[ QH Saccharomyces cerevisiae DCMGT•U [GCUVU. ú[UVGO #RRN /KETQDKQN üü:3ü9-3ôû )CZKQNC 4, FG õCTTKPQC +(, øKNNCNDC J/, úGTTCPQ 4 ÿþ99üý # PQXGN CPF EQPUGTXGF UCNVKPFWEGF RTQVGKP KU CP KORQTVCPV FGVGTOKPCPV QH UCNV VQNGTCPEG KP [GCUV. ©/$1 J þþ:3þ57-3þöô )oNKPCU 2, (KUGV ), ÷KNNGOQV %, )QWNGV J ÿþ99þý õKRKF EQPVGPV CPF ET[QVQNGTCPEG QH DCMGT•U [GCUV KP HTQ\GP FQWIhU. #RRN ©PXKTQP /KETQDKQN 57:ôö3-ôö8 )oNKPCU 2, õCIKOQPKpTG /, &WDQTF % ÿþ993ý $CMGT'U [GCUV UCORNKPI CPF HTQ\GP FQWIh UVCDKNKV[. %GTGCN %hGO 7û:üþ9-üü5 )QPnCNXGU 2, 2NCPVC 4J ÿþ998ý úVCTVKPI WR [GCUV IN[EQN[UKU. 6TGPFU /KETQDKQN ö:3þô-3þ9 ){TPGT ÷, &WTEhUEhNCI ©, ÷QNH J, $TQYP ©õ, #OOGTGT ), 4WKU *, úEhWNNGT % ÿüûûüý #EWVG INWEQUG UVCTXCVKQP CEVKXCVGU VhG PWENGCT NQECNK\CVKQP UKIPCN QH C UVTGUU-URGEKHKE [GCUV VTCPUETKRVKQP HCEVQT. ©/$1 J üþ:þ35-þôô )WUVKP /%, #NDGTV[P J, #NGZCPFGT /, &CXGPRQTV ò ÿþ998ý /#2 MKPCUG RCVhYC[U KP VhG [GCUV Saccharomyces cerevisiae. /KETQDKQN /QN $KQN 4GX öü:þüöô-þ3ûû *ChP ó-ú, òCYCK * ÿþ99ûý +UQNCVKQP CPF EhCTCEVGTK\CVKQP QH HTGG\G-VQNGTCPV [GCUVU HTQO PCVWTG CXCKNCDNG HQT VhG HTQ\GP-FQWIh OGVhQF. #ITKE $KQN %hGO 5ô:8ü9-83þ *CORUG[ / ÿþ997ý # TGXKGY QH RhGPQV[RGU KP Saccharomyces cerevisiae. óGCUV þ3:þû99þþ33 *CTFKG &), %CTNKPI &, %CTNUQP / ÿþ998ý 6hG #/2-CEVKXCVGFðú0(þ RTQVGKP MKPCUG UWDHCOKN[: OGVCDQNKE UGPUQTU QH VhG GWMCT[QVKE EGNN! #PPW 4GX $KQEhGO ö7:8üþ-855 *CTQ 4, )CTEKCFGDNCU $, 4QFTKIWG\-0CXCTTQ # ÿþ99þý # PQXGN 2-V[RG #62CUG HTQO [GCUV KPXQNXGF KP UQFKWO VTCPURQTV. (©$ú õGVV ü9þ:þ89-þ9þ *GTOGU-õKOC /, úVQTG[ ò$ ÿþ993ý #PVKQZKFCPV FGHGPEGU KP VhG VQNGTCPEG QH HTGG\KPI CPF CPQZKC D[ ICTVGT UPCMGU. #O J 2h[UKQN üö5:4öôö-4ö5ü *GTPCPFG\-õQRG\ /J, 2TKGVQ J#, 4CPFG\-)KN ( ÿüûûüý +UQNCVKQP CPF EhCTCEVGTK\CVKQP QH VhG IGPG URA3 GPEQFKPI VhG QTQVKFKPG-5ê-RhQURhCVG FGECTDQZ[NCUG HTQO Torulaspora delbrueckii. óGCUV þ9:þô3þ-þô35 *GTPCPFG\-õQRG\ /J, 2TKGVQ J#, 4CPFG\-)KN ( ÿüûû3Cý 1UOQVQNGTCPEG CPF NGCXGPKPI CDKNKV[ KP UYGGV CPF HTQ\GP UYGGV FQWIh. %QORCTCVKXG CPCN[UKU DGVYGGP Torulaspora del-
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
89
brueckii CPF Saccharomyces cerevisiae DCMGT•U [GCUV UVTCKPU. #PVQPKG XCP õGGWYGPhQGM ÿKP RTGUUý *GTPCPFG\-õQRG\ /J, 2TKGVQ J#, 4CPFG\-)KN ( ÿüûû3Dý ïTC- hQUV UVTCKPU HQT IGPGVKE OCPKRWNCVKQP CPF hGVGTQNQIQWU GZRTGUUKQP QH Torulaspora delbrueckii. +PV J (QQF /KETQDKQN ÿKP RTGUUý *GY %, )QPI í ÿüûûþý +PVTCEGNNWNCT CPVKHTGG\G RQN[RGRVKFGU CPF PWENGKE CEKFU. ïPKVGF úVCVGU 2CVGPV ïú ö,3û7,ûüû $þ *KIIKPU ø, $TCKFYQQF /, $GNN 2, $KUUKPIGT 2, &CYGU +÷, #VVHKGNF 2ø ÿþ999ý )GPGVKE GXKFGPEG VhCV hKIh PQPKPFWEGF OCNVCUG CPF OCNVQUG RGTOGCUG CEVKXKVKGU, IQXGTPGF D[ MALx3-GPEQFGF VTCPUETKRVKQPCN TGIWNCVQTU, FGVGTOKPG GHHKEKGPE[ QH ICU RTQFWEVKQP D[ DCMGT•U [GCUV KP WPUWICTGF FQWIh. #RRN ©PXKTQP /KETQDKQN ö5:ö8û-ö85 *KIIKPU øJ, $GNN 2Jõ, &CYGU +÷, #VVHKGNF 2ø ÿüûûþý )GPGTCVKQP QH C PQXGN Saccharomyces cerevisiae UVTCKP VhCV GZhKDKVU UVTQPI OCNVQUG WVKNK\CVKQP CPF h[RGTQUOQVKE TGUKUVCPEG WUKPI PQPTGEQODKPCPV VGEhPKSWGU. #RRN ©PXKTQP /KETQDKQN ö7:ô3ôö-ô3ô8 *KPPGDWUEh #), 0CVCTCLCP ò ÿüûûüý )EPôR, C OCUVGT TGIWNCVQT QH IGPG GZRTGUUKQP, KU EQPVTQNNGF CV OWNVKRNG NGXGNU D[ FKXGTUG UKIPCNU QH UVCTXCVKQP CPF UVTGUU. ©WMCT[QV %GNN þ:üü-3ü *KTCUCYC 4, óQMQKICYC ò ÿüûûþý õGCXGPKPI CDKNKV[ QH DCMGT'U [GCUV GZRQUGF VQ h[RGTQUOQVKE OGFKC. (©/ú /KETQDKQN õGVV þ9ô:þ59-þöü *KTCUCYC 4, óQMQKICYC ò, +UQDG ó, òCYCK * ÿüûûþý +ORTQXKPI VhG HTGG\G VQNGTCPEG QH DCMGT•U [GCUV D[ NQCFKPI YKVh VTGhCNQUG. $KQUEK $KQVGEhPQN $KQEhGO ö5:5üü-5üö *QhOCPP ú ÿüûûüý 1UOQVKE UVTGUU UKIPCNNKPI CPF QUOQCFCRVCVKQP KP [GCUVU. /KETQDKQN /QN $KQN 4GX öö:3ûû-37ü *QhOCPP ú, $KNN 4/, òC[KPIQ ), 2TKQT $# ÿüûûûý /KETQDKCN /+2 EhCPPGNU. 6TGPFU /KETQDKQN 8:33-38 +OCK 4, %hCPI õ, 1hVC #, $TC[ ©#, 6CMCIK / ÿþ99öý # NGC-ENCUU IGPG QH VQOCVQ EQPHGTU UCNV CPF HTGG\KPI VQNGTCPEG YhGP GZRTGUUGF KP Saccharomyces cerevisiae. )GPG þ7û:üô3-üô8 +PQWG ó, úCRKTUVGKP *&, $WUhWM ÷ ÿþ995ý úVWFKGU QP HTQ\GP FQWIhU. +ø. ©HHGEV QH UhQTVGPKPI U[UVGOU QP DCMKPI CPF ThGQNQIKECN RTQRGTVKGU. %GTGCN %hGO 7ü:üüþ-üüö +PQW[G / ÿþ999ý %QNF-UhQEM TGURQPUG CPF CFCRVCVKQP. J /QN /KETQDKQN $KQVGEhPQN þ:þ9þ JCOGU ú#, %QNNKPU /&, 4QDGTVU +0 ÿþ99öý ïUG QH CP T40# KPVGTPCN VTCPUETKDGF URCEGT TGIKQP VQ FKUVKPIWKUh Rh[NQIGPGVKECNN[ TGNCVGF URGEKGU QH VhG IGPGTC Zygosaccharomyces CPF Torulaspora. +PV J ú[UV $CEVGTKQN ôö:þ89-þ9ô. JQhPUVQP / ÿþ999ý (GCUVKPI, HCUVKPI CPF HGTOGPVKPI. )NWEQUG UGPUKPI KP [GCUV CPF QVhGT EGNNU. 6TGPFU )GPGV þ5:ü9-33 òCWN ú%, 1DWEhK ò, +YChCUhK *, òQOCVUW ó ÿþ99üý %T[QRTQVGEVKQP RTQXKFGF D[ hGCV UhQEM VTGCVOGPV KP Saccharomyces cerevisiae. %GNN /QN $KQN 38:þ35-þô3 òKO J, #NK\CFGh 2, *CTFKPI 6, *GHPGT-)TCXKPM #, òNKQPUM[ &J ÿþ99öý &KUTWRVKQP QH VhG [GCUV ATH1 IGPG EQPHGTU DGVVGT UWTXKXCN CHVGT FGh[FTCVKQP, HTGG\KPI CPF GVhCPQN UhQEM: RQVGPVKCN EQOOGTEKCN CRRNKECVKQPU. #RRN ©PXKTQP /KETQDKQN öü:þ5ö3-þ5ö9 òNGKP %J, 4CUOWUUGP JJ, 4QPPQY $, 1NUUQP õ, 0KGNUGP J ÿþ999ý +PXGUVKICVKQP QH VhG KORCEV QH MIG1 CPF MIG2 QP VhG Rh[UKQNQI[ QH Saccharomyces cerevisiae. J $KQVGEhPQN ö8:þ97-üþü òQ %*, )CDGT 4( ÿþ99þý TRK1 CPF TRK2 GPEQFGU UVTWEVWTCNN[ TGNCVGF òî VTCPURQTVGTU KP Saccharomyces cerevisiae. /QN %GNN $KQN þþ:ôüöö-ôü73 òQPFQ ò, +PQW[G / ÿþ99þý TIP1, C EQNF-UhQEM KPFWEKDNG IGPG QH Saccharomyces cerevisiae. J $KQN %hGO üöö:þ7537-þ75ôô
9û
(. 4CPFG\-)KN GV CN.
òQYCNUMK õ4í, òQPFQ ò, +PQW[G / ÿþ995ý %QNF-UhQEM KPFWEVKQP QH C HCOKN[ QH TIP1TGNCVGF RTQVGKPU CUUQEKCVGF YKVh VhG OGODTCPG KP Saccharomyces cerevisiae. /QN /KETQDKQN þ5:3ôþ-353 òWIG ú, JQPGU 0 ÿþ99ôý YAP1 FGRGPFGPV CEVKXCVKQP QH TRX2 KU GUUGPVKCN HQT VhG TGURQPUG QH Saccharomyces cerevisiae VQ QZKFCVKXG UVTGUU D[ h[FTQRGTQZKFGU. ©/$1 J þ3:ö55-ööô òWTV\OCP %2 ÿþ998ý Torulaspora õKPFPGT. +P òWTV\OCP %2, (GNN J÷ ÿGFUý 6hG óGCUVU. # VCZQPQO[E UVWF[. ©NUGXKGT úEKGPEG $.ø., #OUVGTFCO, RR ôûô-ôû8 õCTUUQP ò, ©TKMUUQP 2, #PUGNN 4, #FNGT õ ÿþ993ý # IGPG GPEQFKPI sn-IN[EGTQN 3- RhQURhCVG FGh[FTQIGPCUG ÿ0#&îý EQORNGOGPVU CP QUOQUGPUKVKXG OWVCPV QH Saccharomyces cerevisiae. /QN /KETQDKQN þû:þþûþ-þþþþ õCVVGTKEh /, ÷CVUQP /& ÿþ993ý ©XKFGPEG HQT C FWCN QUOQTGIWNCVQT[ OGEhCPKUO KP VhG [GCUV Saccharomyces cerevisiae. $KQEhGO $KQRh[U 4GU %QOOWP þ9þ:þþþþ-þþþ7 õGG $0, ©NKQP ©# ÿþ999ý 6hG /#2òòò úVGþþ TGIWNCVGU XGIGVCVKXG ITQYVh VhTQWIh C MKPCUG ECUECFG QH UhCTGF UKIPCNKPI EQORQPGPVU. 2TQE 0CVN #ECF úEK ïú# 9ö:þüö79þüö8ô õGG J, )QFQP %, õCIPKGN ), úRGEVQT &, )CTKP J, õCDCTTG J, 6QNGFCPQ /$ ÿþ999ý óCRþ CPF úMP7 EQPVTQN VYQ URGEKCNK\GF QZKFCVKXG UVTGUU TGURQPUG TGIWNQPU KP [GCUV. J $KQN %hGO ü7ô:þöûôû-þöûôö õGG úJ, $CUGTIC úJ ÿþ997ý (WPEVKQPCN UGRCTCVKQP QH RTG-T40# RTQEGUUKPI UVGRU TGXGCNGF D[ VTWPECVKQP QH VhG ï3 UOCNN PWENGQNCT TKDQPWENGQRTQVGKP EQORQPGPV, /RRþû. 2TQE 0CVN #ECF úEK ïú# 9ô:þ353ö-þ35ôþ õGG 6+, 4KPCNFK 0J, 4QDGTV (, 1FQO &6, $CT-JQUGRh í, )GTDGT )ò, *CPPGVV 0/, *CTDKUQP %6, 6hQORUQP %/, úKOQP +, íGKVNKPIGT J, JGPPKPIU ©), /WTTC[ *õ, )QTFQP &$, 4GP $, ÷[TKEh JJ, 6CIPG J-$, øQNMGTV 6õ, (TCGPMGN ©, )KHHQTF &ò, óQWPI 4# ÿüûûüý 6TCPUETKRVKQPCN TGIWNCVQT[ PGVYQTMU KP Saccharomyces cerevisiae. úEKGPEG ü98:7998ûô õGhOCP ò, 4QUUK ), #FCOQ J©, $TGPPYCNF 2 ÿþ999ý óGCUV hQOQNQIWGU QH VQOQU[P CPF lethal giant larvae HWPEVKQP KP GZQE[VQUKU CPF CTG CUUQEKCVGF YKVh VhG RNCUOC OGODTCPG ú0#4©, úGE9. J %GNN $KQN þôö:þü5-þôû õGYKU J), õGCTOQPVh 42, ÷CVUQP ò ÿþ993ý 4QNG QH ITQYVh RhCUG CPF GVhCPQN KP HTGG\GVhCY TGUKUVCPEG QH Saccharomyces cerevisiae. #RRN ©PXKTQP /KETQDKQN 59:þûö5-þû7þ õGYKU J), õGCTOQPVh 42, #VVHKGNF 2ø, ÷CVUQP ò ÿþ997ý úVTGUU EQ-VQNGTCPEG CPF VTGhCNQUG EQPVGPV KP DCMKPI UVTCKPU QH Saccharomyces cerevisiae. J +PF /KETQDKQN $KQVGEhPQN þ8:3û-3ö õQU &#, /WTCVC 0 ÿþ998ý úVTWEVWTG CPF GZRTGUUKQP QH HCVV[ CEKF FGUCVWTCUGU. $KQEhKO $KQRh[U #EVC þ39ô:3-þ5 õQWKU ©J, 0CWOQXC ©ú, õGG #, 0CWOQX ), *CDGT J© ÿþ99ôý 6hG EhTQOQUQOG GPF KP [GCUV: KVU OQUCKE PCVWTG CPF KPHNWGPEG QP TGEQODKPCVKQPCN F[PCOKEU. )GPGVKEU þ3ö:789-8ûü õW[VGP ò, #NDGTV[P J, úMKDDG ÷(, 2TKQT $#, 4COQU J, 6hGXGNGKP J/, *QhOCPP ú ÿþ995ý (RUþ, C [GCUV OGODGT QH VhG /+2 HCOKN[ QH EhCPPGN RTQVGKPU, KU C HCEKNKVCVQT QH IN[EGTQN WRVCMG CPF GHHNWZ CPF KU KPCEVKXG WPFGT QUOQVKE UVTGUU. ©/$1 J þô:þ3öû-þ37þ /CGFC 6, ÷WTINGT-/WTRh[ ú/, úCKVQ * ÿþ99ôý # VYQ-EQORQPGPV U[UVGO VhCV TGIWNCVGU CP QUOQUGPUKPI /#2 MKPCUG ECUECFG KP [GCUV. 0CVWTG þ9:üôü-üô5 /CGFC 6, 6CMGMCYC /, úCKVQ * ÿþ995ý #EVKXCVKQP QH [GCUV 2$úü /#2òò D[ /2òòòU QT D[ DKPFKPI QH CP ú*3-EQPVCKPKPI QUOQUGPUQT. úEKGPEG üö9:55ô-558 /CIGT ÷*, 2NCPVC 4J ÿþ99þý %QQTFKPCVG GZRTGUUKQP QH TKDQUQOCN RTQVGKP IGPGU KP [GCUV CU C HWPEVKQP QH EGNNWNCT ITQYVh TCVG. /QN %GNN $KQEhGO þûô:þ8þ-þ87 /CIGT ÷*, (GTTGKTC / ÿþ993ý úVTGUU TGURQPUG QH [GCUV. $KQEhGO J ü9û:þ-þ3
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
9þ
/áTSWG\ J#, 2CUEWCN-#hWKT #, 2TQHV /, úGTTCPQ 4 ÿþ998ý 6hG úUPö-6WRþ TGRTGUUQT EQORNGZ QH Saccharomyces cerevisiae KU KPXQNXGF KP VhG QUOQVKE KPFWEVKQP QH *1)FGRGPFGPV CPF KPFGRGPFGPV IGPGU. ©/$1 J þ7:ü5ô3-ü553 /CTVKP %©, 1h %-ú, òCPFCUCO[ 2, %hGNNCRC 4, øGOWNC / ÿüûûüý óGCUV FGUCVWTCUGU. $KQEhGO úQE 6TCPU 3û:þû8û-þû8ü /CVhGQU &2, òKPIUDWT[ 6J, #hUCP ïú, %WPPKPIhCO ò÷ ÿþ997ý 6EPþRð%T\þR, C ECNEKPGWTKP-FGRGPFGPV VTCPUETKRVKQP HCEVQT VhCV FKHHGTGPVKCNN[ TGIWNCVGU IGPG GZRTGUUKQP KP Saccharomyces cerevisiae. )GPGU &GX þþ:3ôô5-3ô58 /E%CTVPG[ 44, úEhOKFV /% ÿüûûþý 4GIWNCVKQP QH úPHþ MKPCUG. #EVKXCVKQP TGSWKTGU RhQURhQT[NCVKQP QH VhTGQPKPG üþû D[ CP WRUVTGCO MKPCUG CU YGNN CU C FKUVKPEV UVGR OGFKCVGF D[ VhG úPHô UWDWPKV. J $KQN %hGO ü7ö:3öôöû-3öôöö /EòQYP 4õ, ÷CTTGP )J ÿþ99þý ©PhCPEGF UWTXKXCN QH [GCUV GZRTGUUKPI CP CPVKHTGG\G IGPG CPCNQIWG CHVGT HTGG\KPI. %T[QDKQNQI[ ü8:ô7ü-ô8ü /G[TKCN ø, õCK\o ø, )QDKP 4, 4KRQEhG 2, *QhOCPP ú, 6CEPGV ( ÿüûûþý ©ZKUVGPEG QH C VKIhVN[ TGIWNCVGF YCVGT EhCPPGN KP Saccharomyces cerevisiae. ©WT J $KQEhGO üö8:33ô3ô3 /QTCFCU-(GTTGKTC 2, %QUVC ø ÿüûûûý #FCRVKXG TGURQPUG QH VhG [GCUV Saccharomyces cerevisiae VQ TGCEVKXG QZ[IGP URGEKGU: FGHGPEGU, FCOCIG CPF FGCVh. 4GFQZ 4GRQTV 5:ü77ü85 /QTGPQ (, *GTTGTQ 2 ÿüûûüý 6hG hGZQMKPCUG ü-FGRGPFGPV INWEQUG UKIPCN VTCPUFWEVKQP RCVhYC[ QH Saccharomyces cerevisiae. (©/ú /KETQDKQN 4GX üö:83-9û /QTKVC ó, 0CMCOQTK ú, 6CMCIK * ÿüûû3ý õ-2TQNKPG CEEWOWNCVKQP CPF HTGG\G VQNGTCPEG KP Saccharomyces cerevisiae CTG ECWUGF D[ C OWVCVKQP KP VhG PRO1 IGPG GPEQFKPI ÄINWVCO[N MKPCUG. #RRN ©PXKTQP /KETQDKQN ö9:üþü-üþ9 /QTTKU )J, %QWNUQP )©, %NCTMG òJ ÿþ988ý (TGG\KPI KPLWT[ KP Saccharomyces cerevisiae. 6hG GHHGEVU QH ITQYVh EQPFKVKQPU. %T[QDKQNQI[ ü5:ô7þ-ô7ü /WTCMCOK ó, óQMQKICYC ò, òCYCK (, òCYCK * ÿþ99öý õKRKF EQORQUKVKQP QH EQOOGTEKCN DCMGT•U [GCUVU hCXKPI FKHHGTGPV HTGG\G-VQNGTCPEG KP HTQ\GP FQWIh. $KQUEK $KQVGEh $KQEhGO öû:þ87ô-þ87ö /[GTU &ò, #VVHKGNF 2ø ÿþ999ý +PVTCEGNNWNCT EQPEGPVTCVKQP QH GZQIGPQWU IN[EGTQN KP Saccharomyces cerevisiae RTQXKFGU HQT KORTQXGF NGCXGPKPI QH HTQ\GP UYGGV FQWIhU. (QQF /KETQDKQN þö:ô5-5þ /[GTU &ò, õCYNQT &6/, #VVHKGNF 2ø ÿþ997ý +PHNWGPEG QH KPXGTVCUG CEVKXKV[ CPF IN[EGTQN U[PVhGUKU CPF TGVGPVKQP QP HGTOGPVCVKQP QH OGFKC YKVh hKIh UWICT EQPEGPVTCVKQP D[ Saccharomyces cerevisiae. #RRN ©PXKTQP /KETQDKQN ö3:þô5-þ5û /[GTU &ò, JQUGRh ø/, 2GhO ú, )CNXCIPQ /, #VVHKGNF 2ø ÿþ998ý õQCFKPI QH Saccharomyces cerevisiae YKVh IN[EGTQN NGCFU VQ GPhCPEGF HGTOGPVCVKQP KP UYGGV DTGCF FQWIhU. (QQF /KETQDKQN þ5:5þ-58 0CMCICYC ú, 1WEhK ò ÿþ99ôý %QPUVTWEVKQP HTQO C UKPING RCTGPV QH DCMGT•U [GCUV UVTCKPU YKVh hKIh HTGG\G VQNGTCPEG CPF HGTOGPVCVKXG CEVKXKV[ KP DQVh NGCP CPF UYGGV FQWIhU. #RRN ©PXKTQP /KETQDKQN öû:3ô99-35ûü 0CMCICYC ó, úCMWOQVQ 0, òCPGMQ ó, *CTCUhKOC ú ÿüûûüý /ICüR KU C RWVCVKXG UGPUQT HQT NQY VGORGTCVWTG CPF QZ[IGP VQ KPFWEG OLE1 VTCPUETKRVKQP KP Saccharomyces cerevisiae. $KQEhGO $KQRh[U 4GU %QOOWP ü9þ:7û7-7þ3 0CMCUhKOC 0, *C[CUhK 0, 0QIWEhK ©, 0KUhKOQVQ 6 ÿþ99öý 2WVCVKXG )62CUG )VTþR IGPGVKECNN[ KPVGTCEVU YKVh VhG 4CP)62CUG E[ENG KP Saccharomyces cerevisiae. J %GNN úEK þû9:ü3þþ-ü3þ8
9ü
(. 4CPFG\-)KN GV CN.
0CWOQX )+, 0CWOQXC ©ú, õCPVVQ 4#, õQWKU ©J, òQThQNC / ÿþ99üý )GPGVKE hQOQNQI[ DGVYGGP Saccharomyces cerevisiae CPF KVU UKDNKPI URGEKGU S. paradoxus CPF S. bayanus: GNGEVTQRhQTGVKE MCT[QV[RGU. óGCUV 8:599-öþü 0CXCU /#, %GTFCP ú, )CPEGFQ J/ ÿþ993ý (WVKNG E[ENGU KP S. cerevisiae UVTCKPU GZRTGUUKPI INWEQPGQIGPKE GP\[OGU FWTKPI ITQYVh QP INWEQUG. 2TQE 0CVN #ECF úEK ïú# 9û:þü9ûþü9ô 0QDNG ú/, )WVhTKG % ÿþ99öý +FGPVKHKECVKQP QH PQXGN IGPGU TGSWKTGF HQT [GCUV RTG-O40# URNKEKPI D[ OGCPU QH EQNF-UGPUKVKXG OWVCVKQPU. )GPGVKEU þô3:ö7-8û 0QIWEhK ©, òC[CUhK 0, 0CMCUhKOC 0, 0KUhKOQVQ 6 ÿþ997ý óTDüR, C 0WRüR-TGNCVGF [GCUV RTQVGKP, hCU C HWPEVKQPCN QXGTNCR YKVh 4PCþR, C [GCUV 4CP-)62CUG-CEVKXCVKPI RTQVGKP. /QN %GNN $KQN þ7:üü35-üüôö 0QIWEhK ©, úC[KVQ ó, úCDGT ú, 0KUhKOQVQ 6 ÿþ999ý &KUTWRVKQP QH VhG YRB2 IGPG TGVCTFU PWENGCT RTQVGKP GZRQTV, ECWUKPI C RTQHQWPF OKVQVKE FGNC[, CPF ECP DG TGUEWGF D[ QXGTGZRTGUUKQP QH XPO1ðCRM1. J $KQEhGO þü5:57ô-585 0QTDGEM J, $NQODGTI # ÿþ998ý #OKPQ CEKF WRVCMG KU UVTQPIN[ CHHGEVGF FWTKPI GZRQPGPVKCN ITQYVh QH Saccharomyces cerevisiae KP û.7 / 0C%N OGFKWO. (©/ú /KETQDKQN õGVV þ58:þüþ-þüö 0QTDGEM J, 2ChNOCP #ò, #MhVCT 0, $NQODGTI #, #FNGT õ ÿþ99öý 2WTKHKECVKQP CPF EhCTCEVGTK\CVKQP QH VYQ KUQGP\[OGU QH &õ-IN[EGTQN-3-RhQURhCVCUG HTQO Saccharomyces cerevisiae. +FGPVKHKECVKQP QH VhG EQTTGURQPFKPI GPP1 CPF GPP2 IGPGU CPF GXKFGPEG HQT QUOQVKE TGIWNCVKQP QH )RRüR GZRTGUUKQP D[ VhG QUOQUGPUKPI OKVQIGP-CEVKXCVGF RTQVGKP MKPCUG UKIPCN VTCPUFWEVKQP RCVhYC[. J $KQN %hGO ü7þ:þ3875-þ388þ 1FC ó, 6QPQOWTC ò ÿþ993ý úGNGEVKQP QH C PQXGN DCMKPI UVTCKP HTQO VhG Torulaspora [GCUVU. $KQUEK $KQVGEh $KQEhGO 57:þ3üû-þ3üü 1FC ó, 6QPQOWTC ò ÿþ995ý ©NGEVTQRhQTGVKE MCT[QV[RKPI QH VhG [GCUV IGPWU Torulaspora. õGVV #RRN /KETQDKQN üþ:þ9û-þ93 1FC ó, óCDWMK /, 6QPQOWTC ò, (WMWPCIC / ÿþ997ý # Rh[NQIGPGVKE CPCN[UKU QH Saccharomyces URGEKGU D[ VhG UGSWGPEG QH þ8ú-ü8ú T40# URCEGT TGIKQPU. óGCUV þ3:þüô3þü5û 1NUUQP õ, 0KGNUGP J ÿüûûûý 6hG TQNG QH OGVCDQNKE GPIKPGGTKPI KP VhG KORTQXGOGPV QH Saccharomyces cerevisiae: WVKNK\CVKQP QH KPFWUVTKCN OGFKC. ©P\[OG /KETQD 6GEhPQN üö:785-79ü 1'4QWTMG ú/, *GTUMQYKV\ +, 1'úhGC ©ò ÿüûûüý óGCUV IQ VhG YhQNG *1) HQT VhG h[RGTQUOQVKE TGURQPUG. 6TGPFU )GPGV þ8:ôû5-ôþü 1UKPIC ò#, $GWFGMGT 4(, XCP FGT 2NCCV J$, *QNNCPFGT J# ÿþ988ý 0GY [GCUV UVTCKPU RTQXKFKPI HQT CP GPhCPEGF TCVG QH HGTOGPVCVKQP QH UWICTU, C RTQEGUU VQ QDVCKP UWEh [GCUVU CPF VhG WUG QH VhGUG [GCUVU. ©WTQRGCP 2CVGPV û3ûöþû7 #ü. f\ECP ú ÿüûûüý 6YQ FKHHGTGPV UKIPCNU TGIWNCVG TGRTGUUKQP CPF KPFWEVKQP QH IGPG GZRTGUUKQP D[ INWEQUG. J $KQN %hGO ü77:ôö993-ôö997 f\ECP ú, JQhPUVQP / ÿþ999ý (WPEVKQP CPF TGIWNCVKQP QH [GCUV hGZQUG VTCPURQTVGTU. /KETQDKQN /QN $KQN 4GX ö3:55ô-5ö9 f\ECP ú, õGQPI 6, JQhPUVQP / ÿþ99öý 4IVþR QH Saccharomyces cerevisiae, C MG[ TGIWNCVQT QH INWEQUG-KPFWEGF IGPGU, KU DQVh CP CEVKXCVQT CPF C TGRTGUUQT QH VTCPUETKRVKQP. /QN %GNN $KQN þö:öôþ9-öôüö f\ECP ú, øCNNKGT õ), (NKEM Jú, %CTNUQP /, JQhPUVQP / ÿþ997ý ©ZRTGUUKQP QH VhG SUC2 IGPG QH Saccharomyces cerevisiae KU KPFWEGF D[ NQY NGXGNU QH INWEQUG. óGCUV þ3:þü7þ37
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
93
2CTM J-+, )TCPV %/, #VVHKGNF 2ø, &CYGU +÷ ÿþ997ý 6hG HTGG\G-VhCY UVTGUU TGURQPUG QH VhG [GCUV Saccharomyces cerevisiae KU ITQYVh RhCUG URGEKHKE CPF KU EQPVTQNNGF D[ PWVTKVKQPCN UVCVG XKC VhG RAS-E[ENKE #/2 UKIPCN VTCPUFWEVKQP RCVhYC[. #RRN ©PXKTQP /KETQDKQN ö3:38þ8-38üô 2CTM J-+, )TCPV %/, &CXKU /J, &CYGU +÷ ÿþ998ý 6hG E[VQRNCUOKE %W,íP UWRGTQZKFG FKUOWVCUG QH Saccharomyces cerevisiae KU TGSWKTGF HQT TGUKUVCPEG VQ HTGG\G-VhCY UVTGUU. J $KQN %hGO ü73:üü9üþ-üü9ü8 2CUEWCN-#hWKT #, úGTTCPQ 4, 2TQHV / ÿüûûþý 6hG úMQþR TGRTGUUQT CPF )EPôR CEVKXCVQT CPVCIQPKUVKECNN[ OQFWNCVG UVTGUU-TGIWNCVGF VTCPUETKRVKQP KP Saccharomyces cerevisiae. /QN %GNN $KQN üþ:þö-ü5 2GFGTUGP /$ ÿþ998ý 6hG WUG QH PWENGQVKFG UGSWGPEG RQN[OQTRhKUOU CPF &0# MCT[QV[RKPI KP VhG KFGPVKHKECVKQP QH DTGYGT•U [GCUV UVTCKPU CPF KP OKETQDKQNQIKECN EQPVTQN. +P: õKPUMGPU *(, JCEMUQP J( ÿGFUý. $GGT CPCN[UKU. /QFGTP OGVhQFU QH RNCPV CPCN[UKU. 0GY UGTKGU, XQN 7. úRTKPIGT-øGTNCI, $GTNKP *GKFGNDGTI 0GY óQTM, RR þ8û-þ9ô. 2G[QW-0FK //, ÷CVVU Jõ, $TQYUG J ÿüûûûý KFGPVKHKECVKQP CPF EhCTCEVGTK\CVKQP QH CP CPKOCN •þü HCVV[ CEKF FGUCVWTCUG IGPG D[ hGVGTQNQIQWU GZRTGUUKQP KP Saccharomyces cerevisiae. #TEh $KQEhGO $KQRh[U 37ö:399-ôû8 2hKNKRU J, *GTUMQYKV\ + ÿþ997ý 1UOQVKE DCNCPEG TGIWNCVGU EGNN HWUKQP FWTKPI OCVKPI KP Saccharomyces cerevisiae. J %GNN $KQN þ38:9öþ-97ô 2KCQ *õ, 2Kh ò6, õKO J*, òCPI ú), JKP J$, òKO ú*, *YCPI +. ÿþ999ý #P #TCDKFQRUKU )úò3ðUhCII[-NKMG IGPG VhCV EQORNGOGPVU [GCUV UCNV UVTGUU-UGPUKVKXG OWVCPVU KU KPFWEGF D[ 0C%N CPF CDUEKUKE CEKF. 2NCPV 2h[UKQN þþ9:þ5ü7-þ53ô 2QUCU (, %hCODGTU J4, *G[OCP J#, *QGHHNGT J2, FG 0CFCN ©, #TKwQ J ÿüûûûý 6hG VTCPUETKRVKQPCN TGURQPUG QH [GCUV VQ UCNKPG UVTGUU. J $KQN %hGO ü75:þ7üô9-þ7ü55 2TCGMGNV ï/, /GCEQEM 2# ÿþ99ûý *ú2þü, C PGY UOCNN hGCV UhQEM IGPG QH Saccharomyces cerevisiae: CPCN[UKU QH UVTWEVWTG, TGIWNCVKQP CPF HWPEVKQP. /QN )GP )GPGV üü3:97-þûö 2TKGVQ J#, #IWKNGTC J, 4CPFG\-)KN ( ÿüûû3ý )GPGVKE GPIKPGGTKPI QH DCMGT•U [GCUV: EhCNNGPIGU CPF QWVNQQM. +P: òCNKFCU ú, 2QOGVVQ #, 2CNK[CVh ) ÿGFUý (QQF DKQVGEhPQNQI[: UGEQPF GFKVKQP, TGXKUGF CPF GZRCPFGF. /CTEGN &GMMGT +PE., 0GY óQTM ÿKP RTGUUý. 2TKQT %, 2QVKGT ú, úQWEKGV Jõ, ú[EhTQXC * ÿþ99öý %hCTCEVGTK\CVKQP QH VhG NHA1 IGPG GPEQFKPI C 0Cîðòî-CPVKRQTVGT QH VhG [GCUV Saccharomyces cerevisiae. (©$ú õGVV 387:89-93 4CFLK /, òKO J-/, 6QICP 6, óQUhKMCYC *, úhKTChKIG ò ÿüûûþý 6hG ENQPKPI CPF EhCTCEVGTK\CVKQP QH VhG CDC50 IGPG HCOKN[ KP Saccharomyces cerevisiae. óGCUV þ8:þ95-üû5 4CPFG\-)KN (, úCP\ 2 ÿþ99ôý %QPUVTWEVKQP QH KPFWUVTKCN DCMGT•U [GCUV UVTCKPU CDNG VQ CUUKOKNCVG OCNVQUG WPFGT ECVCDQNKVG TGRTGUUKQP EQPFKVKQPU. #RRN /KETQDKQN $KQVGEhPQN ôü:58þ-58ö 4CPFG\-)KN (, $NCUEQ #, 2TKGVQ J#, úCP\ 2 ÿþ995ý &1)4þ CPF &1)4ü: VYQ IGPGU HTQO Saccharomyces cerevisiae VhCV EQPHGT ü-FGQZ[INWEQUG TGUKUVCPEG YhGP QXGTGZRTGUUGF. óGCUV þþ:þü33-þüôû 4CPFG\-)KN (, úCP\ 2, 2TKGVQ J# ÿþ999ý ©PIKPGGTKPI DCMGT•U [GCUV: TQQO HQT KORTQXGOGPV. 6TGPFU $KQVGEhPQN þ7:ü37-üôô 4CPFG\-)KN (, 2TKGVQ J#, *GTPCPFG\-õQRG\ /J ÿüûûüý ïVKNK\CVKQP QH Torulaspora delbrueckii UVTCKPU KP VhG RTQFWEVKQP QH UYGGV FQWIh. 2%6ð©úûüðûûô3þ 4GGF ), 0CIQFCYKVhCPC 6÷ ÿþ99þý óGCUV 6GEhPQNQI[ÿüPF GFPý. øCP 0QUVTCPF 4GKPhQNF. 4GKUGT ø, 4WKU *, #OOGTGT ) ÿþ999ý òKPCUG CEVKXKV[-FGRGPFGPV PWENGCT GZRQTV QRRQUGU UVTGUU-KPFWEGF PWENGCT CEEWOWNCVKQP CPF TGVGPVKQP QH *QIþ OKVQIGP-CEVKXCVGF RTQVGKP MKPCUG KP VhG DWFFKPI [GCUV Saccharomyces cerevisiae. /QN $KQN %GNN þû:þþô7-þþöþ
9ô
(. 4CPFG\-)KN GV CN.
4GOK\G (, 4QWUVCP Jõ, úCDNC[TQNNGU J/, $CTTG 2, &GSWKP ú ÿþ999ý )N[EGTQN QXGTRTQFWEVKQP D[ GPIKPGGTGF Saccharomyces cerevisiae YKPG [GCUV UVTCKPU NGCFU VQ UWDUVCPVKCN EhCPIGU KP D[-RTQFWEV HQTOCVKQP CPF VQ C UVKOWNCVKQP QH HGTOGPVCVKQP TCVG KP UVCVKQPCT[ RhCUG. #RRN ©PXKTQP /KETQDKQN ö5:þô3-þô9 4GOK\G (, $CTPCXQP õ, &GSWKP ú ÿüûûþý )N[EGTQN GZRQTV CPF IN[EGTQN 3-RhQURhCVG FGh[FTQIGPCUG, DWV PQV IN[EGTQN RhQURhCVCUGU, CTG TCVG NKOKVKPI HQT IN[EGTQN RTQFWEVKQP KP Saccharomyces cerevisiae. /GVCD ©PI 3:3ûþ-3þü 4GR /, 4GKUGT ø, )CTVPGT ï, 6hGXGNGKP J/, *QhOCPP ú, #OOGTGT ), 4WKU * ÿþ999ý 1UOQVKE UVTGUU-KPFWEGF IGPG GZRTGUUKQP KP Saccharomyces cerevisiae TGSWKTGU /UPþR CPF VhG PQXGN PWENGCT HCEVQT *QVþR. /QN %GNN $KQN þ9:5ô7ô-5ô85 4GR /, òTCPV\ /, 6hGXGNGKP J/, *QhOCPP ú ÿüûûûý 6hG VTCPUETKRVKQPCN TGURQPUG QH Saccharomyces cerevisiae VQ QUOQVKE UhQEM. *QVþR CPF /UPüRð/UPôR CTG TGSWKTGF HQT VhG KPFWEVKQP QH UWDUGVU QH hKIh QUOQNCTKV[ IN[EGTQN RCVhYC[-FGRGPFGPV IGPGU. J $KQN %hGO ü75:8ü9û-83ûû 4KPExP #/, %QFxP #%, %CUVTGLxP (, $GPsVG\ 6 ÿüûûþý +ORTQXGF RTQRGTVKGU QH DCMGT•U [GCUV OWVCPVU TGUKUVCPV VQ ü-FGQZ[-&-INWEQUG. #RRN ©PXKTQP /KETQDKQN ö7:ôü79-ôü85 4KROCUVGT 6õ, øCWIhP )2, ÷QQNHQTF JT Jõ ÿþ993ý DRS1 VQ DRS7, PQXGN IGPGU TGSWKTGF HQT TKDQUQOG CUUGODN[ CPF HWPEVKQP KP Saccharomyces cerevisiae. /QN %GNN $KQN þ3:79ûþ79þü 4QFTKIWG\-0CXCTTQ # ÿüûûûý 2QVCUKWO VTCPURQTV KP HWPIK CPF RNCPVU. $KQEhKO $KQRh[U #EVC þôö9:þ-3û 4QFTKIWG\-øCTICU ú, ©UVTWEh (, 4CPFG\-)KN ( ÿüûûüý )GPG GZRTGUUKQP CPCN[UKU QH EQNF CPF HTGG\G UVTGUU KP DCMGT•U [GCUV. #RRN ©PXKTQP /KETQDKQN ö8:3ûüô-3û3û 4QIGTU &6, ú\QUVCM J÷ ÿþ987ý óGCUV UVTCKPU. 2%6 ÷187ðû3ûûö 4QNNCPF (, ÷KPFGTKEMZ J, 6hGXGNGKP J/ ÿüûûüý )NWEQUG-UGPUKPI CPF UKIPCNNKPI OGEhCPKUOU KP [GCUV. (©/ú óGCUV 4GU ü:þ83-üûþ 4QPPG * ÿþ995ý )NWEQUG TGRTGUUKQP KP HWPIK. 6TGPFU )GPGV þþ:þü-þ7 4WUPCM (, /GTV\ 2 ÿüûûûý %CNEKPGWTKP: HQTO CPF HWPEVKQP. 2h[UKQN 4GX 8û:þô83-þ5üþ úChCTC 6, )QFC 6, 1hIK[C ú ÿüûûüý %QORTGhGPUKXG GZRTGUUKQP CPCN[UKU QH VKOG-FGRGPFGPV IGPGVKE TGURQPUGU KP [GCUV EGNNU VQ NQY VGORGTCVWTG. J $KQN %hGO ü77:5ûûþ5-5ûûüþ úCMCOQVQ 6, /WTCVC 0 ÿüûûüý 4GIWNCVKQP QH VhG FGUCVWTCVKQP QH HCVV[ CEKFU CPF KVU TQNG KP VQNGTCPEG VQ EQNF CPF UCNV UVTGUU. %WTT 1RKP /KETQDKQN 5:üûö-üþû úCNGU ò, $TCPFV ÷, 4WODCM ©, õKPFUG[ ) ÿüûûûý 6hG õ©#-NKMG RTQVGKP *ú2þü KP Saccharomyces cerevisiae hCU C RNCUOC OGODTCPG NQECVKQP CPF RTQVGEVU OGODTCPGU CICKPUV FGUKEECVKQP CPF GVhCPQN-KPFWEGF UVTGUU. $KQEhKO $KQRh[U #EVC þôö3:üö7-ü78 úCOWGN &, òWOCT 6òú, )CPGUh ), JC[CTCOCP ), óCPI 2-÷, %hCPI /-/, 6TKXGFK ø&, ÷CPI ú-õ, *YCPI ò-%, %hCPI &-ò, óW % ÿüûûûý 2TQNKPG KPhKDKVU CIITGICVKQP FWTKPI RTQVGKP TGHQNFKPI. 2TQVGKP úEK 9:3ôô-35ü úCPQ (, #UCMCYC 0, +PQWG ó, úCMWTCK / ÿþ999ý # FWCN TQNG HQT KPVTCEGNNWNCT VTGhCNQUG KP VhG TGUKUVCPEG QH [GCUV EGNNU VQ YCVGT UVTGUU. %T[QDKQNQI[ 39:8û-87 úCP\ 2, 4CPFG\-)KN (, 2TKGVQ J# ÿþ99ôý /QNGEWNCT EhCTCEVGTK\CVKQP QH C IGPG VhCV EQPHGTU üFGQZ[INWEQUG TGUKUVCPEG KP [GCUV. óGCUV þû:þþ95-þüûü úCP\ 2, #NOU )4, *C[UVGCF 6#, %CTNUQP / ÿüûûûý 4GIWNCVQT[ KPVGTCEVKQPU DGVYGGP VhG 4GIþ-)NE7 RTQVGKP RhQURhCVCUG CPF VhG úPHþ RTQVGKP MKPCUG. /QN %GNN $KQN üû:þ3üþþ3ü8 úCUCMK 6, 1hUhKOC ó ÿþ987ý +PFWEVKQP CPF EhCTCEVGTK\CVKQP QH CTVKHKEKCN FKRNQKU HTQO VhG hCRNQNKF [GCUV Torulaspora delbrueckii. #RRN ©PXKTQP /KETQDKQN 53:þ5ûô-þ5þþ úEhOGN\NG 6, *CNN /0 ÿüûûûý 614, C EGPVTCN EQPVTQNNGT QH EGNN ITQYVh. %GNN þû3:ü53-üöü
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
95
úEhPGNN 0, òTGOU $, ©PVKCP ò& ÿþ99üý 6hG PAR1 ÿYAP1ðSNQ3ý IGPG QH Saccharomyces cerevisiae, C E-LWP hQOQNQIWG, KU KPXQNXGF KP QZ[IGP OGVCDQNKUO. %WTT )GPGV üþ:üö9ü73 úEhWNNGT &, %QTVG-4GCN /, õGCQ % ÿüûûþý # FKHHGTGPVKCN OGFKWO HQT VhG GPWOGTCVKQP QH VhG URQKNCIG [GCUV Zygosaccharomyces bailii KP YKPG. J (QQF 2TQV ö3:þ57û-þ575 úGTTCPQ 4 ÿþ99öý úCNV VQNGTCPEG KP RNCPVU CPF OKETQQTICPKUOU: VQZKEKV[ VCTIGVU CPF FGHGPUG TGURQPUGU. +PV 4GX %[VQN þö5:þ-5ü úGTTCPQ 4, 4QFTKIWG\-0CXCTTQ # ÿüûûþý +QP hQOGQUVCUKU FWTKPI UCNV UVTGUU KP RNCPVU. %WTT 1RKP %GNN $KQN þ3:399-ôûô úGTTCPQ 4, /áTSWG\ J#, 4sQU ) ÿþ997ý %TWEKCN HCEVQTU KP UCNV UVTGUU VQNGTCPEG. +P: *QhOCPP ú, /CIGT ÷* ÿGFUý óGCUV úVTGUU 4GURQPUGU. úRTKPIGT-øGTNCI, *GKFGNDGTI, RR þô7-þö9 úGTTCPQ 4, /WNGV J/, 4KQU ), /CTSWG\ J#, FG õ+(, õGWDG /2, /GPFK\CDCN +, 2CUEWCN ##, 2TQHV /, 4QU 4, /QPVGUKPQU % ÿþ999ý # INKORUG QH VhG OGEhCPKUOU QH KQP hQOGQUVCUKU FWTKPI UCNV UVTGUU. J ©ZR $QV 5û:þûü3-þû3ö úhGP $, *QhOCPP ú, JGPUGP 4), $QhPGTV * ÿþ999ý 4QNGU QH UWICT CNEQhQNU KP QUOQVKE UVTGUU CFCRVCVKQP. 4GRNCEGOGPV QH IN[EGTQN D[ OCPPKVQN CPF UQTDKVQN KP [GCUV. 2NCPV 2h[UKQN þüþ:ô5-5ü úhKOC J, *KPQ #, óCOCFC-+[Q %, úW\WMK ó, 0CMCLKOC 4, ÷CVCPCDG *, /QTK ò, 6CMCPQ * ÿþ999ý úVTGUU VQNGTCPEG KP FQWIhU QH Saccharomyces cerevisiae VTGhCNCUG OWVCPVU FGTKXGF HTQO EQOOGTEKCN DCMGT•U [GCUV. #RRN ©PXKTQP /KETQDKQN ö5:ü8ôþ-ü8ôö úhKOC J, úCMCVC-6UWFC ó, úW\WMK ó, 0CMCLKOC 4, ÷CVCPCDG *, òCYCOQVQ ú, 6CMCPQ * ÿüûû3ý &KUTWRVKQP QH VhG CAR1 IGPG GPEQFKPI CTIKPCUG GPhCPEGU HTGG\G VQNGTCPEG QH VhG EQOOGTEKCN DCMGT•U [GCUV Saccharomyces cerevisiae. #RRN ©PXKTQP /KETQDKQN ö9:7þ5-7þ8 úhKTC[COC /, /CVUWK ó, 6Qh-G # ÿþ99öý &QOKPCPV OWVCPV CNNGNGU QH [GCUV RTQVGKP MKPCUG IGPG CDC15 UWRRTGUU VhG NVGþ FGHGEV KP VGTOKPCVKQP QH / RhCUG CPF IGPGVKECNN[ KPVGTCEV YKVh CDC14. /QN )GP )GPGV ü5þ:þ7ö-þ85 úNGCVQT 4&, *KNN % ÿüûûþý $CEVGTKCN QUOQCFCRVCVKQP: VhG TQNG QH QUOQN[VGU KP DCEVGTKCN UVTGUU CPF XKTWNGPEG. (©/ú /KETQDKQN 4GX üö:ô9-7þ úOKVU *2, *CWH J, /WNNGT ú, *QDNG[ 6J, íKOOGTOCPP (ò, *ChP-*kIGTFCN $, 0KGNUGP J, 1NUUQP õ ÿüûûûý úKOWNVCPGQWU QXGTGZRTGUUKQP QH GP\[OGU QH NQYGT RCTV QH IN[EQN[UKU ECP GPhCPEG VhG HGTOGPVCVKXG ECRCEKV[ QH Saccharomyces cerevisiae.óGCUV þö:þ3ü5þ33ô úVCVhQRQWNQU #/, %[GTV /ú ÿþ997ý %CNEKPGWTKP CEVU VhTQWIh VhG CRZ1/TCN1-GPEQFGF VTCPUETKRVKQP HCEVQT VQ TGIWNCVG IGPG GZRTGUUKQP KP [GCUV. )GPGU &GX þþ:3ô3ü-3ôôô úVGTPGT &©, /QQP õGG J, *CTFKP ú©, )TGGPNGCH #õ ÿþ995ý 6hG [GCUV ECTDQZ[N-VGTOKPCN TGRGCV FQOCKP MKPCUG %6&ò-+ KU C FKXGTIGPV E[ENKP-E[ENKP-FGRGPFGPV MKPCUG EQORNGZ. /QN %GNN $KQN þ5:57þö-57üô úWVhGTNCPF (%÷, õCIGU (, õWECU %, õW[VGP ò, #NDGTV[P J, *QhOCPP ú, 2TKQT $#, òKNKCP ú) ÿþ997ý %hCTCEVGTKUVKEU QH (RUþ-FGRGPFGPV CPF KPFGRGPFGPV IN[EGTQN VTCPURQTV KP Saccharomyces cerevisiae. J $CEVGTKQN þ79:779û-7795 úYCP 6/, ÷CVUQP ò ÿþ998ý úVTGUU VQNGTCPEG KP C [GCUV UVGTQN CWZQVTQRh: TQNG QH GTIQUVGTQN, hGCV UhQEM RTQVGKPU CPF VTGhCNQUG. (©/ú /KETQDKQN õGVV þö9:þ9þ-þ97 úYKTG-%NCTM )#, /CTEQVVG JT ÷4 ÿþ999ý 6hG YhGCV õ©# RTQVGKP ©O HWPEVKQPU CU CP QUOQRTQVGEVKXG OQNGEWNG KP Saccharomyces cerevisiae. 2NCPV /QN $KQN 39:þþ7-þü8 6CQ ÷, &GUEhGPGU 4J, (CUUNGT Jú ÿþ999ý +PVTCEGNNWNCT IN[EGTQN NGXGNU OQFWNCVG VhG CEVKXKV[ QH úNPþR, C Saccharomyces cerevisiae VYQ-EQORQPGPV TGIWNCVQT. J $KQN %hGO ü7ô:3öû3ö7
9ö
(. 4CPFG\-)KN GV CN.
6CMCIK *, +YCOQVQ (, 0CMCOQTK ú ÿþ997ý +UQNCVKQP QH HTGG\G-VQNGTCPV NCDQTCVQT[ UVTCKP QH Saccharomyces cerevisiae HTQO RTQNKPG-CPCNQIWG-TGUKUVCPV OWVCPVU. #RRN /KETQDKQN $KQVGEhPQN ô7:ôû5-ôþþ 6CMCIK *, úCMCK ò, /QTKFC ò, 0CMCOQTK ú ÿüûûûý 2TQNKPG CEEWOWNCVKQP D[ OWVCVKQP QT FKUTWRVKQP QH VhG RTQNKPG QZKFCUG IGPG KORTQXGU TGUKUVCPEG VQ HTGG\KPI CPF FGUKEECVKQP UVTGUUGU KP Saccharomyces cerevisiae. (©/ú /KETQDKQN õGVV þ8ô:þû3-þû8 6COáU /J, õW[VGP ò, úWVhGTNCPF (%÷, *GTPCPFG\ #, #NDGTV[P, J, øCNCFK *, õK *, 2TKQT $#, òKNKCP ú), 4COQU J, )WUVCHUUQP õ, 6hGXGNGKP J/, *QhOCPP ú ÿþ999ý (RUþR EQPVTQNU VhG CEEWOWNCVKQP CPF TGNGCUG QH VhG EQORCVKDNG UQNWVG IN[EGTQN KP [GCUV QUOQTGIWNCVKQP. /QN /KETQDKQN 3þ:þû87-þþûô 6CPIhG #, 6GWPKUUGP #, øCP &KLEM 2, 6hGXGNGKP J/ ÿüûûûý +FGPVKHKECVKQP QH IGPGU TGURQPUKDNG HQT KORTQXGF ET[QTGUKUVCPEG KP HGTOGPVKPI [GCUV EGNNU. +PV J (QQF /KETQDKQN 55:ü59-üöü 6CPIhG #, øCP &KLEM 2, &WOQTVKGT (, 6GWPKUUGP #, *QhOCPP ú, 6hGXGNGKP J/ ÿüûûüý #SWCRQTKP GZRTGUUKQP EQTTGNCVGU YKVh HTGG\G VQNGTCPEG KP DCMGT•U [GCUV, CPF QXGTGZRTGUUKQP KORTQXGU HTGG\G VQNGTCPEG KP KPFWUVTKCN UVTCKPU. #RRN ©PXKTQP /KETQDKQN ö8:598þ5989 6GWPKUUGP #, &WOQTVKGT (, )QTYC /-(, $CWGT J, 6CPIhG #, õQvG\ #, úOGV 2, øCP &KLEM 2, 6hGXGNGKP J/ ÿüûûüý +UQNCVKQP CPF EhCTCEVGTK\CVKQP QH C HTGG\G-VQNGTCPV FKRNQKF FGTKXCVKXG QH CP KPFWUVTKCN DCMGT•U [GCUV UVTCKP CPF KVU WUG KP HTQ\GP FQWIhU. #RRN ©PXKTQP /KETQDKQN ö8:ô78û-ô787 6hCOOCXQPIU $, 2CPQHH J-/, )WoIWGP / ÿüûûûý 2hGPQV[RKE CFCRVCVKQP VQ HTGG\G-VhCY UVTGUU QH VhG [GCUV-NKMG HWPIWU Geotrichum candidum. +PV J (QQF /KETQDKQN öû:99-þû5 6hGXGNGKP J/, *QhOCPP ú ÿþ995ý 6TGhCNQUG U[PVhCUG, IWCTF VQ VhG ICVG QH IN[EQN[UKU KP [GCUV! 6TGPFU $KQEhGO úEK üû:3-þû 6hGXGNGKP J/, FG ÷KPFG J* ÿþ999ý 0QXGN UGPUKPI OGEhCPKUOU CPF VCTIGVU HQT VhG E#/2RTQVGKP MKPCUG # RCVhYC[ KP VhG [GCUV Saccharomyces cerevisiae. /QN /KETQDKQN 33:9ûô-9þ8 6hKGTKPIGT *#, JQPGU 2), +PQW[G / ÿþ998ý %QNF UhQEM CPF CFCRVCVKQP. $KQ©UUC[U üû:ô957 6UWLKOQVQ ó, +\CYC ú, +PQWG ó ÿüûûûý %QQRGTCVKXG TGIWNCVKQP QH DOG2, GPEQFKPI üFGQZ[INWEQUG-ö-RhQURhCVG RhQURhCVCUG, D[ úPHþ MKPCUG CPF VhG hKIh-QUOQNCTKV[ IN[EGTQN-OKVQIGP-CEVKXCVGF RTQVGKP MKPCUG ECUECFG KP UVTGUU TGURQPUGU QH Saccharomyces cerevisiae. J $CEVGTKQN þ8ü:5þüþ-5þüö øCNGPVKPG Jú, ÷GTV\ &õ, õ[QPU 6J, õKQW õ-õ, )QVQ JJ, )TCNNC ©$ ÿþ998ý 6hG FCTM UKFG QH FKQZ[IGP DKQEhGOKUVT[. %WTT 1RKP %hGO $KQN ü:ü53-üöü XCP &KLEM 2, %QNCXK\\C &, úOGV 2, 6hGXGNGKP J/ ÿþ995ý &KHHGTGPVKCN KORQTVCPEG QH VTGhCNQUG KP UVTGUU TGUKUVCPEG KP HGTOGPVKPI CPF PQPHGTOGPVKPI Saccharomyces cerevisiae EGNNU. #RRN ©PXKTQP /KETQDKQN öþ:þû9-þþ5 XCP &KLEM 2, /C 2, øGTUGNG /, )QTYC /-(, %QNQODQ ú, õGOCKTG ò, $QUUK &, õQvG\ #, 6hGXGNGKP J/ ÿüûûûý # DCMGT•U [GCUV OWVCPV ÿfil1ý YKVh C URGEKHKE, RCTVKCNN[ KPCEVKXCVKPI OWVCVKQP KP CFGP[NCVG E[ENCUG OCKPVCKPU C hKIh UVTGUU TGUKUVCPEG FWTKPI CEVKXG HGTOGPVCVKQP CPF ITQYVh. J /QN /KETQDKQN $KQVGEhPQN ü:5üþ-53û XCP *GGUYKLM ÷%, $CMMGT $/, 6GWUKPM $, òhQNQFGPMQ $0, úQOUGP 1J, úPQGR Jõ, ÷GUVGThQHH *ø ÿþ999ý õKXG EQPVTQN QH VhG NKXKPI EGNN. $KQEhGO úQE 6TCPU ü7:üöþ-üöô XCP *QGM 2, XCP &KLMGP J2, 2TQPM J6 ÿüûûûý 4GIWNCVKQP QH HGTOGPVCVKXG ECRCEKV[ CPF NGXGNU QH IN[EQN[VKE GP\[OGU KP EhGOQUVCV EWNVWTGU QH Saccharomyces cerevisiae. ©P\[OG /KETQD 6GEhPQN üö:7üô-73ö
3 $CMGT•U [GCUV: EhCNNGPIGU CPF HWVWTG RTQURGEVU
97
XCP 4QQKLGP 4J, úEhQRRKPM 2J, $CCPMTGKU 4 ÿþ99ôý +ORTQXGOGPV QH ICU CPF CNEQhQN RTQFWEVKQP D[ [GCUV. ©WTQRGCP RCVGPV ©2ûöô5û9ô$þ øCTGNC J%, 2TCGMGNV ï/, /GCEQEM 2#, 2NCPVC 4J, /CIGT ÷* ÿþ995ý 6hG Saccharomyces cerevisiae HSP12 IGPG KU CEVKXCVGF D[ VhG hKIh-QUOQNCTKV[ IN[EGTQN RCVhYC[ CPF PGICVKXGN[ TGIWNCVGF D[ RTQVGKP MKPCUG #. /QN %GNN $KQN þ5:öü3ü-öüô5 øCWIhCP-/CTVKP #, /CTVKPK # ÿþ998ý Saccharomyces /G[GP GZ 4GGUU. +P òWTV\OCP %2, (GNN J÷ ÿGFUý 6hG óGCUVU. # VCZQPQO[E UVWF[. ©NUGXKGT úEKGPEG $.ø., #OUVGTFCO, RR 358-37þ ÷CPI íJ, 2QPVG J) JT ÿþ995ý úVQTCIG UVCDKNKV[ QH INWVGP-HQTVKHKGF HTQ\GP FQWIh. %GTGCN (QQFU ÷QTNF ôû:8ü7-83þ ÷GTPGT-÷CUhDWTPG /, $TCWP ©õ, JQhPUVQP )%, úKPIGT 4# ÿþ993ý úVCVKQPCT[ RhCUG KP Saccharomyces cerevisiae. /KETQDKQN 4GX 57:383-ôûþ ÷hKVUKVV /ú, %QNNKPU 4), /WNNGV J© ÿþ997ý /QFWNCVKQP QH FGh[FTCVKQP VQNGTCPEG KP UQ[DGCP UGGFNKPIU ÿ&Gh[FTKP /CVþ KU KPFWEGF D[ FGh[FTCVKQP DWV PQV D[ CDUEKUKE CEKFý. 2NCPV 2h[UKQN þþô:9þ7-9ü5 ÷KP\GNGT ©#, úhQGOCMGT &&, #UVTQOQHH #, õKCPI *, #PFGTUQP ò, #PFTG $, $CPIhCO 4, $GPKVQ 4, $QGMG J&, $WUUG[ *, %hW #/, %QPPGNN[ %, &CXKU ò, &KGVTKEh (, &QY ú÷, ©N $CMMQWT[ /, (QWT[ (, (TKGPF ú*, )GPVCNGP ©, )KCGXGT ), *GIGOCPP J*, JQPGU 6, õCWD /, õKCQ *, &CXKU 4÷, GV CN. ÿþ999ý (WPEVKQPCN EhCTCEVGTK\CVKQP QH VhG S. cerevisiae IGPQOG D[ IGPG FGNGVKQP CPF RCTCNNGN CPCN[UKU. úEKGPEG ü85:9ûþ-9ûö ÷QNHG J, $T[CPV ) ÿþ999ý (TGG\KPI, ET[KPI, CPFðQT XKVTKHKECVKQP QH OGODTCPG-UQNWVG-YCVGT U[UVGOU. %T[QDKQNQI[ 39:þû3-þü9 ñW &, &WCP ñ, ÷CPI $, *QPI $, *Q 6*&, ÷W 4 ÿþ99öý ©ZRTGUUKQP QH C NCVG GODT[QIGPGUKU CDWPFCPV RTQVGKP IGPG, HVA1, HTQO DCTNG[ EQPHGTU VQNGTCPEG VQ YCVGT FGHKEKV CPF UCNV UVTGUU KP VTCPUIGPKE TKEG. 2NCPV 2h[UKQN þþû:üô9-ü57 óCNG J, $QhPGTV *J ÿüûûþý 6TCPUETKRV GZRTGUUKQP KP Saccharomyces cerevisiae CV hKIh UCNKPKV[. J $KQN %hGO ü7ö:þ599ö-þöûû7 óQUhKOQVQ *, úCNVUOCP ò, )CUEh #2, õK *ñ, 1ICYC 0, $QVUVGKP &, $TQYP 21, %[GTV /ú ÿüûûüý )GPQOG-YKFG CPCN[UKU QH IGPG GZRTGUUKQP TGIWNCVGF D[ VhG ECNEKPGWTKPð%T\þR UKIPCNKPI RCVhYC[ KP Saccharomyces cerevisiae. J $KQN %hGO ü77:3þû79-3þû88 íhCPI õ, óW õ, óW %-# ÿþ998ý )GPGTCVKQP QH úWRGTQZKFG #PKQP D[ úWEEKPCVG-%[VQEhTQOG c 4GFWEVCUG HTQO $QXKPG *GCTV /KVQEhQPFTKC. J $KQN %hGO ü73:3397ü-3397ö íhCPI õ, 1hVC #, 6CMCIK /, +OCK 4 ÿüûûûý ©ZRTGUUKQP QH RNCPV ITQWR ü CPF ITQWR 3 lea IGPGU KP Saccharomyces cerevisiae TGXGCNGF HWPEVKQPCN FKXGTIGPEG COQPI õ©# RTQVGKPU. J $KQEhGO þü7:öþþ-öþö íhCPI õ, 1hVC #, *QTKWEhK *, 6CMCIK /, +OCK 4 ÿüûûþý /WNVKRNG OGEhCPKUOU TGIWNCVG GZRTGUUKQP QH NQY VGORGTCVWTG TGURQPUKXG ÿLOTý IGPGU KP Saccharomyces cerevisiae. $KQEhGO $KQRh[U 4GU %QOOWP ü83:53þ-535
4 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU +UCM 5. 2TGVQTKWU 6JG #WUVTCNKCP 9KPG 4GUGCTEJ +PUVKVWVG, #FGNCKFG, #WUVTCNKC
#DUVTCEV #P WTIGPV PGGF JCU CTKUGP VQ FGXGNQR UVCTVGT EWNVWTG UVTCKPU QH Saccharomyces cerevisiae RQUUGUUKPI C YKFG TCPIG QH URGEKCNKUGF RTQRGTVKGU KP QTFGT VQ OGGV VJG PGY CPF EJCNNGPIKPI FGOCPFU QH VJG XCTKQWU YKPG RTQFWEGTU CPF EQPUWOGTU. 5VTCKP FGXGNQROGPV KU PQ NQPIGT NKOKVGF VQ VJG RTKOCT[ TQNG QH YKPG [GCUVU, PCOGN[ VQ ECVCN[UG VJG TCRKF CPF EQORNGVG EQPXGTUKQP QH ITCRG UWICTU VQ CNEQJQN CPF ECTDQP FKQZKFG YKVJQWV FKUVQTVKPI VJG HNCXQWT QH VJG HKPCN RTQFWEV. 6QFC[, VJGTG KU C OWEJ UVTQPIGT GORJCUKU QP VJG FGXGNQROGPV QH YKPG [GCUVU HQT VJG EQUV-GHHGEVKXG RTQFWEVKQP QH YKPG YKVJ OKPKOKUGF TGUQWTEG KPRWVU, KORTQXGF SWCNKV[ CPF NQY GPXKTQPOGPVCN KORCEV. 6JKU EJCRVGT HQEWUGU QP VJG IGPGVKE EQPUVKVWVKQP, CPCN[UKU, CPF KORTQXGOGPV QH YKPG [GCUVU CPF VJG RQVGPVKCN TQNG VJCV EWUVQOKUGF UVCTVGT [GCUV UVTCKPU EQWNF RNC[ KP KORTQXKPI VJG HGTOGPVCVKQP, RTQEGUUKPI CPF DKQRTGUGTXCVKQP QH YKPGU, VJGKT ECRCEKV[ VQ GPJCPEG VJG YJQNGUQOGPGUU CPF UGPUQT[ SWCNKV[ QH YKPG, CPF VJGKT EWTTGPV UVCVWU CPF HWVWTG.
4.1 +PVTQFWEVKQP 9KPG KU QHVGP TGHGTTGF VQ CU DQVVNGF RQGVT[; KV KU C WPKSWG EQOOQFKV[ VJCV RTGFCVGU TGEQTFGF JKUVQT[. 9KPG RTQFWEVKQP TGSWKTGU CP KPVGITCVKQP QH UEKGPEG, GEQPQOKEU CPF CTVKUVT[ - C DNGPF QH KPFKXKFWCN ETGCVKXGPGUU CPF KPPQXCVKXG VGEJPQNQI[. 6JG CVVTCEVKQP QH YKPG FQGU PQV TGN[ WRQP DQNF, EQPUKUVGPV HNCXQWTU, DWV TCVJGT WRQP C UWDVNG CUUQTVOGPV QH EJCPIKPI UGPUCVKQPU VJCV OCMG KVU EJCTOKPI TQOCPVKEKUO FKHHKEWNV VQ FGHKPG ($KUUQP GV CN. ÿ00ÿ). 7PVKN VJG UGEQPF JCNH QH VJG RTGXKQWU EGPVWT[, KV YCU RTKOCTKN[ VJG RTQFWEGT YJQ FGHKPGF YKPG SWCNKV[; VJG OQFGTP INQDCNKUGF OCTMGV JCU EJCPIGF VJKU, CPF VJG EQPUWOGT PQY FGEKFGU (2TGVQTKWU CPF $CWGT ÿ00ÿ). /CTMGVKPI C UGPUQT[ GZRGTKGPEG KP VJG ÿ1UV EGPVWT[, VJGTGHQTG, KU KPETGCUKPIN[ FGRGPFGPV WRQP C DGVVGT WPFGTUVCPFKPI QH VJG DCUKE DKQNQI[ QH JWOCP RGTEGRVKQP CPF QNHCEVQT[ CPF HNCXQWT RTGHGTGPEGU. #PF KP CFFKVKQP VQ FGOCPFKPI C RTQFWEV VJCV KU GPLQ[CDNG KP CNN UGPUQT[ CURGEVU, VQFC['U EQPUWOGTU KPETGCUKPIN[ Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
100
+UCM 5. 2TGVQTKWU
GZRGEV YKPGU VQ DG JGCNVJHWN CPF RTQFWEGF KP CP GPXKTQPOGPVCNN[ UWUVCKPCDNG OCPPGT ($KUUQP GV CN. ÿ00ÿ). 6JG EJCNNGPIG, VJGTGHQTG, HCEKPI VJG YKPG RTQFWEGT VQFC[ JCU CV NGCUV VJTGG EQORQPGPVU: VQ WPFGTUVCPF YJ[ EQPUWOGTU OCMG VJG EJQKEGU VJG[ FQ, VQ RTQXKFG RTQFWEVU VJCV UCVKUH[ VJQUG EJQKEGU, CPF VQ GORNQ[ UWUVCKPCDNG RTQFWEVKQP RTCEVKEGU HQT ITQYKPI VJG ITCRGU CPF OCMKPI VJG YKPG ($KUUQP GV CN. ÿ00ÿ; 2TGVQTKWU CPF $CWGT ÿ00ÿ; 8KXKGT CPF 2TGVQTKWU ÿ00ÿ). 6JG UVCMGU CTG JKIJ: YKPG GZRQTVU CPF YKPG-TGNCVGF VQWTKUO CTG OCLQT EQPVTKDWVQTU VQ VJG GEQPQOKGU QH OCP[ YKPGRTQFWEKPI TGIKQPU CPF EQWPVTKGU. ©PJCPEGF VGEJPQNQI[ YKNN DG ETWEKCN VQ VJG UWEEGUU QH VJG KPVGTPCVKQPCN YKPG KPFWUVT[; VJKU KPENWFGU DKQVGEJPQNQIKECN KPPQXCVKQP CKOGF CV QXGTEQOKPI YKPG KPFWUVT[ RTQDNGOU CPF ETGCVKPI PGY QRRQTVWPKVKGU. &GURKVG XGT[ XQECN QRRQUKVKQP VQ IGPGVKECNN[ OQFKHKGF QTICPKUOU (þ/ýU) CPF RTQFWEVU, VJG INQDCN YKPG KPFWUVT[ KPETGCUKPIN[ KU HQEWUUKPI QP IGPGVKE KORTQXGOGPV QH VJG ITCRGXKPG CPF VJG YKPG [GCUV, VJG VYQ OCKP QTICPKUOU KPXQNXGF KP VJG RTQFWEVKQP QH YKPG (HQT TGXKGYU UGG 2TGVQTKWU CPF 8CP FGT 9GUVJWK\GP 1üü1; $CTTG GV CN. 1üü3; %QNCITCPFG GV CN. 1üüû; úWGTQN CPF 4COxP 1üüù; *GPUEJMG 1üüø; þTQUUOCPP CPF 2TGVQTKWU 1üüü; ·UVGTICCTF GV CN. ÿ000; 2TGVQTKWU 1üüü, ÿ000; 4CKPKGTK CPF 2TGVQTKWU ÿ000; &GSWKP ÿ001; 2TGVQTKWU ÿ001; 2TGVQTKWU CPF $CWGT ÿ00ÿ; 8KXKGT CPF 2TGVQTKWU ÿ000, ÿ00ÿ; 2TGVQTKWU ÿ003; 2TGVQTKWU GV CN. ÿ003). 6JKU EJCRVGT CKOU VQ U[PVJGUKUG KPHQTOCVKQP HTQO VJG NKVGTCVWTG QP VJG IGPGVKE EQPUVKVWVKQP, CPCN[UKU CPF KORTQXGOGPV QH YKPG [GCUVU. 6JG EJCRVGT CNUQ FKUEWUUGU VJG TQNG EWUVQOKUGF UVCTVGT [GCUV UVTCKPU EQWNF RNC[ KP KORTQXKPI HGTOGPVCVKQP, RTQEGUUKPI CPF DKQRTGUGTXCVKQP QH YKPGU, VJGKT ECRCEKV[ VQ GPJCPEG YJQNGUQOGPGUU CPF UGPUQT[ SWCNKV[, CPF VJGKT EWTTGPV UVCVWU CPF HWVWTG.
4.2 6JG PGGF HQT KORTQXGF YKPG [GCUV UVTCKPU 4.2.1 6JG CFXCPVCIGU CPF FKUCFXCPVCIGU QH URQPVCPGQWU CPF KPQEWNCVGF HGTOGPVCVKQPU 6JG [GCUVU HQWPF QP VJG UWTHCEGU QH ITCRGU CPF YKPGT[ GSWKROGPV JCXG JKUVQTKECNN[ DGGP TGURQPUKDNG HQT VJG EQPXGTUKQP QH ITCRG LWKEG KPVQ YKPG. +P VJGUG URQPVCPGQWU HGTOGPVCVKQPU, VJGTG KU C UGSWGPVKCN ITQYVJ RCVVGTP QH KPFKIGPQWU [GCUVU (G.I., Kloeckera/Hanseniaspora, Candida, Metschnikowia, Pichia, GVE.), YKVJ VJG HKPCN UVCIGU QH HGTOGPVCVKQP FQOKPCVGF D[ VJG CNEQJQN-VQNGTCPV UVTCKPU QH Saccharomyces cerevisiae (2TGVQTKWU GV CN. 1üüü). 6JKU JCU GCTPGF S. cerevisiae VJG FGUKIPCVKQP QH ÉVJG YKPG [GCUVÌ, CPF KP VJG NCUV HQWT QT HKXG FGECFGU, KPETGCUGF WPFGTUVCPFKPI QH VJG FKXGTUKV[, RJ[UKQNQI[ CPF IGPGVKEU QH S. cerevisiae UVTCKPU JCU NGF VQ VJG UGNGEVKQP QH UVCTVGT EWNVWTG UVTCKPU HQT VJGKT HGTOGPVCVKQP DGJCXKQWT. #NVJQWIJ URQPVCPGQWU HGTOGPVCVKQPU QH ITCRG LWKEG TGUWNV KP JKIJN[ WPRTGFKEVCDNG QWVEQOGU CPF WUWCNN[ VCMG OWEJ NQPIGT VJCP OQUV YKPGOCMGTU CTG YKNNKPI VQ CFOKV, OCP[ YKPGTKGU UVTKXKPI HQT XKPVCIG XCTKCDKNKV[ CEEGRV VJGUG TKUMU CU VJG RTKEG QH CEJKGXKPI FKUVKPEV UV[NGU QH YKPG (*GPUEJMG 1üüø). 6JGUG YKPGOCMGTU DGNKGXG
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
101
VJCV KPFKIGPQWU [GCUVU RTGUGPV KP URQPVCPGQWU YKPG HGTOGPVCVKQPU RTQFWEG JKIJGT EQPEGPVTCVKQPU QH IN[EGTQN CPF QVJGT RQN[QNU, [KGNFKPI YKPGU YKVJ C FKUVKPEV UGPUQTKCN SWCNKV[ QHVGP FGUETKDGF CU C HWNNGT, TQWPFGT RCNCVG UVTWEVWTG (÷WIGNUCPI 1üüø). ÷WTVJGTOQTG, URQPVCPGQWU HGTOGPVCVKQP ECWUGU CP GZVGPFGF NCI RJCUG DGHQTG VJG QPUGV QH XKIQTQWU HGTOGPVCVKQP CEVKXKV[, CNNQYKPI QZ[IGP VQ TGCEV YKVJ CPVJQE[CPKPU CPF QVJGT RJGPQNU KP VJG CDUGPEG QH GVJCPQN. 6JKU RTQEGUU KU VJQWIJV VQ GPJCPEG EQNQWT UVCDKNKV[ KP TGF YKPGU CPF VQ CEEGNGTCVG RJGPQN RQN[OGTKUCVKQP (÷WIGNUCPI 1üüø). öCTIG OQFGTP YKPGTKGU TGSWKTG TCRKF CPF TGNKCDNG HGTOGPVCVKQPU HQT EQPUKUVGPV YKPG HNCXQWT CPF RTGFKEVCDNG SWCNKV[; VJGTGHQTG VJG[ WUG URGEKCNN[ UGNGEVGF UVCTVGT EWNVWTGU QH S. cerevisiae YKVJ MPQYP CDKNKV[ (*GPUEJMG 1üüø). 6JGUG RWTG EWNVWTGU CTG KPQEWNCVGF KPVQ VJG ITCRG OWUV, HQNNQYKPI VJG CRRNKECVKQP QH UWNRJWT FKQZKFG (5ýÿ), VQ UWRRTGUU VJG RTQNKHGTCVKQP QH VJG PCVWTCN OKETQHNQTC, VQ TGFWEG VJG NCI RJCUG CPF VQ EQORNGVG UWICT EQPXGTUKQP VQ CNEQJQN OWEJ HCUVGT VJCP KP WPKPQEWNCVGF HGTOGPVCVKQPU (2TGVQTKWU ÿ000). *QYGXGT, VJG WUG QH S. cerevisiae UVCTVGT EWNVWTGU FQGU PQV PGEGUUCTKN[ RTGXGPV VJG RTGUGPEG CPF CEVKXKV[ QH YKPGT[-CUUQEKCVGF [GCUV UVTCKPU (÷WIGNUCPI 1üüø). +V KU IGPGTCNN[ CEEGRVGF VJCV DQVJ URQPVCPGQWU CPF KPQEWNCVGF YKPG HGTOGPVCVKQPU CTG CHHGEVGF D[ VJG FKXGTUKV[ QH [GCUVU CUUQEKCVGF YKVJ VJG XKPG[CTF CPF YKPGT[. 6JKU YGCNVJ QH [GCUV DKQFKXGTUKV[ KP VJG XCTKQWU YKPG-RTQFWEKPI TGIKQPU QH VJG YQTNF TGOCKPU NCTIGN[ WPVCRRGF HQT KVU QGPQNQIKECN RQVGPVKCN, NGCXKPI QRGP VJG RQUUKDKNKV[ VJCV C YKPGOCMGT OKIJV RTGHGT VQ WUG C OKZVWTG QH S. cerevisiae UVTCKPU CPF PQPSaccharomyces URGEKGU CU C UVCTVGT EWNVWTG. 5WEJ C OKZGF UVCTVGT EWNVWTG OKIJV GXGP DG VCKNQTGF VQ TGHNGEV VJG [GCUV DKQFKXGTUKV[ QH C IKXGP TGIKQP, TGUWNVKPI KP UGPUQT[ RTQRGTVKGU KP UWEEGUUKXG XKPVCIGU V[RKECN QH VJG YKPGU RTQFWEGF KP VJCV TGIKQP (2TGVQTKWU GV CN. 1üüü). 6JKU YQWNF EQTTGNCVG YKVJ VJG EQPEGRV QH VGTTQKT VJG KPHNWGPEG QH VJG NQECN GPXKTQPOGPV QP VJG EQORQUKVKQP QH YKPG RTQFWEGF KP C URGEKHKE ITQYKPI TGIKQP. 6JGTGHQTG, UQOG YKPGOCMGTU ENCKO VJCV VJG WUG QH C OKZVWTG QH NQECNN[ KUQNCVGF [GCUV URGEKGU YKVJ FGUKTCDNG QGPQNQIKECN RTQRGTVKGU YQWNF UWRRQTV VJG KFGC QH OKPKOCN KPVGTXGPVKQP UQ VJCV VJG VGTTQKT OC[ DG GXKFGPV KP VJG HKPCN RTQFWEV. 4.2.2 6JG FGXGNQROGPV QH CEVKXG FTKGF YKPG [GCUV UVCTVGT EWNVWTG UVTCKPU 6JG EQPEGRV QH UVCTVKPI YKPG HGTOGPVCVKQPU YKVJ RWTG [GCUV EWNVWTGU IQGU DCEM VQ 1õü0 YJGP /ôNNGT-6JWTICW EQPXKPEGF UQOG þGTOCP YKPGOCMGTU QH VJG DGPGHKVU QH VJG CUUQEKCVGF TCRKF CPF GXGP TCVGU QH HGTOGPVCVKQP. *QYGXGT, VJG NKOKVGF UWEEGUU QH VJG HKTUV VYQ EQOOGTEKCN CEVKXG FTKGF YKPG [GCUV UVTCKPU QHHGTGF KPVGTPCVKQPCNN[ CU CNN-RWTRQUG [GCUVU KP 1üùó OCFG KV ENGCT VJCV VJGTG YCU C PGGF HQT UGRCTCVG S. cerevisiae UVTCKPU YKVJ URGEKHKE EJCTCEVGTKUVKEU HQT FKHHGTGPV V[RGU CPF UV[NGU QH YKPG (&GITo 1üü3). 6JQWIJ YKPG [GCUV HWPEVKQPU RTKOCTKN[ VQ ECVCN[UG VJG GHHKEKGPV CPF EQORNGVG EQPXGTUKQP QH ITCRG UWICTU VQ CNEQJQN YKVJQWV VJG FGXGNQROGPV QH QHH-HNCXQWTU, VQFC[•U YKPGOCMGTU FGOCPF UVCTVGT EWNVWTG UVTCKPU YKVJ C YJQNG
10ÿ
+UCM 5. 2TGVQTKWU
(KI. 1. # UEJGOCVKE TGRTGUGPVCVKQP QH VJG E[VQNQI[ QH C YKPG [GCUV EGNN, CPF QH KVU PWENGWU CPF EJTQOQUQOGU, YJKEJ ECP DG UGRCTCVGF D[ RWNUGF HKGNF IGN GNGEVTQRJQTGUKU.
TCPIG QH URGEKCNKUGF RTQRGTVKGU. 6JG KORQTVCPEG QH VJGUG RTQRGTVKGU FKHHGTU YKVJ VJG V[RG CPF UV[NG QH YKPG VQ DG OCFG CPF VJG VGEJPKECN TGSWKTGOGPVU QH VJG YKPGT[ (2TGVQTKWU ÿ000), DWV KV KU ENGCT VJCV VJGTG KU C ITQYKPI PGGF HQT S. cerevisiae UVTCKPU VJCV CTG DGVVGT CFCRVGF VQ XCTKQWU YKPG ITCRG XCTKGVCNU, XKVKEWNVWTCN RTCEVKEGU CPF YKPGOCMKPI VGEJPKSWGU. 6JKU FGOCPF HQT URGEKCNKUGF YKPG [GCUV UVTCKPU JCU NGF VQ VJG FGXGNQROGPV QH TCRKF OQNGEWNCT OGVJQFU HQT YKPG [GCUV UVTCKP FKHHGTGPVKCVKQP. 6JG OGVJQFU OQUV HTGSWGPVN[ WUGF VQ FKUETKOKPCVG COQPI [GCUV UVTCKPU YKVJ UKOKNCT RJ[UKQNQIKECN EJCTCEVGTKUVKEU KPENWFG VJG HQNNQYKPI: EJTQOQUQOG NGPIVJ RQN[OQTRJKUOU (%ö2); TGUVTKEVKQP HTCIOGPV NGPIVJ RQN[OQTRJKUOU (4÷ö2); TCPFQON[ CORNKHKGF RQN[OQTRJKE &ò# (4#2&) WUKPI VJG RQN[OGTCUG EJCKP TGCEVKQP (2%4); CORNKHKGF HTCIOGPV NGPIVJ RQN[OQTRJKUOU (#÷ö2) WUKPI UVTCKP URGEKHKE 2%4 RTKOGTU; VGORGTCVWTG ITCFKGPV IGN GNGEVTQRJQTGUKU (6þþ©) QT FGPCVWTKPI ITCFKGPV IGN GNGEVTQRJQTGUKU (&þþ©) QH 2%4-CORNKHKGF RTQFWEVU; CPF ÷QWTKGT-VTCPUHQTO KPHTCTGF OKETQURGEVTQUEQR[. 6JG PGGF HQT FGFKECVGF UVTCKP FGXGNQROGPV RTQITCOOGU JCU CNUQ JKIJNKIJVGF VJG IGPGVKE HGCVWTGU QH YKPG [GCUVU CPF VJG OGVJQFU YKVJ YJKEJ VJG[ ECP DG CPCN[UGF CPF OQFKHKGF (2TGVQTKWU GV CN. 1üüü; 2TGVQTKWU ÿ000).
4.3 6JG IGPGVKE HGCVWTGU, CPCN[UKU CPF OQFKHKECVKQPU QH YKPG [GCUVU 4.3.1 6JG OQTRJQNQI[, TGRTQFWEVKQP CPF IGPGVKE EQPUVKVWVKQP QH YKPG [GCUVU +P IGPGTCN, YKPG [GCUV EGNNU JCXG CP GNNKRUQKFCN UJCRG (÷KI. 1) CPF TGRTQFWEG CUGZWCNN[ VJTQWIJ DWFFKPI (OKVQVKE FKXKUKQP), QT UGZWCNN[ D[ URQTWNCVKQP (OGKQVKE
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
103
UGITGICVKQP CPF JCRNQKFKUCVKQP) CPF OCVKPI (FKRNQKFKUCVKQP) (÷KI. ÿ). 6JG [GCUV PWENGWU KU C TQWPF-NQDCVG QTICPGNNG QH CDQWV 1.ó ±O KP FKCOGVGT, CPF C FQWDNG OGODTCPG EQPVCKPKPI RQTGU QH ÿ0-100 PO KP FKCOGVGT UGRCTCVGU VJG PWENGQRNCUO HTQO VJG E[VQRNCUO (÷KI. 1). 6JG PWENGCT OGODTCPGU CTG UQOGVKOGU EQPVKIWQWU YKVJ VJG GPFQRNCUOKE TGVKEWNWO CPF, WPNKMG OQUV GWMCT[QVKE EGNNU, VJG [GCUV PWENGCT OGODTCPG KU PQV FKUOCPVNGF FWTKPI OKVQUKU (9CNMGT 1üüõ). 6JG PWENGQRNCUO EQPVCKPU EJTQOCVKP, YJKEJ EQPUKUVU QH EQPFGPUGF DCUKE PWENGQRTQVGKP OCVGTKCN EQORTKUKPI FQWDNG-JGNKECN &ò#-JKUVQPG EQORNGZGU QTICPKUGF KP EJTQOQUQOGU. 6JG OCLQTKV[ QH NCDQTCVQT[ UVTCKPU QH S. cerevisiae CTG GKVJGT JCRNQKF QT FKRNQKF, YJGTGCU KPFWUVTKCN YKPG [GCUV UVTCKPU CTG RTGFQOKPCPVN[ FKRNQKF QT CPGWRNQKF, CPF QEECUKQPCNN[ RQN[RNQKF (5PQY 1üõ3). (÷QT OQTG FGVCKNU RNGCUG UGG %JCRVGT ÿ). +P IGPGTCN, S. cerevisiae KU EQPUKFGTGF VQ JCXG C TGNCVKXGN[ UOCNN IGPQOG, C NCTIG PWODGT QH EJTQOQUQOGU, NKVVNG TGRGVKVKXG &ò# CPF HGY KPVTQPU (þQHHGCW GV CN. 1üüù; ýNKXGT 1üüù). *CRNQKF UVTCKPU EQPVCKP 1ÿ VQ 13 OGICDCUGU (OD) QH PWENGCT &ò#, FKUVTKDWVGF CNQPI 1ù NKPGCT EJTQOQUQOGU. ©CEJ EJTQOQUQOG KU C UKPING &ò# OQNGEWNG YKVJ C NGPIVJ QH CDQWV ÿ00 VQ ÿÿ00 MKNQDCUGU (MD). +P CFFKVKQP VQ VJG EJTQOQUQOCN &ò#, C PWODGT QH PQP-/GPFGNKCP IGPGVKE GNGOGPVU CTG MPQYP VQ GZKUV KP VJG PWENGWU (G.I., 3ó VQ óó EQRKGU QH 6[ TGVTQVTCPURQUQPU RGT JCRNQKF IGPQOG CPF ó0 VQ 100 EQRKGU QH VJG ù.3 MD ÿËO RNCUOKF &ò#), OKVQEJQPFTKC (øó MD OV&ò#) CPF E[VQRNCUO (G.I., MKNNGT XKTCN-NKMG RCTVKENGU EQPVCKPKPI FU4ò# IGPQOGU, CPF RTKQP-NKMG GNGOGPVU UWEJ CU =2UK?, =©VC? CPF =74©ÿ?). *QYGXGT, RGT
(KI. 2. # UEJGOCVKE TGRTGUGPVCVKQP QH VJG EGNN E[ENG CPF NKHG E[ENG QH JGVGTQVJCNNKE CPF JQOQVJCNNKE YKPG [GCUV UVTCKPU.
10û
+UCM 5. 2TGVQTKWU
(KI. 3. # UEJGOCVKE QWVNKPG QH VGVTCF CPCN[UKU WUKPI C OKETQOCPKRWNCVQT. ÷QWT JCRNQKF CUEQURQTGU FGTKXGF HTQO C JGVGTQ\[IQVG FKRNQKF [GCUV EGNN (URA3/ura3) UJQY C /GPFGNKCP UGITGICVKQP RCVVGTP HQT VJG WTCEKN CNNGNGU, K.G., VYQ URA3: VYQ ura3
JCRU YKVJ VJG GZEGRVKQP QH VJG OV&ò#, VJGTG UGGOU VQ DG PQ GHHGEV QP VJG SWCNKV[ QH YKPG RTQFWEGF D[ S. cerevisiae UVTCKPU EQPVCKPKPI CP[ QH VJGUG PQPEJTQOQUQOCN IGPGVKE GNGOGPVU. 4.3.2 6JG IGPGVKE OGVJQFU HQT VJG CPCN[UKU CPF OQFKHKECVKQP QH YKPG [GCUVU ÷QT UGXGTCN FGECFGU, RQYGTHWN IGPGVKE OGVJQFU JCXG DGGP WUGF VQ CPCN[UG VJG /GPFGNKCP UGITGICVKQP RCVVGTPU QH URGEKHKE VTCKVU KP S. cerevisiae. +P RCTVKEWNCT, VGVTCF CPCN[UKU WUKPI C OKETQOCPKRWNCVQT JCU RTQXGF WUGHWN HQT VJG KPKVKCN KFGPVKHKECVKQP, EJCTCEVGTKUCVKQP CPF OCRRKPI QH DQVJ TGEGUUKXG CPF FQOKPCPV CNNGNGU QH S. cerevisiae (÷KI. 3). #HVGT VJG DKTVJ QH OQNGEWNCT IGPGVKE VGEJPKSWGU KP VJG 1üø0U CPF VJG CPPQWPEGOGPV QH VJG PWENGQVKFG UGSWGPEG QH VJG GPVKTG S. cerevisiae IGPQOG KP 1üüù, GXGP OQTG RQYGTHWN VQQNU JCXG DGGP FGXGNQRGF VQ CPCN[UG CPF OQFKH[ YKPG [GCUVU.
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
10ó
(KI. 4. # UEJGOCVKE TGRTGUGPVCVKQP QH VJG HNQY QH IGPGVKE KPHQTOCVKQP KP C YKPG [GCUV EGNN, CU YGNN CU VJG NGXGN QH CPCN[UKU, FGHKPKVKQP, UVCVWU CPF OGVJQF QH CPCN[UKU QH KVU IGPQOG VQ KVU VTCPUETKRVQOG, RTQVGQOG CPF OGVCDQNQOG.
6JG IGPQOKE UGSWGPEG JCU DGGP WUGF VQ FGUKIP CPF U[PVJGUKUG JKIJ-FGPUKV[ QNKIQPWENGQVKFG CTTC[U. 6JG CTTC[U KP VWTP OQPKVQT VJG NGXGNU QH GZRTGUUKQP QH PGCTN[ CNN VJG IGPGU QH [GCUV EGNNU ITQYP WPFGT C XCTKGV[ QH EWNVWTCN EQPFKVKQPU. ÷WTVJGTOQTG, UGXGTCN NCDQTCVQTKGU CTG U[UVGOCVKECNN[ UETGGPKPI FKUTWRVGF IGPGU KP FGNGVKQP (MPQEM-QWV) OWVCPVU VQ HWPEVKQPCNN[ CPCN[UG VJG [GCUV IGPQOG (ýNKXGT 1üüù). 6JG GXGPVWCN FGEKRJGTKPI QH VJG HWPEVKQPU QH S. cerevisiae•U ù000 IGPGU YKNN OCMG VJG EQORNGVG RTQVGQOG CEEGUUKDNG. &WTKPI RTQVGQOG CPCN[UKU, VJG RTQVGKP EQORNGOGPV KU FKURNC[GF CU URQVU QP C IGN VJCV ECP DG KFGPVKHKGF WUKPI RGRVKFG OCUU HKPIGTRTKPVKPI. ÷WTVJGTOQTG, OGVJQFU DGKPI FGXGNQRGF YKNN CNNQY VJG GXCNWCVKQP QH EGNNWNCT OGVCDQNKVGU CPF RQUUKDN[ VJG CPCN[UKU QH VJG GPVKTG OGVCDQNQOG. 6JG OGVJQFU HQT, CPF KPHQTOCVKQP CDQWV, VJG IGPQOG, VTCPUETKRVQOG, RTQVGQOG CPF OGVCDQNQOG QH C NCDQTCVQT[ UVTCKP QH S. cerevisiae ECP VJGP ITCFWCNN[ DG GZRCPFGF VQ VJG OWEJ OQTG EQORNGZ KPFWUVTKCN YKPG [GCUV UVTCKPU (÷KI. û). 9KPG [GCUVU ECP DG KORTQXGF IGPGVKECNN[ KP C PWODGT QH YC[U. 5QOG QH VJG OGVJQFU NKUVGF KP 6CDNG 1 CNVGT NKOKVGF TGIKQPU QH VJG IGPQOG, YJKNG QVJGTU CTG WUGF VQ TGEQODKPG QT TGCTTCPIG VJG GPVKTG IGPQOG (2TGVQTKWU ÿ000). 6JG ENCUUKE UGNGEVKQP CPF DTGGFKPI OGVJQFU (VJG UGNGEVKQP QH XCTKCPVU, OWVCPVU, J[DTKFU CPF E[VQFWEVCPVU) CTG XCNWCDNG HQT KORTQXKPI CPF EQODKPKPI VTCKVU WPFGT RQN[IGPKE EQPVTQN, DWV VJG[ CTG PQV URGEKHKE GPQWIJ VQ OQFKH[ C UKPING RTQRGTV[ YKVJQWV EQORTQOKUKPI QVJGT FGUKTGF EJCTCEVGTKUVKEU QH YKPG [GCUVU. (÷QT OQTG FGVCKNU RNGCUG UGG %JCRVGT ÿ).
10ù
+UCM 5. 2TGVQTKWU
6CDNG 1. )GPGVKE OGVJQFU HQT VJG KORTQXGOGPV QH YKPG [GCUV UVTCKPU /GVJQF
$TKGH FGUETKRVKQP QH VJG OGVJQF
#RRNKECVKQP
5GNGEVKQP QH XCTKCPVU
5KORNG CPF FKTGEV OGVJQF, DCUGF QP UGNGEVKQP QH PCVWTCN IGPGVKE XCTKCPVU HQWPF YKVJKP RQRWNCVKQPU QH CNN YKPG [GCUV UVTCKPU FWG VQ URQPVCPGQWU OWVCVKQPU (6[-RTQOQVGF EJTQOQUQOCN VTCPUNQECVKQP, OKVQVKE TGEQODKPCVKQP, IGPG EQPXGTUKQP, TGRNKECVKQP GTTQTU) #RRNKECVKQP QH OWVCIGPU VQ KPETGCUG VJG HTGSWGPE[ QH OWVCVKQPU KP C YKPG [GCUV RQRWNCVKQP. *KIJ HTGSWGPE[ QH OWVCVKQPU ECP NGCF VQ OWNVKRNG RJGPQV[RGU, YKVJ EGTVCKP VTCKVU DGKPI KORTQXGF, YJKNG QVJGTU CTG UKOWNVCPGQWUN[ FGDKNKVCVGF. 6JG GHHGEV QH RNQKF[ TGFWEGU GHHKEKGPE[ KP FKRNQKF QT RQN[RNQKF UVTCKPU. *CRNQKF UVTCKPU CTG RTGHGTTGF YJGP KPFWEKPI OWVCVKQPU. +PVTC-URGEKGU J[DTKFKUCVKQP GPVCKNU URQTWNCVKPI FKRNQKFU, TGEQXGTKPI KPFKXKFWCN JCRNQKF CUEQURQTGU CPF OCVKPI QH JCRNQKF EGNNU QH QRRQUKVG OCVKPI V[RGU VQ RTQFWEG C PGY JGVGTQ\[IQWU FKRNQKF. #U HQT CNN UGZWCN TGRTQFWEVKQP, VJG TGUWNVKPI FKRNQKF UVTCKPU OC[ UJQY RTQRGTVKGU VJCV CTG FKHHGTGPV HTQO VJCV QH GKVJGT RCTGPVCN UVTCKP. 6JGTG CTG UGXGTCN HQTOU QH URGEKCNKUGF V[RGU QH J[DTKFKUCVKQP KP ECUGU KP YJKEJ ENCUUKECN UGZWCN TGRTQFWEVKQP ECPPQV DG CRRNKGF. 6JGUG KPENWFG: 5RQTG-EGNN OCVKPI: /CP[ YKPG [GCUVU CTG JQOQVJCNNKE CPF TGSWKTG C FKTGEV URQTGEGNN OCVKPI RTQEGFWTG, YJKEJ GPVCKNU RNCEKPI HQWT JQOQVJCNNKE CUEQURQTGU HTQO VJG UCOG CUEWU KPVQ FKTGEV EQPVCEV YKVJ JGVGTQVJCNNKE JCRNQKF EGNNU D[ OKETQOCPKRWNCVKQP. /CVKPI YKNN QEEWT DGVYGGP EQORCVKDNG CUEQURQTGU CPF EGNNU. 4CTG OCVKPI: ÷QTEGF OCVKPI QH UVTCKPU VJCV FQ PQV GZRTGUU C OCVKPI V[RG. +PXQNXGU OKZKPI QH UVTCKPU CV JKIJ FGPUKVKGU CPF VJG CXCKNCDKNKV[ QH C UVTQPI, RQUKVKXG UGNGEVKQP RTQEGFWTG VQ UGNGEV TCTG J[DTKFU.
4GSWKTGU CXCKNCDKNKV[ QH CRRTQRTKCVG UGNGEVKQP RTQEGFWTGU (GPTKEJOGPVU, FKTGEV RQUKVKXG UGNGEVKQP, GVE.), UKPEG VJG PWODGT QH UKIPKHKECPV OWVCVKQP GXGPVU KU NQY. òQV WUGF HQT FTCOCVKE KORTQXGOGPVU.
/WVCIGPGUKU
*[DTKFKUCVKQP
6JG WUG QH OWVCIGPU (G.I., WNVTCXKQNGV NKIJV, GVJ[NOGVJCPG UWNRJQPCVG, òOGVJ[N, ò-PKVTQUQIWCPKFKPG) HQT FKTGEVGF UVTCKP FGXGNQROGPV KU NKOKVGF, DWV VJG RTQEGFWTG KU CRRNKGF VQ KUQNCVG PGY XCTKCPVU QH YKPG [GCUV UVTCKPU DGHQTG HWTVJGT IGPGVKE OCPKRWNCVKQP VCMGU RNCEG.
*[DTKFKUCVKQP KU VJG OQUV GHHGEVKXG OGVJQF HQT KORTQXKPI CPF EQODKPKPI VTCKVU WPFGT RQN[IGPKE EQPVTQN. 6JG KPENWUKQP QT GNKOKPCVKQP QH C URGEKHKE RTQRGTV[ ECP DG CEJKGXGF HCKTN[ SWKEMN[ D[ J[DTKFKUCVKQP, QP EQPFKVKQP VJCV VJG RTQRGTV[ JCU C UKORNG IGPGVKE DCUKU, HQT GZCORNG QPG QT VYQ IGPGU. *QYGXGT, OCP[ FGUKTCDNG YKPG [GCUV EJCTCEVGTKUVKEU CTG URGEKHKGF D[ UGXGTCN IGPGU QT CTG VJG TGUWNV QH UGXGTCN IGPG U[UVGOU KPVGTCEVKPI YKVJ QPG CPQVJGT. 5RQTG-EGNN OCVKPI KU WUGF VQ ETQUU JGVGTQVJCNNKE JCRNQKF UVTCKPU YKVJ VJG CUEQURQTGU QH JQOQVJCNNKE YKPG [GCUV UVTCKPU, VJGTGD[ EQODKPKPI RQN[IGPKE VTCKVU KPVQ C UKPING FKRNQKF J[DTKF UVTCKP. 6JG FKRNQKF YKNN DG JQOQVJCNNKE.
4CTG OCVKPI ECP DG WUGF VQ EQODKPG RQN[IGPKE VTCKVU HTQO VYQ PQP-OCVKPI FKRNQKF QT RQN[RNQKF UVTCKPU KPVQ C J[DTKF UVTCKP.
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
10ø
6CDNG 1. (EQPVKPWGF) /GVJQF
$TKGH FGUETKRVKQP QH VJG OGVJQF
#RRNKECVKQP
*[DTKFKUCVKQP
%[VQFWEVKQP: +PVTQFWEKPI E[VQRNCUOKE IGPGVKE GNGOGPVU KPVQ YKPG [GCUVU YKVJQWV VTCPUHGT QH PWENGCT IGPGU. %[VQFWEVKQP TGSWKTGU C JCRNQKF OCVKPI UVTCKP ECTT[KPI VJG kar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
%[VQFWEVKQP ECP DG WUGF VQ KPVTQFWEG IGPGVKE FGVGTOKPCPVU NQECVGF KP VJG E[VQRNCUO (MKNNGTñ\[OQEKP FU4ò#, OKVQEJQPFTKCN &ò#) CPF QVJGT RNCUOKFU.
6TCPUHQTOCVKQP
5RJGTQRNCUV HWUKQP KU WUGF VQ HWUG EGNNU YKVJ FKHHGTGPV NGXGNU QH RNQKF[. ÷QT GZCORNG, C FKRNQKF YKPG [GCUV UVTCKP ECP DG HWUGF VQ C JCRNQKF UVTCKP VQ IGPGTCVG VTKRNQKF UVTCKPU, QT VYQ FKRNQKF YKPG [GCUVU YKVJ EQORNGOGPVCT[ FGUKTCDNG VTCKVU ECP DG HWUGF VQ IGPGTCVG C VGVTCRNQKF YKPG [GCUV UVTCKP VJCV KPENWFGU VJG GPVKTG IGPGVKE DCEMITQWPFU QH VJG VYQ RCTGPVCN [GCUVU. 6TCPUHQTOCVKQP QHHGTU VJG RQUUKDKNKV[ VQ RTGEKUGN[ EJCPIG URGEKHKE EJCTCEVGTKUVKEU QH CP[ GZKUVKPI YKPG [GCUVU. #P GZKUVKPI RTQRGTV[ ECP DG OQFKHKGF, C PGY EJCTCEVGTKUVKE ECP DG KPVTQFWEGF YKVJQWV CFXGTUGN[ CHHGEVKPI QVJGT FGUKTCDNG RTQRGTVKGU, QT CP WPYCPVGF VTCKV ECP DG GNKOKPCVGF.
+P EQPVTCUV, IGPGVKE GPIKPGGTKPI OCMGU KV RQUUKDNG VQ EJCPIG VJG EJCTCEVGTKUVKEU QH YKPG [GCUVU YKVJ UWTIKECN RTGEKUKQP. 6JG VTCPUHQTOCVKQP QH S. cerevisiae EGNNU D[ HQTGKIP &ò# YCU HKTUV FGUETKDGF KP 1üøõ ($GIIU 1üøõ; *KPPGP GV CN. 1üøõ); UKPEG VJGP C NCTIG PWODGT QH IGPG ENQPKPI CPF VTCPUHQTOCVKQP UVTCVGIKGU, RNCUOKF XGEVQTU, UGNGEVCDNG OCTMGTU, RTQOQVGTU CPF NGCFGT UGSWGPEGU JCXG DGGP FGXGNQRGF HQT VJG GZRTGUUKQP QH JGVGTQNQIQWU IGPGU CPF VJG UGETGVKQP QH VJG RTQVGKPU OGCPV VQ HWPEVKQP KP VJG GZVTCEGNNWNCT EWNVWTG OGFKWO (6CDNG ÿ). +P RTCEVKEG, ENCUUKE DTGGFKPI CPF IGPGVKE GPIKPGGTKPI OGVJQFU CTG QHVGP WUGF VQIGVJGT VQ KORTQXG YKPG [GCUVU. #PF UKIPKHKECPVN[, XCTKCPVU, OWVCPVU, J[DTKFU CPF E[VQFWEVCPVU CTG PQV KPENWFGF KP VJG UVCVWVQT[ FGHKPKVKQP QH þ/ýU. 6JG[ CTG VJGTGHQTG PQV UWDLGEV VQ VJG UVCVWVQT[ TGIWNCVKQPU VJCV RGTVCKP VQ þ/ýU CPF CTG PQV VTGCVGF YKVJ VJG UCOG NGXGN QH RWDNKE UWURKEKQP CU CTG YKPG [GCUVU VJCV JCXG DGGP VTCPUHQTOGF YKVJ HQTGKIP &ò# (UGG 5GEVKQP ó).
10õ
+UCM 5. 2TGVQTKWU
6CDNG 2. /CKP UVCIGU KP JGVGTQNQIQWU IGPG GZRTGUUKQP KP YKPG [GCUV EGNNU 5VCIG
$TKGH FGUETKRVKQP CPF QRVKQPU CV GXGT[ UVCIG
þGPG KUQNCVKQP
# UVTWEVWTCN IGPG (QT, OQTG NKMGN[, EQORNGOGPVCT[ &ò#) GPEQFKPI VJG RTQVGKP QH KPVGTGUV KU KUQNCVGF HTQO TGUVTKEVKQP HTCIOGPVU QH &ò# HTQO C FQPQT QTICPKUO. &0# HTCIOGPVU EQPVCKPKPI VCTIGV IGPG - IGPQOKE HTCIOGPV - E&ò# HTCIOGPV - 2%4 HTCIOGPV - %JGOKECNN[ U[PVJGUKUGF HTCIOGPV
%NQPKPI (XGEVQTKUCVKQP)
6JG KP XKVTQ-OCPKRWNCVGF JGVGTQNQIQWU IGPG KU TGEQODKPGF KPVQ CP CRRTQRTKCVG GZRTGUUKQP XGEVQT. -
-
6TCPUHQTOCVKQP
5GNGEVKQP
©ZRTGUUKQP
+PVGITCVKPI RNCUOKFU - ð+R ðGCUV +PVGITCVKXG RNCUOKFU #WVQPQOQWUN[ TGRNKECVKPI RNCUOKFU - ð4R ðGCUV 4GRNKECVKPI RNCUOKFU - ð©R ðGCUV ©RKUQOCN RNCUOKFU - ð%R ðGCUV %GPVTQOGTKE RNCUOKFU 5RGEKCNKUGF RNCUOKFU - ðöR ðGCUV öKPGCT RNCUOKFU - ð#% ðGCUV #TVKHKEKCN %JTQOQUQOGU - ðïR ðGCUV ©ZRTGUUKQP RNCUOKFU - ð&R ðGCUV &KUKPVGITCVKQP RNCUOKFU
6JG XGEVQT KU KPVTQFWEGF KPVQ [GCUV EGNNU QT URJGTQRNCUVU D[ CWVQPQOQWU QT KPVGITCVKXG VTCPUHQTOCVKQP. 5RJGTQRNCUVU: EGNN YCNNU CTG FKIGUVGF YKVJ N[VKE GP\[OGU, 2©þ CFFGF, DTKGH JGCV UJQEM CPF TGIGPGTCVKQP. %JGOKECN: EGNNU CTG RGTOGCDKNKUGF YKVJ NKVJKWO UCNVU (G.I., CEGVCVG), &ò# CPF 2©þ CFFGF CPF EGNNU DTKGHN[ JGCV UJQEMGF. ©NGEVTKECN: EGNNU CPF &ò# CTG GNGEVTKECNN[ RWNUGF KP CP GNGEVTQRQTCVKQP FGXKEG $KQNKUVKE: &ò#-EQCVGF VWPIUVGP OKETQRTQLGEVKNG RCTVKENGU CTG RTQRGNNGF KP C ÂIGPG IWP• CV JKIJ URGGF KPVQ VJG EGNN. 5GNGEVCDNG IGPGVKE OCTMGTU, ECTTKGF D[ RNCUOKF XGEVQTU, CTG WUGF VQ KFGPVKH[ VTCPUHQTOCPVU (G.I., D[ EQORNGOGPVCVKQP QH C TGEGUUKXG OWVCVKQP KP VJG JQUV UVTCKP). ©ORKTKECN UGNGEVKQP (CHVGT OWVCIGPGUKU) OC[ DG GORNQ[GF HQT UWRGTUGETGVQT UVTCKPU. ÿGEGUUKXG OCTMGTU: HIS3, LEU2, LYS2, TRP1, URA3, ADE2, GVE. &QOKPCPV OCTMGTU: 4GUKUVCPEG VQ EQRRGT (CUP1), IGPGVKEKP þû1õ, UWRJQOGVWTQP (SMR1), J[ITQO[EKP, OGVJQVTGZCVG, EJNQTCOHGPKEQN, FKWTQP, ECPCXCPKP, \GQEKP, GVE. *GVGTQNQIQWU IGPG GZRTGUUKQP KPXQNXGU: VTCPUETKRVKQP WUKPI RTQOQVGT UGSWGPEGU HWUGF WRUVTGCO HTQO VJG UVTWEVWTCN IGPG; VGTOKPCVKQP QH VTCPUETKRVKQP WUKPI VGTOKPCVQT UGSWGPEGU HWUGF FQYPUVTGCO HTQO VJG UVTWEVWTCN IGPG; O4ò# VTCPURQTV HTQO PWENGQRNCUO VQ E[VQRNCUO; KPKVKCVKQP QH VTCPUNCVKQP CPF GNQPICVKQP QH VJG RQN[RGRVKFG EJCKP. %QPUVKVWVKXG RTQOQVGTU: ADH1, END1, GAP491, PGK1, PYK1, TPI1, GVE. ÿGIWNCVGF RTQOQVGTU: ADH2, CUP1, GAL1, GAL7, GAL10, HSP30 MEL1, MET25, MF½1, PHO5, GVE
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
10ü
6CDNG 2. (EQPVKPWGF) /QFKHKECVKQP 5KIPCNKPIñ 5GETGVKQP
*GVGTQNQIQWU RTQVGKPU CTG UWDLGEVGF VQ RQUV-VTCPUNCVKQPCN EJGOKECN OQFKHKECVKQP (G.I., IN[EQU[NCVKQP, CEGV[NCVKQP, GVE.) 5RGEKHKE UGETGVKQP UKIPCN RGRVKFGU CTG TGOQXGF HQT UGETGVQT[ RTQVGKPU, QT RTQVGKPU OC[ DG CNNQYGF VQ CEEWOWNCVG KP EGNNU QT URGEKHKE QTICPGNNGU. %GTVCKP RTQVGKPU, KH FGUKTGF, CTG GZRQTVGF HTQO VJG EGNN KPVQ VJG UWTTQWPFKPI OGFKWO. *QOQNQIQWU UGETGVKQP UKIPCNU: MEL1, MF½1, PHO5, STA2, SUC2, GVE. *GVGTQNQIQWU UGETGVKQP UKIPCNU: AMY1, BLA, IFN, GLU1, GVE.
4.4 5VTCVGIKGU CPF VCTIGVU HQT VJG KORTQXGOGPV QH YKPG [GCUVU +ORTQXGF YKPG [GCUV UVCTVGT EWNVWTG UVTCKPU CTG FGXGNQRGF VQ GPJCPEG EQUVGHHGEVKXGPGUU, GHHKEKGPE[ CPF UWUVCKPCDKNKV[ KP VJG RTQFWEVKQP QH JKIJ-SWCNKV[ YKPG. +ORTQXGOGPVU TGN[ JGCXKN[ QP DGVVGT WPFGTUVCPFKPI QH OGVCDQNKE HNWZGU, UKIPCNNKPI RCVJYC[U, IGPG TGIWNCVKQP CPF GZRTGUUKQP, CU YGNN CU QP RTQVGKP U[PVJGUKU CPF UGETGVKQP KP S. cerevisiae. 9KPG RTQFWEGTU CPF EQPUWOGTU WNVKOCVGN[ FKEVCVG VJG FKTGEVKQP QH VCTIGVGF UVTCKP FGXGNQROGPV, YJKEJ KP VWTP FGRGPFU WRQP CP WPFGTUVCPFKPI QH VJG EQORNGZ OGEJCPKUOU D[ YJKEJ [GCUV EGNNU CFCRV VQ VJG EQPVKPWQWUN[ EJCPIKPI GPXKTQPOGPV FWTKPI YKPGOCMKPI. 5QOG QH VJGUG OGEJCPKUOU CTG EQORNGZ CPF FKHHKEWNV VQ FGHKPG IGPGVKECNN[ YKVJQWV C DGVVGT WPFGTUVCPFKPI QH VJG DKQEJGOKUVT[, RJ[UKQNQI[ CPF IGPGVKEU KPXQNXGF. ðGCUV UVTCKP FGXGNQROGPV V[RKECNN[ IQGU VJTQWIJ CP KPKVKCN analysis phase, FWTKPI YJKEJ VJG HWPFCOGPVCNU QH VJG TGNCVGF CURGEVU CTG UVWFKGF VJQTQWIJN[ KP C NCDQTCVQT[ UVTCKP QH S. cerevisiae, HQNNQYGF D[ C synthesis phase, KP YJKEJ VJG CEVWCN EQPUVTWEVKQP QH VJG VCKNQTGF YKPG [GCUV UVTCKP QEEWTU (·UVGTICCTF GV CN. ÿ000). 6JKU EQPUGTXCVKXG CRRTQCEJ JCU TGUWNVGF KP UNQY DWV UVGCF[ RTQITGUU QXGT TGEGPV [GCTU KP VJG FGXGNQROGPV QH KORTQXGF YKPG [GCUVU (2TGVQTKWU CPF $CWGT ÿ00ÿ). 6JG HQNNQYKPI UGEVKQP FKUEWUUGU VJG RTQITGUU OCFG KP FGXGNQRKPI YKPG [GCUVU VJCV KORTQXG HGTOGPVCVKQP, RTQEGUUKPI CPF DKQRTGUGTXCVKQP, CPF KPETGCUG VJG YJQNGUQOGPGUU CPF UGPUQT[ SWCNKV[ QH YKPG. 6CDNG 3 UWOOCTKUGU VJG FKUEWUUKQP. 6CDNG 3. &GUKTCDNG EJCTCEVGTKUVKEU QH YKPG [GCUVU CPF VCTIGVU HQT IGPGVKE KORTQXGOGPV (CFCRVGF HTQO 2TGVQTKWU CPF $CWGT ÿ00ÿ) &GUKTCDNG RTQRGTVKGU +ORTQXGF HGTOGPVCVKQP RGTHQTOCPEG
+ORTQXGF IGPGTCN TGUKNKGPEG CPF UVTGUU VQNGTCPEG
(QEWU CTGCU
©ZCORNGU QH RQVGPVKCN VCTIGV IGPGU
5VTGUU TGURQPUG, UVGTQN, IN[EQIGP CPF VTGJCNQUG CEEWOWNCVKQP
/QFKHKECVKQP QH IN[EQIGP QT VTGJCNQUG OGVCDQNKUO =HQT GZCORNG CEVKPI QP GSY1 CPF GSY2 (IN[EQIGP U[PVJCUG), TPS1 (VTGJCNQUG-ù-RJQURJCVG U[PVJCUG), TPS2 (VTGJCNQUG-ù-RJQURJCVG RJQURJCVCUG)?
110
+UCM 5. 2TGVQTKWU
6CDNG 3. þEQPVKPWGFý +ORTQXGF GHHKEKGPE[ QH UWICT WVKNKUCVKQP
*GZQUG VTCPURQTVGTU, JGZQUG MKPCUGU
+ORTQXGF GHHKEKGPE[ QH PKVTQIGP CUUKOKNCVKQP
+ORTQXGF WVKNKUCVKQP QH NGUU GHHKEKGPV ò-UQWTEGU
+ORTQXGF GVJCPQN VQNGTCPEG +PETGCUGF VQNGTCPEG VQ CPVKOKETQDKCN EQORQWPFU
5VGTQN HQTOCVKQP, OGODTCPG #62CUG CEVKXKV[ 4GUKUVCPEG VQ MKNNGT VQZKPU, UWNRJWT FKQZKFG, CITQEJGOKECNU %GNN UWTHCEG RTQVGKPU
4GFWEGF HQCO HQTOCVKQP
+ORTQXGF RTQEGUUKPI GHHKEKGPE[ +ORTQXGF RTQVGKP ENCTKHKECVKQP +ORTQXGF RQN[UCEEJCTKFG ENCTKHKECVKQP
2TQVGCUGU
%QPVTQNNGF EGNN UGFKOGPVCVKQP CPF HNQEEWNCVKQP
÷NQEEWNKPU
%QPVTQNNGF EGNN HNQVCVKQP CPF HNQT HQTOCVKQP
%GNN YCNN J[FTQRJQDKE RTQVGKPU
+ORTQXGF YKPG YJQNGUQOGPGUU
+PETGCUGF RTQFWEVKQP QH TGUXGTCVTQN 4GFWEGF HQTOCVKQP QH GVJ[N ECTDCOCVG 4GFWEGF HQTOCVKQP QH DKQIGPKE COKPGU +ORTQXGF DKQRTGUGTXCVKXG ECRCDKNKVKGU
þNWECPCUGU, RGEVKPCUGU, Z[NCPCUGU, CTCDKPHWTCPQUKFCUGU
5VKNDGPG U[PVJGUKU #OKPQ CEKF OGVCDQNKUO, WTGC HQTOCVKQP $CEVGTKQN[VKE GP\[OGU, DCEVGTKQEKPU #PVKOKETQDKCN GP\[OGU CPF RGRVKFGU; HQTOCVKQP QH UWNRJKVGU
ýXGTGZRTGUUKQP CPF OQFKHKECVKQP QH HXT1HXT18, SNF3, FSY1 CPF WUG QH JGVGTQNQIQWU VTCPURQTVGTU CPF MKPCUGU 2TQNKPG ECVCDQNKUO =PUT1 (RTQNKPG QZKFCUG) CPF PUT2 (R[TTQNKPG-ó-ECTDQZ[NCVG FGJ[FTQIGPCUG)? CPF WUG QH JGVGTQNQIQWU ECVCDQNKE IGPGU /QFKHKECVKQP QH VJG GZRTGUUKQP QH PMA1 CPF PMA2 (#62CUG), UVGTQN CPCDQNKE IGPGU +PENWUKQP QH KIL2 (\[OQEKP CPF KOOWPKV[ HCEVQT), QXGTGZRTGUUKQP QH CUP1 (EQRRGT EJGNCVKP) &GNGVKQP QH FRO1 CPF FRO2 (HTQVJ RTQVGKPU)
ýXGTGZRTGUUKQP QH PEP4 (RTQVGCUG #) CPF UGETGVKQP QH QVJGT RTQVGCUGU ýXGTGZRTGUUKQP QH END1 (GPFQINWECPCUG), EXG1 (GZQINWECPCUG), CEL1 (EGNNQFGZVTKPCUG), BGL1 (¾-INWEQUKFCUG, EGNNQDKCUG), PEL5 (RGEVCVG N[CUG) CPF PEH1 (RQN[ICNCEVWTQPCUG), XYN1-5 (Z[NCPCUGU), ABF2 (CTCDKPQHWTCPQUKFCUG) öCVG GZRTGUUKQP QH HNQEEWNCVKQP IGPGU (FLO1, FLO5, MUC1/FLO11) WPFGT EQPVTQN QH RTQOQVGTU (*5230) KORCTVKPI FGUKTGF GZRTGUUKQP öCVG GZRTGUUKQP QH MUC1/FLO11 WPFGT EQPVTQN QH RTQOQVGTU (HSP30) KORCTVKPI FGUKTGF GZRTGUUKQP RCVVGTP
©ZRTGUUKQP QH 4CL9/216 (EQ-GP\[OG # NKICUG), VST1 (UVKNDGPG U[PVJCUG) &GNGVKQP QH CAR1 (CTIKPCUG) QT GZRTGUUKQP QH URE1 (WTGCUG) ©ZRTGUUKQP QH HEL1 (JGP GII YJKVG N[UQ\[OG), PED1 (RGFKQEKP), LCA1 (NGWEQEKP) CPF QVJGT DCEVGTKQEKPU ©ZRTGUUKQP QH HEL1 (JGP GII YJKVG N[UQ\[OG), CTS1 (EJKVKPCUG), EXG1 (GZQINWECPCUG) CPF QVJGT CPVKOKETQDKCN GP\[OGU; GZRTGUUKQP QH PED1 (RGFKQEKP), LCA1 (NGWEQEKP) CPF QVJGT JGVGTQNQIQWU DCEVGTKQEKP CPF \[OQEKP IGPGU; QXGTGZRTGUUKQP QH MET14 (CFGPQU[NRJQURJQUWNRJCVG MKPCUG) CPF MET16 (RJQURJQ CFGPQU[NRJQURJQUWNRJCVG TGFWEVCUG), CPF FGNGVKQP QH MET10 (UWNRJKVG TGFWEVCUG)
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
111
6CDNG 3. þEQPVKPWGFý &GETGCUGF NGXGNU QH CNEQJQN
%CTDQP HNWZ, IN[EGTQN OGVCDQNKUO CPF INWEQUG QZKFCVKQP
ýXGTGZRTGUUKQP QH GPD1 CPF GPD2 (IN[EGTQN-3-RJQURJCVG FGJ[FTQIGPCUG), OQFKHKECVKQP QH FPS1 (IN[EGTQN VTCPURQTV HCEKNKVCVQT), GZRTGUUKQP QH GOX1 (INWEQUG QZKFCUG)
©PJCPEGF NKDGTCVKQP QH ITCRG VGTRGPQKFU
þN[EQUKFCUGU, INWECPCUGU, CTCDKPQHWTCPQUKFCUGU
©PJCPEGF RTQFWEVKQP QH FGUKTCDNG XQNCVKNG GUVGTU
©UVGTCUGU
ýRVKOKUGF HWUGN QKN RTQFWEVKQP ©PJCPEGF IN[EGTQN RTQFWEVKQP
#OKPQ CEKF OGVCDQNKUO
ýXGTGZRTGUUKQP QH END1 (GPFQINWECPCUG), EXG1 (GZQINWECPCUG), CEL1 (EGNNQFGZVTKPCUG), BGL1 (¾-INWEQUKFCUG, EGNNQDKCUG), PEL5 (RGEVCVG N[CUG) CPF PEH1 (RQN[ICNCEVWTQPCUG), ABF2 (CTCDKPQHWTCPQUKFCUG) /QFKHKGF GZRTGUUKQP QH ATF1 (CNEQJQN CEGV[N VTCPUHGTCUG) CPF QVJGT CNEQJQN VTCPUHGTCUGU, IAH1 (GUVGTCUG) CPF QVJGT GUVGTCUGU &GNGVKQP QH VJG ILE, LEU CPF VAL IGPGU
$KQ-CFLWUVOGPV QH YKPG CEKFKV[
/CNQGVJCPQNKE CPF OCNQNCEVKE HGTOGPVCVKQP, NCEVKE CEKF RTQFWEVKQP
ýRVKOKUCVKQP QH RJGPQNKEU
2JGPQNKE CEKF OGVCDQNKUO
4GFWEGF UWNRJKVG CPF UWNRJKFG RTQFWEVKQP
5WNRJWT OGVCDQNKUO, J[FTQIGP UWNRJKFG HQTOCVKQP
+ORTQXGF YKPG HNCXQWT CPF QVJGT UGPUQT[ SWCNKVKGU
þN[EGTQN OGVCDQNKUO
ýXGTGZRTGUUKQP QH GPD1 CPF GPD2 (IN[EGTQN-3-RJQURJCVG FGJ[FTQIGPCUG), FPS1 (IN[EGTQN VTCPURQTV HCEKNKVCVQT), CPF FGNGVKQP QH ALD6 ©ZRTGUUKQP QH MAE1 (OCNCVG RGTOGCUG), VQIGVJGT YKVJ MAE2 (OCNKE GP\[OG) QT mleS (OCNQNCEVKE GP\[OG), QT LDH1 (NCEVKEQFGJ[FTQIGPCUG) /QFKHKGF GZRTGUUKQP QH PAD1 (RJGP[N CET[NKE CEKF FGECTDQZ[NCUG), pdc (ÐEQWOCTKE CEKF FGECTDQZ[NCUG), padc (RJGPQNKE CEKF FGECTDQZ[NCUG) &GNGVKQP QH MET14 (CFGPQU[NRJQURJQUWNRJCVG MKPCUG) CPF MRX1 (OGVJKQPKPG UWNRJQZKFG TGFWEVCUG)
4.4.1 +ORTQXGOGPV QH HGTOGPVCVKQP RGTHQTOCPEG 6JG RTGFKEVCDKNKV[ QH VJG HGTOGPVCVKQP QH ITCRG OWUV CPF VJG SWCNKV[ QH VJG YKPG FGRGPF FKTGEVN[ QP VJG ECRCDKNKV[ QH VJG YKPG [GCUV VQ TCRKFN[ GUVCDNKUJ PWOGTKECN CPF OGVCDQNKE FQOKPCPEG KP VJG GCTN[ RJCUG QH VJG HGTOGPVCVKQP, TGSWKTGF HQT CP GXGP CPF GHHKEKGPV HGTOGPVCVKQP YKVJ C FGUKTCDNG TGUKFWCN UWICT NGXGN (*GPUEJMG 1üüø). 6JG OQUV KORQTVCPV VCTIGVU HQT VJG KORTQXGOGPV QH HGTOGPVCVKQP RGTHQTOCPEG KPENWFG VJG HQNNQYKPI: KPETGCUGF TGUKNKGPEG CPF UVTGUU TGUKUVCPEG QH CEVKXG FTKGF [GCUV EGNNU; KORTQXGF ITCRG UWICT CPF PKVTQIGP WRVCMG CPF CUUKOKNCVKQP; GPJCPEGF TGUKUVCPEG VQ GVJCPQN CPF QVJGT OKETQDKCN OGVCDQNKVGU CPF VQZKPU; TGUKUVCPEG VQ UWNRJKVG, JGCX[ OGVCNU CPF CITQEJGOKECN TGUKFWGU; CPF TGFWEGF HQCO HQTOCVKQP (2TGVQTKWU ÿ000).
11ÿ
+UCM 5. 2TGVQTKWU
4.4.1.1 Improved resilience and stress tolerance 6JG KPCDKNKV[ QH YKPG [GCUV UVTCKPU VQ TGURQPF CPF CFCRV VQ WPHCXQWTCDNG, UVTGUUHWN EQPFKVKQPU OC[ NGCF VQ UVWEM QT UNWIIKUJ HGTOGPVCVKQPU CPF VJG RTQFWEVKQP QH QHHHNCXQWTU ($CWGT CPF 2TGVQTKWU ÿ000). 6JG KPJGTGPV TGUKNKGPEG, UVTGUU VQNGTCPEG, HKVPGUU CPF XKIQWT QH URGEKHKE UVTCKPU EQTTGNCVGU KPXGTUGN[ YKVJ VJG HTGSWGPE[ QH UVWEM QT UNWIIKUJ HGTOGPVCVKQPU (+XQTTC GV CN. 1üüü). 6JGUG CVVTKDWVGU TGNCVG VQ DQVJ EGNN XKCDKNKV[ (VJG TGNCVKXG RTQRQTVKQP QH NKXKPI EGNNU YKVJKP C RQRWNCVKQP) CPF XKVCNKV[ (VJG OGCUWTG QH OGVCDQNKE CEVKXKV[) FWTKPI VJG RTQEGUU QH [GCUV OCPWHCEVWTKPI CPF YKPG HGTOGPVCVKQP (9CNMGT 1üüõ). #U KU VJG ECUG HQT DCMGT•U [GCUV (UGG %JCRVGT 3), VJG EJQKEG QH C UVTGUU-VQNGTCPV UVTCKP CPF VJG URGEKHKE RTGRCTCVKQP RTQEGFWTG WUGF VQ RTQFWEG C EQOOGTEKCN CEVKXG FTKGF YKPG [GCUV UVCTVGT EWNVWTG ECP CHHGEV VJG FGITGG QH XKCDKNKV[ CPF XKVCNKV[, CU YGNN CU VJG UWDUGSWGPV HGTOGPVCVKQP RGTHQTOCPEG QH VJG EWNVWTGU ($CWGT CPF 2TGVQTKWU ÿ000; þCIKCPQ GV CN. ÿ00ÿ). +V KU IGPGTCNN[ TGEQOOGPFGF VJCV VJG OCPWHCEVWTGTU QH VJGUG UVCTVGT EWNVWTGU EWNVKXCVG VJGKT [GCUV VQ OCZKOKUG COQWPVU QH UVGTQNU (G.I., GTIQUVGTQN) CPF UVQTCIG ECTDQJ[FTCVGU (G.I., VTGJCNQUG CPF IN[EQIGP) KP VJG [GCUV EGNNU. 5VGTQNU CPF VTGJCNQUG RNC[ CP GUUGPVKCN TQNG KP VJG OGCUWTG QH [GCUV EGNNU• TGUKUVCPEG VQ FT[KPI CPF UWDUGSWGPV TGJ[FTCVKQP D[ UVCDKNKUKPI VJG [GCUV'U OGODTCPG UVTWEVWTG, YJGTGCU IN[EQIGP RTQXKFGU C TGCFKN[ OQDKNKUCDNG ECTDQP CPF GPGTI[ UQWTEG FWTKPI VJG CFCRVCVKQP RJCUG WRQP TGCEVKXCVKQP (&GITo, 1üü3; 9CNMGT 1üüõ). 6JWU VJGTG KU C UVTQPI KPEGPVKXG VQ FGXGNQR YKPG [GCUV UVTCKPU YKVJ C UWRGTKQT CDKNKV[ VQ CEEWOWNCVG VJG UVGTQNU, VTGJCNQUG CPF IN[EQIGP. 6JG QXGTGZRTGUUKQP QH UQOG IGPGU UWEJ CU SUT1 CPF SUT2 KPXQNXGF KP VJG CEEWOWNCVKQP QH UVGTQNU JCU DGGP UJQYP VQ RTQOQVG VJG WRVCMG QH UVGTQN HTQO VJG OGFKWO WPFGT HGTOGPVCVKXG EQPFKVKQPU. *QYGXGT, CVVGORVU VQ KPETGCUG UVTGUU TGUKUVCPEG VJTQWIJ VJG QXGTGZRTGUUKQP QH VJG TPS1-GPEQFGF VTGJCNQUG-ù-RJQURJCVG U[PVJGVCUG, TPS2-GPEQFGF VTGJCNQUG-ù-RJQURJCVG RJQURJCVCUG, CPF VJG GSY1- CPF GSY2-GPEQFGF IN[EQIGP U[PVJCUG JCXG PQV [GV GUVCDNKUJGF C EQPXKPEKPI NKPGCT NKPM DGVYGGP VTGJCNQUGñIN[EQIGP EQPVGPV CPF UVTGUU TGUKUVCPEG (6JGXGNGKP 1üüù; 6JGXGNGKP CPF &G 9KPFG 1üüü; 2TGVQTKWU CPF $CWGT ÿ00ÿ). *QYGXGT, C [GCUV UVTCKP FGNGVGF HQT VJG ATH1 VTGJCNCUG IGPG EQPVCKPGF JKIJGT NGXGNU QH VTGJCNQUG CPF UJQYGF KORTQXGF UWTXKXCN CHVGT FGJ[FTCVKQP, HTGG\KPI CPF GVJCPQN UJQEM (îKO GV CN. 1üüù; 5JKOC GV CN. 1üüü). 6JG QXGTGZRTGUUKQP QH VYQ CSWCRQTKP IGPGU (AQY1 CPF AQY2) YCU TGEGPVN[ UJQYP VQ UKIPKHKECPVN[ GPJCPEG VJG HTGG\G TGUKUVCPEG QH DCMGT•U [GCUVU (8CP &KLEM GV CN. ÿ000; 6CPIJG GV CN. ÿ000, ÿ00ÿ; 6GWPKUUGP GV CN. ÿ00ÿ). 6JGUG HKPFKPIU CTG UKIPKHKECPV UVGRU CNQPI VJG RCVJ VQYCTF GPJCPEKPI UVTGUU TGUKUVCPEG KP YKPG [GCUVU. 4.4.1.2 Improved efficiency of sugar and nitrogen utilisation 6JG CUUKOKNCDNG ECTDQP CPF PKVTQIGP EQPVGPV QH ITCRG OWUV KU FGRGPFGPV WRQP VJG ITCRG EWNVKXCT CPF TQQVUVQEM, CU YGNN CU QP UGXGTCN CURGEVU QH XKPG[CTF OCPCIGOGPV CPF YKPGOCMKPI RTCEVKEG (G.I., UQKN V[RG, PKVTQIGP HGTVKNKUCVKQP, XKPG YCVGT UVCVWU, ITCRGXKPG JGCNVJ, DGTT[ OCVWTCVKQP, TKRGPGUU QH ITCRGU, EJGOKECN CFLWUVOGPVU, CFFKVKXGU KP VJG ITCRG OWUV) (*GPUEJMG CPF íKTCPGM 1üü3; 2TGVQTKWU ÿ000,
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
113
ÿ003). þTCRG UWICTU (OCKPN[ INWEQUG CPF HTWEVQUG) CTG WUWCNN[ RTGUGPV KP GZEGUU QH YJCV KU PGGFGF HQT OCZKOCN [GCUV ITQYVJ. +P EQPVTCUV, VJG CUUKOKNCDNG PKVTQIGP EQPVGPV KP ITCRG OWUV (OCKPN[ COKPQ CEKFU CPF COOQPKWO) KU QHVGP NKOKVGF, OQUVN[ DGECWUG [GCUV EGNNU ECPPQV VCMG WR RTQNKPG (YJKEJ, VQIGVJGT YKVJ CTIKPKPG, CEEQWPVU HQT 30-ùóì QH ITCRG COKPQ CEKFU) WPFGT CPCGTQDKE EQPFKVKQPU FWTKPI YKPG HGTOGPVCVKQP; VJG[ CNUQ ECPPQV J[FTQN[UG ITCRG RTQVGKPU VQ UWRRNGOGPV PKVTQIGP-FGHKEKGPV OWUVU. 2QQT HGTOGPVCVKXG RGTHQTOCPEG JCRRGPU OQUV EQOOQPN[ YJGP VJGTG KU CP KODCNCPEG KP VJG JKIJ NGXGNU QH ECTDQP CPF NQY NGXGNU QH PKVTQIGP KP VJG ITCRG OWUV. 5NWIIKUJ QT UVWEM HGTOGPVCVKQPU QEEWT YJGP PKVTQIGP FGRNGVKQP NKOKVU VJG TCVG QH UWICT VTCPURQTV ($KUUQP 1üüü). òKVTQIGP NKOKVCVKQP FWTKPI YKPG HGTOGPVCVKQP NGCFU VQ VJG KPVGTPCNKUCVKQP CPF FGITCFCVKQP QH VJG JGZQUG VTCPURQTVGTU, VJGTGD[ KTTGXGTUKDN[ CTTGUVKPI JGZQUG VTCPURQTV ($QNGU CPF *QNNGPDGTI 1üüø; $KUUQP 1üüü; f\ECP CPF íQJPUQP 1üüü). 6JGTG CRRGCTU VQ DG CV NGCUV VJTGG YC[U VQ GPUWTG VJG GHHKEKGPV WVKNKUCVKQP QH ITCRG UWICT WPFGT EQPFKVKQPU QH PKVTQIGP NKOKVCVKQP: KPETGCUG VJG TCVG QH IN[EQN[VKE HNWZ D[ TGRNCEKPI CP[ PQP-HWPEVKQPCN OWVCPV CNNGNG QH IGPGU GPEQFKPI VJG MG[ IN[EQN[VKE GP\[OGU; GPJCPEG VJG GHHKEKGPE[ QH JGZQUG (GURGEKCNN[ HTWEVQUG) WRVCMG; CPF CNNGXKCVG VJG CUUKOKNCVKQP QH RTQNKPG CPF CTIKPKPG HTQO PKVTQIGP ECVCDQNKVG TGRTGUUKQP. 6JWU HCT, QPN[ NKOKVGF UWEEGUU JCU DGGP CEJKGXGF KP VJKU TGICTF. (2TGVQTKWU CPF $CWGT ÿ00ÿ). 6JG QXGTGZRTGUUKQP QH VJG IGPGU GPEQFKPI UGXGP GP\[OGU KPXQNXGF KP VJG NCVVGT UVCIGU QH IN[EQN[UKU JCU NGF VQ KPETGCUGF IN[EQN[VKE HNWZ, DWV QPN[ WPFGT EQPFKVKQPU QH KPETGCUGF #62 FGOCPF (5OKVU GV CN. ÿ000). +P VJG ECUG QH KPEQORNGVG HGTOGPVCVKQPU, VJG RTGHGTGPEG QH YKPG [GCUVU HQT INWEQUG QXGT HTWEVQUG ECP NGCF VQ GZEGUUKXG TGUKFWCN HTWEVQUG NGXGNU VJCV EQORTQOKUG VJG SWCNKV[ QH VJG YKPG. 5KPEG VJG TCVG QH CPCGTQDKE HGTOGPVCVKQP D[ [GCUV KU NKOKVGF RTKOCTKN[ D[ VJG TCVG QH UWICT WRVCMG (GURGEKCNN[ FWTKPI VJG HKPCN HGTOGPVCVKQP UVCIGU QH PKVTQIGP FGRNGVKQP EQWRNGF VQ PWVTKGPV NKOKVCVKQP), UGXGTCN NCDQTCVQTKGU HQEWU QP RJQURJQT[NCVKQP D[ VJG HXK1CPF HXK2-GPEQFGF JGZQMKPCUGU CPF VJG GLK1-GPEQFGF INWEQMKPCUG, CU YGNN CU QP JGZQUG UGPUQTU (GPEQFGF D[ SNF3 CPF RGT2) CPF JGZQUG VTCPURQTVGTU (GPEQFGF D[ HXT1-HXT18 CPF GAL2) QH S. cerevisiae (îTWEMGDGTI 1üüù; $QNGU CPF *QNNGPDGTI 1üüø). #V NGCUV HKXG QH VJGUG JGZQUG VTCPURQTVGTU (*ZV1-3, *ZVù-ø) QH S. cerevisiae CTG TGSWKTGF HQT C UWEEGUUHWN YKPG HGTOGPVCVKQP, RNC[KPI FKHHGTGPV TQNGU CV XCTKQWU UVCIGU KP VJG HGTOGPVCVKQP E[ENG (öW[VGP GV CN. ÿ00ÿ). 6JG NQY-CHHKPKV[ VTCPURQTVGTU (*ZV1 CPF *ZV3) RNC[ C RTGFQOKPCPV TQNG KP YKPG HGTOGPVCVKQP; *ZV3 RNC[U C OCLQT TQNG, YJKNG *ZV1•U TQNG KU TGUVTKEVGF VQ VJG DGIKPPKPI QH VJG YKPG HGTOGPVCVKQP (öW[VGP GV CN. ÿ00ÿ). 6JG JKIJ-CHHKPKV[ ECTTKGTU (*ZVÿ, *ZVù CPF *ZVø) CTG CNUQ TGSWKTGF CV FKHHGTGPV UVCIGU QH YKPG HGTOGPVCVKQP; *ZVÿ KU KPXQNXGF KP ITQYVJ KPKVKCVKQP, YJGTGCU *ZVù CPF *ZVø CTG TGSWKTGF CV VJG GPF QH CNEQJQNKE HGTOGPVCVKQP (öW[VGP GV CN. ÿ00ÿ). +V KU J[RQVJGUKUGF VJCV VJG QXGTGZRTGUUKQP QH VJG S. pastorianus FSY1-GPEQFGF HTWEVQUGñ*ë U[ORQTVGT, VQIGVJGT YKVJ VJG HXT1-3 CPF HXT6-GPEQFGF JGZQUG VTCPURQTVGTU CPF VJG HXK1-GPEQFGF JGZQMKPCUG (YKVJ VJG JKIJGUV CHHKPKV[ HQT HTWEVQUG, DWV UVKNN C UKIPKHKECPVN[ NQYGT CHHKPKV[ VJCP HQT INWEQUG), TGUWNVU KP KORTQXGF INWEQUG CPF HTWEVQUG WRVCMG FWTKPI YKPG HGTOGPVCVKQPU (2TGVQTKWU ÿ003). ÷WTVJGTOQTG, VJG FGNGVKQP QH VJG URE2-GPEQFGF TGRTGUUQT QH VJG PUT1-GPEQFGF RTQNKPG
11û
+UCM 5. 2TGVQTKWU
QZKFCUG CPF PUT2-GPEQFGF R[TTQNKPG-ó-ECTDQZ[NCVG FGJ[FTQIGPCUG TGRTGUGPVU VJG HKTUV UVGR VQYCTFU VJG FGXGNQROGPV QH YKPG [GCUVU VJCV ECP GHHKEKGPVN[ CUUKOKNCVG VJG CDWPFCPV UWRRN[ QH RTQNKPG CPF CTIKPKPG KP ITCRG LWKEG WPFGT HGTOGPVCVKXG EQPFKVKQPU (5CNOQP CPF $CTTG 1üüõ). +V KU ENGCT VJCV OQTG KPHQTOCVKQP KU TGSWKTGF CDQWV VJG EQORNGZ TGIWNCVKQP QH JGZQUG CPF PKVTQIGP WRVCMG CPF CUUKOKNCVKQP CU KV QEEWTU KP ITCRG LWKEG, GURGEKCNN[ KP VJG RTGUGPEG QH JKIJ UWICT NGXGNU FWTKPI VJG GCTN[ RJCUG QH HGTOGPVCVKQP CPF FWTKPI VJG HKPCN UVCIGU QH UWICT FGRNGVKQP EQWRNGF VQ PKVTQIGP NKOKVCVKQP CPF JKIJ NGXGNU QH GVJCPQN. 5WEJ KPHQTOCVKQP YKNN RTQOQVG PGY UVTCVGIKGU VQ KORTQXG VJG HGTOGPVCVKQP RGTHQTOCPEG QH YKPG [GCUV CPF VQ RTGXGPV UNWIIKUJ QT UVWEM HGTOGPVCVKQPU. 4.4.1.3 Increased tolerance to antimicrobial compounds 8CTKQWU [GCUV OGVCDQNKVGU (G.I., CNEQJQNU, CEGVKE CEKF CPF OGFKWO EJCKP NGPIVJ HCVV[ CEKFU) CPF QVJGT CPVKOKETQDKCN EQORQWPFU, UWEJ CU \[OQEKPU (MKNNGT VQZKPU), EJGOKECN RTGUGTXCVKXGU (GURGEKCNN[ UWNRJKVG) CPF CITQEJGOKECNU EQPVCKPKPI JGCX[ OGVCNU (G.I., EQRRGT), ECP KORGFG VJG HGTOGPVCVKQP RGTHQTOCPEG QH YKPG [GCUVU. S. cerevisiae UVTCKPU XCT[ YKFGN[ KP VJGKT CDKNKV[ VQ TGUKUV QT VQNGTCVG VJGUG EQORQWPFU. %QORCTGF VQ PQP-YKPG Saccharomyces UVTCKPU, EQOOGTEKCN YKPG [GCUV UVTCKPU EWNVKXCVGF WPFGT JKIJN[ CGTQDKE EQPFKVKQPU CPF KP NQY INWEQUG EQPEGPVTCVKQPU IGPGTCNN[ EQPVCKP JKIJGT NGXGNU QH VJG UQ-ECNNGF RTQVGEVKXG QT UWTXKXCN HCEVQTU, COQPI VJGO EGTVCKP WPUCVWTCVGF NQPI EJCKP HCVV[ CEKFU, UVGTQNU, KOOWPKV[ HCEVQTU CPF EJGNCVKPI RTQVGKPU. 6JG[ VJGTGD[ FKURNC[ C ITGCVGT RJ[UKQNQIKECN TGURQPUG VQ EJCNNGPIGU D[ VJGUG CPVKOKETQDKCN UWDUVCPEGU ($QWNVQP GV CN. 1üüù). 6JG IGPGVKEU QH GVJCPQN VQNGTCPEG CTG RQN[XCNGPV CPF VJG RJ[UKQNQIKECN DCUKU QH GVJCPQN VQZKEKV[ KU EQORNGZ CPF PQV YGNN WPFGTUVQQF. +V CRRGCTU VJCV GVJCPQN KORCEVU OCKPN[ QP VJG UVTWEVWTCN KPVGITKV[ CPF RGTOGCDKNKV[ QH VJG RNCUOC OGODTCPG CPF VJG OGODTCPGU QH VJG OKVQEJQPFTKC, PWENGWU CPF XCEWQNC, CU YGNN CU QP VJG GPFQRNCUOKE TGVKEWNWO CPF E[VQUQNKE J[FTQRJKNKE RTQVGKPU ($QWNVQP GV CN. 1üüù; 9CNMGT 1üüõ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ûó0, CNEQJQN FGJ[FTQIGPCUG CEVKXKV[ CPF GVJCPQN OGVCDQNKUO ($QWNVQP GV CN. 1üüù; *GPUEJMG 1üüø; 9CNMGT 1üüõ). 6JWU HCT, QYKPI VQ VJG EQORNGZKV[ QH VJGUG RTQVGEVKXG CFCRVCVKQPU VQ GVJCPQN, UWDUVCPVKCN KPETGCUGU KP VJG GVJCPQN VQNGTCPEG QH YKPG [GCUVU JCXG GNWFGF TGUGCTEJGTU. ÷QT VJG VKOG DGKPI, VJGTGHQTG, GVJCPQN VQNGTCPEG KP YKPG [GCUVU YKNN DG CF-
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
11ó
FTGUUGF D[ VJG OCPKRWNCVKQP QH VJG RTGXCKNKPI HGTOGPVCVKQP EQPFKVKQPU (fermentation engineering) CPF EGNN RJ[UKQNQIKECN CURGEVU (cell engineering) TCVJGT VJCP HQEWUUKPI UQNGN[ QP VJG OQFKHKECVKQP QH VJG [GCUV•U IGPQV[RG (genetic engineering) (9CNMGT 1üüõ). 6JG \[OQEKPU (MKNNGT VQZKPU GPEQFGF D[ GKVJGT PQP-KPHGEVKXG O[EQXKTWUGU QT [GCUV EJTQOQUQOCN IGPGU) RTQFWEGF D[ MKNNGT UVTCKPU (î1, îÿ, î3, îÿõ) QH S. cerevisiae CTG NGVJCN VQ UGPUKVKXG UVTCKPU QH VJG UCOG URGEKGU, YJKNG VJQUG RTQFWEGF D[ PQP-Saccharomyces MKNNGT URGEKGU (îû VQ î11) OC[ DG VQZKE VQ C YKFGT TCPIG QH YKPG [GCUV UVTCKPU CPF VQ QVJGT YKNF [GCUVU. îÿ CPF îÿõ, VJG VYQ S. cerevisiae MKNNGT VQZKPU VJCV HWPEVKQP CV YKPG R*, MKNN UGPUKVKXG YKPG [GCUVU XKC VYQ OGEJCPKUOU: VJG îÿ VQZKP CEVU CU CP KQPQRJQTG CHHGEVKPI OGODTCPG RGTOGCDKNKV[ CPF VJG NGCMCIG QH RTQVQPU, RQVCUUKWO ECVKQPU, #62 CPF COKPQ CEKFU, YJGTGCU VJG îÿõ VQZKP KPJKDKVU &ò# U[PVJGUKU (5EJOKVV GV CN. 1üüù). 7PFGT EGTVCKP EQPFKVKQPU VJCV HCXQWT VJG FGXGNQROGPV QH MKNNGT [GCUV EQPVCOKPCVKQP QH ITCRG LWKEG (G.I., KPGHHKEKGPV KPQEWNCVKQP YKVJ JKIJN[ UGPUKVKXG UVCTVGT EWNVWTGU KP NQY-PKVTQIGP OWUVU), RQVGPV \[OQEKFCN YKNF [GCUVU OC[ EQPVTKDWVG VQ UNWIIKUJ QT UVWEM HGTOGPVCVKQPU CPF RTQFWEG YKPGU YKVJ KPETGCUGF NGXGNU QH CEGVCNFGJ[FG, NCEVKE CEKF CPF CEGVKE CEKF CPF QVJGT WPFGUKTCDNG UGPUQT[ SWCNKVKGU (5JKOK\W 1üü3;8CFCU\ GV CN. ÿ00ÿ). 6JGTGHQTG, KP QTFGT VQ FGXGNQR DTQCF-URGEVTWO \[OQEKFCN TGUKUVCPEG KP YKPG [GCUVU, UQOG NCDQTCVQTKGU JCXG WUGF E[VQFWEVKQP CPF J[DTKFKUCVKQP OGVJQFU VQ VTCPUHGT \[OQEKP-GPEQFKPI O[EQXKTCN IGPQOGU HTQO MKNNGT [GCUVU KPVQ EQOOGTEKCN YKPG UVTCKPU. #VVGORVU CTG CNUQ DGKPI OCFG VQ ENQPG CPF KPVTQFWEG VJG VQZKP-KOOWPKV[ IGPGU HTQO PQP-Saccharomyces MKNNGT [GCUVU KPVQ YKPG [GCUVU. 6QNGTCPEG VQ EJGOKECN RTGUGTXCVKXGU UWEJ CU UWNRJKVG HQTOU VJG DCUKU QH VJG UGNGEVKXG KORNCPVCVKQP QH CEVKXG FTKGF YKPG [GCUV UVCTVGT EWNVWTGU KPVQ ITCRG OWUV (*GPUEJMG 1üüø). 5ýÿ FKHHWUGU KPVQ [GCUV EGNNU, FKUUQEKCVGU VQ 5ý3ÿ- CPF *5ý3 , NQYGTU VJG KPVTCEGNNWNCT R* CPF KPJKDKVU OGVCDQNKE CEVKXKV[ (9CNMGT 1üüõ). 6JQWIJ S. cerevisiae VQNGTCVGU OWEJ JKIJGT NGXGNU QH UWNRJKVG VJCP OQUV WPYCPVGF [GCUVU CPF DCEVGTKC, GZEGUUKXG 5ýÿ FQUCIGU OC[ ECWUG UNWIIKUJ QT UVWEM HGTOGPVCVKQPU ($QWNVQP GV CN. 1üüù). 9KPG [GCUVU FKHHGT EQPUKFGTCDN[ KP UWNRJKVG VQNGTCPEG. ýPEG VJG WPFGTN[KPI OGEJCPKUO QH 5ýÿ VQNGTCPEG CPF VJG IGPGVKE DCUKU HQT TGUKUVCPEG KP YKPG [GCUVU CTG DGVVGT WPFGTUVQQF, KV OC[ DG CFXCPVCIGQWU VQ GPIKPGGT YKPG [GCUV UVCTVGT UVTCKPU YKVJ GNGXCVGF 5ýÿ VQNGTCPEG. *QYGXGT, VJKU UJQWNF PQV TGRNCEG GHHQTVU VQ NQYGT VJG NGXGNU QH EJGOKECN RTGUGTXCVKXGU KP YKPG. 6JG KORTQRGT CRRNKECVKQP QH CITQEJGOKECNU, UWEJ CU CPVKHWPICN RGUVKEKFGU EQPVCKPKPI JGCX[ OGVCNU, ECP TGUWNV KP EQRRGT TGUKFWGU KP OWUVU, NGCFKPI VQ NCIIKPI HGTOGPVCVKQP CPF C FGVTKOGPVCN GHHGEV QP YKPG SWCNKV[ (6TQOR CPF &G îNGTM 1üõõ). %QRRGT VQZKEKV[ VQYCTFU YKPG [GCUV EGNNU KPXQNXGU VJG FKUTWRVKQP QH RNCUOC OGODTCPG KPVGITKV[ CPF QH VJG KPVTCEGNNWNCT KPVGTCEVKQP DGVYGGP EQRRGT CPF PWENGKE CEKFU CPF GP\[OGU (#XGT[ GV CN. 1üüù). 9JGP [GCUV EGNNU CTG GZRQUGF VQ RQVGPVKCNN[ NGVJCN NGXGNU QH EQRRGT, OGVCNNQVJGKP RTQVGKPU, UWEJ CU VJG CUP1-GPEQFGF EQRRGT EJGNCVKP, CTG U[PVJGUKUGF VQ UGSWGUVGT VJG OGVCN. 5KPEG VJG EQRRGT TGUKUVCPEG NGXGN QH C IKXGP [GCUV UVTCKP EQTTGNCVGU FKTGEVN[ YKVJ VJG CUP1 EQR[ PWODGT, VJKU IGPG
11ù
+UCM 5. 2TGVQTKWU
JCU DGGP KPVGITCVGF KPVQ VJG IGPQOGU QH UGXGTCN KPFWUVTKCN [GCUVU CV OWNVKRNG UKVGU (÷QIGN GV CN. 1üõ3; *GPFGTUQP GV CN. 1üõó). ýDXKQWUN[, UWEJ EQRRGT-TGUKUVCPV YKPG [GCUVU UJQWNF PQV DG CNNQYGF VQ GPEQWTCIG FKUTGURGEV HQT TGEQOOGPFGF HWPIKEKFG YKVJJQNFKPI RGTKQFU. 4.4.2 +ORTQXGOGPV QH RTQEGUUKPI GHHKEKGPE[ ÷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ñQT HKNVTCVKQP VQ TGOQXG EQNNQKFU, RCTVKCNN[ UQNWDNG JC\GHQTOKPI RTQVGKPU, HKNVGT-ENQIIKPI RQN[UCEEJCTKFGU, CU YGNN CU EQORNGZGU DGVYGGP RTQVGKPU CPF RJGPQNU CPF DGVYGGP RTQVGKPU CPF RQN[UCEEJCTKFGU (%CPCN-öNCWDGTGU 1üü3; %QNCITCPFG GV CN. 1üüû; $QWNVQP GV CN. 1üüù). 6JGUG CTG GZRGPUKXG CPF NCDQTKQWU RTCEVKEGU VJCV IGPGTCVG NCTIG XQNWOGU QH NGGU HQT FKURQUCN, VJGTGD[ ECWUKPI C NQUU QH YKPG CPF TGOQXKPI KORQTVCPV CTQOC CPF HNCXQWT EQORQWPFU HTQO VJG TGOCKPKPI YKPG. 4.4.2.1 Improved protein and polysaccharide clarification 2TQVGKP KPUVCDKNKV[ CPF VJG ITCFWCN FGXGNQROGPV QH RTQVGKP JC\GU KP YJKVG YKPG CTG ECWUGF OCKPN[ D[ VJG ITCRG•U RCVJQIGPGUKU-TGNCVGF (24) RTQVGKPU. 5QOG YKPGOCMGTU DGNKGXG VJCV VJGUG JC\G-HQTOKPI RTQVGKPU CTG FGITCFGF FWTKPI RTQNQPIGF UVQTCIG QH YKPG QP VJG [GCUV NGGU CHVGT VJG EQORNGVKQP QH VJG CNEQJQNKE HGTOGPVCVKQP, VJGTGD[ TGPFGTKPI VJG YKPG OQTG RTQVGKP UVCDNG ($QWNVQP GV CN. 1üüù; 4KDoTGCWþC[QP GV CN. ÿ000). 6JKU KU CVVTKDWVGF VQ VJG TGNGCUG QH VJG PEP4-GPEQFGF XCEWQNCT RTQVGCUG # HTQO VJG CWVQN[UKPI [GCUV EGNNU FWTKPI VJG sur lie RTQEGUU (2TGVQTKWU ÿ000). +P CP CVVGORV VQ OCMG VJG WUG QH DGPVQPKVG CPF EQOOGTEKCN GP\[OG RTGRCTCVKQPU WPPGEGUUCT[ HQT VJG TGOQXCN QH VJGUG JC\G-HQTOKPI RTQVGKPU, VJG GZRTGUUKQP NGXGNU QH VJG PEP4 IGPG YGTG KPETGCUGF, EQFKPI HQT VJG QPN[ S. cerevisiae RTQVGCUG CEVKXG CV VJG NQY R* QH YKPG. 6JG QXGTGZRTGUUKQP QH VJG PEP4 IGPG UGGOU VQ UCVWTCVG VJG XCEWQNCT VCTIGVKPI OCEJKPGT[, VJGTGD[ FKTGEVKPI VJG GZEGUU COQWPV QH RTQVGCUG # VQ VJG GZVGTPCN GPXKTQPOGPV (8CP 4GPUDWTI CPF 2TGVQTKWU ÿ000). *QYGXGT, VJG HGTOGPVCVKQP QH YJKVG YKPG YKVJ VJKU RTQVGQN[VKE [GCUV YCU WPCDNG VQ GHHGEVKXGN[ TGOQXG VJG JC\G-HQTOKPI RTQVGKPU CPF VQ GPUWTG RTQVGKP UVCDKNKV[ KP VJG HKPCN RTQFWEV. 6JKU KU OQUV NKMGN[ DGECWUG VJG URGEKHKE EQPHQTOCVKQP QH VJG ITCRG•U JC\GHQTOKPI 24 RTQVGKPU OCMGU VJGO KPJGTGPVN[ TGUKUVCPV VQ RTQVGQN[UKU. 5Q HCT, PQ RTQVGCUG JCU DGGP FGUETKDGF VJCV KU CDNG VQ QXGTEQOG VJKU TGECNEKVTCPEG QH VJG 24 RTQVGKPU, VJGTGD[ RWVVKPI VJG FGXGNQROGPV QH RTQVGQN[VKE YKPG [GCUVU QP JQNF HQT VJG VKOG DGKPI (2TGVQTKWU ÿ003).
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
11ø
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½-1,û-IN[EQUKFKE NKPMCIGU KP VJG RQN[ICNCEVWTQPCVG EJCKP (÷KI. óC). ÷KIWTG óD FGOQPUVTCVGU JQY INWECPU CTG J[FTQN[UGF D[ GPFQINWECPCUGU, GZQINWECPCUGU, EGNNQFGZVTKPCUGU CPF EGNNQDKCUGU (C OGODGT QH VJG ¾-INWEQUKFCUG HCOKN[) (2TGVQTKWU 1üüø). 6JG GP\[OCVKE FGITCFCVKQP QH Z[NCPU KU ECVCN[UGF D[ VJG U[PGTIKUVKE CEVKQPU QH GPFQ-¾-1,û-&-Z[NCPCUGU, ¾-&-Z[NQUKFCUGU CPF ½-ö-CTCDKPQHWTCPQUKFCUGU (÷KI. óE) (2TGVQTKWU 1üüø). 5QOG UVTCKPU QH S. cerevisiae YGTG TGRQTVGF VQ RTQFWEG RGEVKPGUVGTCUG, RQN[ICNCEVWTQPCUG CPF RGEVKP N[CUG (þCKPXQTU GV CN. 1üüû), YJKNG CNN UVTCKPU QH S. cerevisiae UJQY UQOG HQTO QH INWECPCUG CEVKXKV[ (2TGVQTKWU 1üüø; ö[PF GV CN. ÿ00ÿ). S. cerevisiae•U PGU1 IGPG (CNUQ MPQYP CU PGL1) GPEQFGU C RQN[ICNCEVWTQPCUG (þQIPKGU GV CN. 1üüü; þQIPKGU CPF $GNCTDK ÿ001; þQIPKGU GV CN. ÿ001). 6JG EXG1 (BGL1) IGPG GPEQFGU C RTQVGKP YJQUG FKHHGTGPVKCN IN[EQU[NCVKQP CEEQWPVU HQT VJG VYQ OCKP GZVTCEGNNWNCT GZQ-¾-1,3-INWECPCUGU (©ïþ+ CPF ©ïþ++), YJKNG EXG2 GPEQFGU C OKPQT GZQ-¾-1,3-INWECPCUG (©ïþ+++). BGL2 GPEQFGU C EGNN YCNNCUUQEKCVGF GPFQ-¾-1,3-INWECPCUG, YJKNG SSG1 (SPR1) EQFGU HQT C URQTWNCVKQPURGEKHKE GZQ-¾-1,3-INWECPCUG. 6JGUG GPFQIGPQWU RGEVKPQN[VKE CPF INWECPQN[VKE CEVKXKVKGU QH S. cerevisiae CTG PQV UWHHKEKGPV VQ GNKOKPCVG ENCTKHKECVKQP CPF HKNVTCVKQP RTQDNGOU. # YKFG XCTKGV[ QH JGVGTQNQIQWU RGEVKPCUG, INWECPCUG, Z[NCPCUG CPF CTCDKPQHWTCPQUKFCUG IGPGU VJGTGHQTG JCXG DGGP GZRTGUUGF KP S. cerevisiae. # RGEVKPQN[VKE YKPG [GCUV YCU FGXGNQRGF D[ EQ-GZRTGUUKPI VJG Erwinia chrysanthemi RGEVCVG N[CUG IGPG ECUUGVVG (PEL5) CPF VJG Erwinia carotovora RQN[ICNCEVWTQPCUG IGPG ECUUGVVG (PEH1) KP S. cerevisiae (öCKPI CPF 2TGVQTKWU 1üüÿ, 1üü3C,D). öKMGYKUG, INWECPQN[VKE YKPG [GCUVU JCXG DGGP FGXGNQRGF D[ GZRTGUUKPI VJG HQNNQYKPI INWECPCUG IGPGU KP S. cerevisiae: VJG Butyrivibrio fibrisolvens GPFQ-¾-1,û-INWECPCUG IGPG (END1); VJG Bacillus subtilis GPFQ-¾-1,3-1,û-INWECPCUG IGPG (BEG1); VJG Ruminococcus flavefaciens EGNNQFGZVTKPCUG IGPG (CEL1); VJG Phanerochaeta chrysosporium EGNNQDKQJ[FTQNCUG IGPG (CBH1); CPF VJG Saccharomycopsis fibuligera EGNNQDKCUG IGPG (BGL1); (2GVGTUGP GV CN. 1üüõ; 8CP 4GPUDWTI GV CN. 1üüû, 1üüó, 1üüù, 1üüø, 1üüõ). ï[NCPQN[VKE [GCUVU YGTG FGXGNQRGF D[ GZRTGUUKPI VJG HQNNQYKPI IGPG ECUUGVVGU KP S. cerevisiae: VJG GPFQ-¾-Z[NCPCUG IGPGU HTQO Aspergillus kawachii (XYN1; %TQWU GV CN. 1üüó); Aspergillus nidulans (xlnA CPF xlnB; 2oTG\-þQP\hNG\ GV CN. 1üüù); Aspergillus niger (XYN4 CPF XYN5; öWVVKI GV CN. 1üüø) CPF Trichoderma reesei (XYN2; öC þTCPIG GV CN. 1üüù, ÿ001); VJG Z[NQUKFCUG IGPGU HTQO A. niger (xlnB) CPF Bacillus pumilus Z[NQUKFCUG (XLO1; öC þTCPIG GV CN. 1üüø); CPF VJG A. niger ½-ö-CTCDKPQHWTCPQUKFCUG IGPG (ABF2; %TQWU GV CN. 1üüù). 6JQWIJ VJG NGXGNU QH UGETGVGF RGEVKPCUGU, INWECPCUGU CPF Z[NCPCUGU CTG UVKNN DGNQY VJG VJTGUJQNF NGXGNU TGSWKTGF HQT VJG GHHGEVKXG TGOQXCN QH RQN[UCEEJCTKFGU
11õ
+UCM 5. 2TGVQTKWU
HTQO ITCRG OWUV CPF YKPG, UQOG FGITGG QH ENCTKHKECVKQP JCU CNTGCF[ DGGP CEJKGXGF. ÷WTVJGTOQTG, RTGNKOKPCT[ HGTOGPVCVKQP VTKCNU JCXG CNUQ KPFKECVGF VJCV UQOG QH VJGUG RQN[UCEEJCTKFG-FGITCFKPI [GCUVU NGCF VQ CP KPETGCUG KP LWKEG [KGNF CPF VJG TGNGCUG QH EQNQWT CPF HNCXQWT EQORQWPFU GPVTCRRGF KP VJG ITCRG UMKPU (8CP 4GPUDWTI CPF 2TGVQTKWU ÿ000). ÷WTVJGT KORTQXGOGPV QH VJGUG [GCUVU EQWNF GXGPVWCNN[ NGCF VQ VJG TGRNCEGOGPV QH QT TGFWEVKQP KP VJG NGXGNU QH EQOOGTEKCN GP\[OG RTGRCTCVKQPU PGGFGF HQT YKPG ENCTKHKECVKQP (2TGVQTKWU ÿ000). 4.4.2.2 Controlled cell flocculation and flotation 9KPGOCMGTU GZRGEV [GCUV EGNNU VQ RGTHQTO VYQ EQPHNKEVKPI TQNGU FWTKPI VJG RTQFWEVKQP QH DQVVNG-HGTOGPVGF URCTMNKPI YKPG (EJCORCIPG) CPF HNQT UJGTT[ (*GPUEJMG 1üüø). &WTKPI VJG RTQFWEVKQP QH VJG DCUG YKPG HQT EJCORCIPG CPF UJGTT[, C JKIJUWURGPFGF [GCUV EQWPV GPUWTGU C TCRKF HGTOGPVCVKQP. +P VJG ECUG QH URCTMNKPI YKPG RTQFWEVKQP, VJG [GCUV EGNNU OWUV HNQEEWNCVG CPF UGVVNG GHHKEKGPVN[ CHVGT VJG EQORNGVKQP QH VJG UGEQPFCT[ HGTOGPVCVKQP KP VJG DQVVNG (prise de mousse). 6JKU HCEKNKVCVGU VJG FGXGNQROGPV QH VJG V[RKECN CWVQN[UKU-OGFKCVGF HGTOGPVCVKQP DQWSWGV CPF OCPPQRTQVGKP-EQCVGF DWDDNGU VJCV CFF VQ VJG HKPGUUG QH VJG URCTMNKPI YKPG CPF CP WPRTQDNGOCVKE TKFFNKPI (rumage) CPF FKUIQTIKPI RTQEGFWTG. +P EQPVTCUV, UJGTT[ RTQFWEVKQP TGSWKTGU [GCUV HNQVCVKQP CPF HNQT (XGNNWO) HQTOCVKQP QP VJG UWTHCEG QH C DCUG YKPG CV VJG GPF QH HGTOGPVCVKQP FWTKPI VJG DKQNQIKECN CIGKPI RTQEGUU (crianza de flor) KP VJG solera U[UVGO CPF VJG FGXGNQROGPV QH VJG EJCTCEVGTKUVKE PWVV[ DQWSWGV (4KDoTGCW-þC[QP GV CN. ÿ000). 6JG EQPVTQNNGF QPUGV QH [GCUV HNQEEWNCVKQP QT HNQT HQTOCVKQP CV VJG CRRTQRTKCVG VKOG ECP UKORNKH[ VJG RTQEGUUKPI QH URCTMNKPI YKPG CPF UJGTT[ (&GSWKP ÿ001). *QYGXGT, VJG IGPGVKE, RJ[UKQNQIKECN CPF DKQEJGOKECN HCEVQTU WPFGTNKPKPI VJG CFCRVKXG TGURQPUGU QH [GCUV EGNNU VQ EGTVCKP RJ[UKECN CPF EJGOKECN UKIPCNU VJCV NGCF VQ HKNCOGPVCVKQP, CIINQOGTCVKQP, HNQEEWNCVKQP CPF HNQVCVKQP CTG PQV HWNN[ WPFGTUVQQF (þCIKCPQ GV CN. ÿ00ÿ). # PWODGT QH FQOKPCPV, UGOK-FQOKPCPV CPF TGEGUUKXG HNQEWNNKP-GPEQFKPI IGPGU (FLO1, FLO2, flo3, FLO4, FLO5, flo6, flo7, FLO9, FLO10 CPF FLO11ñMUC1) CTG KPXQNXGF KP HNQEEWNCVKQP (6GWPKUUGP CPF 5VGGPUOC 1üüó; öCODTGEJVU GV CN. 1üüù; öQ CPF &TCPIKPKU 1üüù). /QTGQXGT, VJG FLO8, MSN1 CPF MSS11 IGPGU GPEQFG TGIWNCVQT[ RTQVGKPU KPXQNXGF KP VJG CEVKXCVKQP QH UQOG QH VJGUG IGPGU, VJGTGD[ RTQOQVKPI VJG HNQEEWNCVKQP QH [GCUV EGNNU (öCODTGEJVU GV CN. 1üüù; 9GDDGT GV CN. 1üüø; %CTUVGPU GV CN 1üüõ; þCIKCPQ GV CN. 1üüüC,D, ÿ00ÿ). FLO1 KU EQPUKFGTGF VQ DG VJG DGUV UVWFKGF CPF RGTJCRU OQUV KORQTVCPV QH CNN VJG HNQEEWNCVKQP IGPGU CPF KU ECRCDNG QH EQPHGTTKPI HNQEEWNCVKQP YJGP VTCPUHQTOGF KPVQ PQP-HNQEEWNGPV S. cerevisiae UVTCKPU (9CVCTK GV CN. 1üü3; $KFCTF GV CN. 1üüû; *COOQPF 1üüù; +UJKFC-÷WLKK GV CN. 1üüõ). +V YCU CNUQ UJQYP VJCV VJG GZRTGUUKQP QH FLO1, NKPMGF VQ VJG NCVGHGTOGPVCVKQP HSP30 RTQOQVGT, ECP DG KPFWEGF D[ C JGCV-UJQEM VTGCVOGPV, EQPHKTOKPI VJCV EQPVTQNNGF HNQEEWNCVKQP FWTKPI HGTOGPVCVKQP KU KPFGGF RQUUKDNG (8GTUVTGRGP GV CN. ÿ001). 6JG JQRG KU VJCV VJKU YKNN NGCF VQ VJG FGXGNQROGPV QH YKPG [GCUV UVTCKPU VJCV HNQEEWNCVG QPN[ WPFGT VJG UVTGUU EQPFKVKQPU (G.I., PWVTKGPV FGRNGVKQP CPF VJG RTGUGPEG QH JKIJ NGXGNU QH CNEQJQN) VJCV WUWCNN[ RTGXCKN CV VJG GPF QH YKPG HGTOGPVCVKQPU (8GTUVTGRGP GV CN. ÿ001).
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
#
11ü
$
%
(KI. ü. (#) # UEJGOCVKE TGRTGUGPVCVKQP QH VJG UVTWEVWTG CPF GP\[OCVKE J[FTQN[UKU QH RGEVKP. ($) # UEJGOCVKE TGRTGUGPVCVKQP QH VJG UVTWEVWTG CPF GP\[OCVKE J[FTQN[UKU QH EGNNWNQUG, EQPUKUVKPI OCKPN[ QH INWECPU. (%) # UEJGOCVKE TGRTGUGPVCVKQP QH VJG UVTWEVWTG CPF GP\[OCVKE J[FTQN[UKU QH JGOKEGNNWNQUG EQPUKUVKPI OCKPN[ QH Z[NCP.
1ÿ0
+UCM 5. 2TGVQTKWU
8GT[ HGY UVTCKPU QH S. cerevisiae CTG ECRCDNG QH ITQYVJ CU C DKQHKNO QP VJG UWTHCEG QH YKPG CPF CTG VJGTGHQTG UWKVCDNG HQT HNQT UJGTT[ RTQFWEVKQP. 6JG IGPQV[RG QH UJGTT[ [GCUVU (HQTOGTN[ MPQYP CU S. beticus CPF S. capensis) VJGTGHQTG KU NKMGN[ VQ DG OQTG EQORNGZ VJCP QTKIKPCNN[ GZRGEVGF. # PWODGT QH IGPGU GPEQFKPI EGNN YCNNCUUQEKCVGF, J[FTQRJQDKE RTQVGKPU JCXG DGGP KORNKECVGF KP XGNNWO HQTOCVKQP. 6JGTG CTG, JQYGXGT, WPEQPHKTOGF, CPGEFQVCN KPFKECVKQPU VJCV VJG IGPGU GPEQFKPI OWEKPNKMG RTQVGKPU CTG KPXQNXGF KP DKQHKNO HQTOCVKQP VJCV TGNCVGU VQ HNQT HQTOCVKQP FWTKPI UJGTT[ CIGKPI. ýPG QH VJGUG IGPGU, FLO11ñMUC1, UGGOU VQ DG C IQQF ECPFKFCVG VQ KPXGUVKICVG HQT VJG RQUUKDKNKV[ QH FGXGNQRKPI UJGTT[ [GCUVU KP YJKEJ XGNNWO HQTOCVKQP ECP DG EQPVTQNNGF. ÷QT GZCORNG, KV KU RQUUKDNG VJCV D[ RNCEKPI VJKU IGPG WPFGT VJG EQPVTQN QH C INWEQUG TGRTGUUKDNG J[DTKF HSP30 RTQOQVGT, VJG HQTOCVKQP QH VJG DKQHKNO EQWNF DG RTQOQVGF CV VJG GPF QH HGTOGPVCVKQP. 4.4.3 +ORTQXGOGPV QH YKPG YJQNGUQOGPGUU 6JG RJTCUG French paradox YCU EQKPGF KP 1üü1 VQ FGUETKDG VJG KPXGTUG TGNCVKQPUJKR DGVYGGP VJG JKIJ KPVCMG QH HCVU KP VJG ÷TGPEJ FKGV CPF VJG NQY KPEKFGPEG QH EQTQPCT[ JGCTV FKUGCUG, CPF JCU CICKP HQEWUGF CVVGPVKQP QP VJG NQPI TGEQIPKUGF DGPGHKV QH OQFGTCVG CNEQJQN EQPUWORVKQP ($KUUQP GV CN. ÿ00ÿ). &WTKPI VJG NCUV FGECFG QT UQ KV JCU DGEQOG YKFGN[ CEEGRVGF VJCV YKPG EQPVCKPU RTQVGEVKXG EQORQWPFU (G.I., RJGPQNKE EQORQWPFU, HNCXQPQKFU, TGUXGTCVTQN, UCNKE[NKE CEKF CPF GVJ[N CNEQJQN) CPF VJCV OQFGTCVG YKPG EQPUWORVKQP QHHGTU UQOG RTQVGEVKQP CICKPUV VJG KPEKFGPEG QH OCP[ FKUGCUGU QH OQFGTP UQEKGV[ (ECTFKQXCUEWNCT FKUGCUG, FKGVCT[ ECPEGTU, KUEJCGOKE UVTQMG, RGTKRJGTCN XCUEWNCT FKUGCUG, FKCDGVGU, J[RGTVGPUKQP, RGRVKE WNEGTU, MKFPG[ UVQPGU CPF OCEWNCT FGIGPGTCVKQP) CPF UVKOWNCVGU TGUKUVCPEG VQ KPHGEVKQP CPF TGVGPVKQP QH DQPG FGPUKV[ (#TOUVTQPI GV CN. ÿ001; $KUUQP GV CN. ÿ00ÿ). 2TWFGPV YKPG FTKPMGTU CTG KPETGCUKPIN[ HCUVKFKQWU CDQWV VJG RTGUGPEG QH WPFGUKTCDNG EQORQWPFU KP YKPG. 6JGUG WPYCPVGF EQORQWPFU KPENWFG UWURGEVGF ECTEKPQIGPU UWEJ CU GVJ[N ECTDCOCVG, PGWTQVQZKPU UWEJ CU DKQIGPKE COKPGU, CPF CUVJOCVKE EJGOKECN RTGUGTXCVKXGU UWEJ CU UWNRJKVGU (&W 6QKV CPF 2TGVQTKWU ÿ000). 5QOG YKPG FTKPMGTU CTG CNUQ EQPEGTPGF CDQWV JKIJ NGXGNU QH CNEQJQN KP YKPG. 9JGP YKPG [GCUV UVTCKPU CTG FGXGNQRGF, KV KU QH VJG WVOQUV KORQTVCPEG VQ HQEWU QP VJGUG JGCNVJ CURGEVU CPF VQ FGXGNQR [GCUVU VJCV OC[ GPJCPEG VJG DGPGHKVU (G.I., RTQFWEKPI TGUXGTCVTQN) CPF TGFWEG VJG TKUMU (G.I., GNKOKPCVKPI GVJ[N ECTDCOCVG CPF DKQIGPKE COKPGU, CPF TGFWEKPI JKIJ NGXGNU QH CNEQJQN) CUUQEKCVGF YKVJ OQFGTCVG YKPG EQPUWORVKQP (&W 6QKV CPF 2TGVQTKWU ÿ000; 2TGVQTKWU ÿ000, ÿ003). 4.4.3.1 Increased production of resveratrol þTCRG RJ[VQCNGZKPU, KPENWFKPI UVKNDGPGU UWEJ CU TGUXGTCVTQN, PQV QPN[ TGFWEG VJG TKUM QH EQTQPCT[ JGCTV FKUGCUG, DWV CNUQ CEV CU CPVKQZKFCPVU CPF CPVKOWVCIGPU (#TOUVTQPI GV CN. ÿ001). 6JG[ FKURNC[ CPVK-KPHNCOOCVQT[ CPF ECPEGT EJGOQRTGXGPVKXG RTQRGTVKGU, CU YGNN CU VJG CDKNKV[ VQ KPFWEG URGEKHKE GP\[OGU VJCV OGVCDQNKUG ECTEKPQIGPKE UWDUVCPEGU. 4GUXGTCVTQN KU U[PVJGUKUGF OCKPN[ KP VJG UMKP EGNNU QH ITCRG DGTTKGU; QPN[ VTCEGU CTG HQWPF KP VJG HNGUJ. #U C TGUWNV, TGF YKPG JCU C OWEJ JKIJGT
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
1ÿ1
TGUXGTCVTQN EQPEGPVTCVKQP VJCP YJKVG YKPG FWG VQ UMKP EQPVCEV FWTKPI VJG HKTUV RJCUG QH HGTOGPVCVKQP. +P CP CVVGORV VQ KPETGCUG VJG NGXGNU QH TGUXGTCVTQN KP DQVJ TGF CPF YJKVG YKPG, VJG RJGP[NRTQRCPQKF RCVJYC[ KP S. cerevisiae JCU DGGP OQFKHKGF VQ RTQFWEG ÐEQWOCTQ[N-EQGP\[OG #, QPG QH VJG UWDUVCPEGU HQT TGUXGTCVTQN U[PVJGUKU ($GEMGT ÿ00ÿ). 6JG QVJGT UWDUVTCVG, OCNQP[N-EQGP\[OG #, KU CNTGCF[ HQWPF KP [GCUV CPF KU KPXQNXGF KP de novo HCVV[ CEKF DKQU[PVJGUKU. +V KU J[RQVJGUKUGF VJCV VJG RTQFWEVKQP QH Ð-EQWOCTQ[N-EQGP\[OG # ECP DG CEJKGXGF D[ KPVTQFWEKPI VJG RJGP[NCNCPKPG COOQPKC-N[CUG IGPG (PAL), VJG EKPPCOCVG û-J[FTQZ[NCUG IGPG (D4H), VJG EQGP\[OG # NKICUG IGPG (4CL9 CPF 4CL216) CPF VJG ITCRG UVKNDGPG U[PVJCUG IGPG (Vst1) KPVQ S. cerevisiae. 9JGP KPVTQFWEGF, VJG ITCRG UVKNDGPG U[PVJCUG IGPG OC[ VJGP ECVCN[UG VJG CFFKVKQP QH VJTGG CEGVCVG WPKVU HTQO OCNQP[N-EQGP\[OG # VQ pEQWOCT[N-EQGP\[OG #, TGUWNVKPI KP VJG HQTOCVKQP QH TGUXGTCVTQN. +P C RTGNKOKPCT[ UVWF[, VJG 4CL9, 4CL216 CPF Vst1 IGPGU YGTG ENQPGF WPFGT VJG EQPVTQN QH UVTQPI [GCUV RTQOQVGTU QH VJG CNEQJQN FGJ[FTQIGPCUG ++ (ADH2) CPF GPQNCUG (ENO2) IGPGU CPF VTCPUHQTOGF KPVQ S. cerevisiae ($GEMGT ÿ00ÿ). +PKVKCNN[, PQ TGUXGTCVTQN EQWNF DG FGVGEVGF KP VJG [GCUV GZRTGUUKPI VJG EQGP\[OG # NKICUG CPF TGUXGTCVTQN U[PVJCUG IGPGU. ýPN[ CHVGT VJG UCORNGU YGTG VTGCVGF YKVJ ¾-INWEQUKFCUG VQ TGNGCUG VJG INWEQUG OQKGVKGU HTQO VJG UQ-ECNNGF RKEGKF OQNGEWNG (VJG INWEQUKFG HQTO QH TGUXGTCVTQN) YGTG FGVGEVCDNG NGXGNU RGTEGKXGF KP OCUU URGEVTQOGVTKE CUUC[U. 6JKU JCU GPEQWTCIGF GHHQTVU VQ EQ-GZRTGUU VJG EQGP\[OG # NKICUG CPF TGUXGTCVTQN U[PVJCUG IGPGU, VQIGVJGT YKVJ C INWEQUG-TGUKUVCPV ¾-INWEQUKFCUG IGPG UWEJ CU VJG BGL1 IGPG HTQO S. fibuligera. /QTG KPXGUVKICVKQP KP ITGCVGT FGRVJ OWUV DG EQPFWEVGF DGHQTG KV ECP DG TGEQOOGPFGF CU C YC[ VQ FGXGNQR TGUXGTCVTQN-RTQFWEKPI YKPG [GCUV UVTCKPU. 4.4.3.2 Reduced formation of ethyl carbamate ©VJ[N ECTDCOCVG (C ECTEKPQIGPKE EQORQWPF CNUQ MPQYP CU WTGVJCPG) KP YKPGU CPF TGNCVGF RTQFWEVU HQTOU HTQO VJG TGCEVKQP DGVYGGP WTGC CPF GVJCPQN (ýWIJ GV CN. 1üõõ). +P S. cerevisiae, WTGC KU RTQFWEGF D[ VJG CAR1-GPEQFGF CTIKPCUG FWTKPI VJG DTGCMFQYP QH CTIKPKPG, QPG QH VJG OCKP COKPQ CEKFU KP ITCRG LWKEG. +P VJKU RTQEGUU, CTIKPKPG KU EQPXGTVGF VQ QTPKVJKPG, COOQPKC CPF ECTDQP FKQZKFG, CPF WTGC KU HQTOGF CU CP KPVGTOGFKCVG RTQFWEV. 7TGC KU UGETGVGF KPVQ VJG YKPG D[ EGTVCKP [GCUV UVTCKPU VJCV, FGRGPFKPI QP HGTOGPVCVKQP EQPFKVKQPU, OC[ DG WPCDNG VQ HWTVJGT OGVCDQNKUG VJG GZVGTPCN WTGC. 6JQWIJ CNN S. cerevisiae UVTCKPU UGETGVG WTGC, VJG GZVGPV VQ YJKEJ VJG[ TG-CDUQTD VJG WTGC FKHHGTU (#P CPF ýWIJ 1üü3). öQY WTGC-HQTOKPI [GCUV UVTCKPU CTG UGNGEVGF CPF EQOOGTEKCN RTGRCTCVKQPU QH CEKFKE WTGCUG CTG CFFGF YKVJ UQOG UWEEGUU VQ TGFWEG VJG CEEWOWNCVKQP QH WTGC KP YKPG. 6Q EWTD VJG HQTOCVKQP QH GVJ[N ECTDCOCVG, VJG UWEEGUUKXG FKUTWRVKQP QH VJG CAR1 CTIKPCUG IGPG KP CP KPFWUVTKCN UCMo [GCUV UWEEGUUHWNN[ GNKOKPCVGF WTGC CEEWOWNCVKQP KP TKEG YKPG (îKVCOQVQ GV CN. 1üü1). 6JKU CTIKPCUG FGNGVKQP OWVCVKQP NGF VQ C [GCUV UVTCKP VJCV EQWNF PQV OGVCDQNKUG CTIKPKPG. *QYGXGT, ITQYVJ YCU CNUQ KORGFGF, YJKEJ NKOKVU VJG EQOOGTEKCN WUG QH VJG UVTCKP. +P CPQVJGT CVVGORV VQ GNKOKPCVG WTGC CU VJG RTGEWTUQT QH GVJ[N ECTDCOCVG HTQO YKPG, C WTGCUG IGPG YCU EQPUVTWEVGF D[ HWUKPI VJG ½, ¾ CPF Ä UWDWPKVU QH VJG Lac-
1ÿÿ
+UCM 5. 2TGVQTKWU
tobacillus fermentum WTGCUG QRGTQP, KPUGTVKPI VJG LCEM DGCP WTGCUG NKPMGT UGSWGPEGU DGVYGGP DQVJ VJG ½ CPF ¾ CPF VJG ¾ CPF Ä UWDWPKVU, CPF NKPMKPI VJKU EQPUVTWEV VQ VJG [GCUV PGK1 RTQOQVGT (8KUUGT 1üüü). *QYGXGT, VJG UGETGVKQP QH WTGCUG RGRVKFGU YCU GZVTGOGN[ NQY CPF WPCDNG VQ EQPXGTV WTGC KPVQ COOQPKC CPF ECTDQP FKQZKFG. 6JG CDUGPEG QH DKQNQIKECN CEVKXKV[ KP VJG TGEQODKPCPV WTGCUG YCU RTQDCDN[ VJG TGUWNV QH C NCEM QH VJG GUUGPVKCN CWZKNKCT[ RTQVGKPU, YJKEJ CTG RTGUGPV QPN[ KP WTGCUG-RTQFWEKPI URGEKGU, UWEJ CU VJG HKUUKQP [GCUV Schizosaccharomyces pombe. 9KVJQWV VJGUG RTQVGKPU, S. cerevisiae KU WPCDNG VQ CUUGODNG VJG XCTKQWU UWDWPKVU KPVQ CP CEVKXG WTGCUG. +V UGGOU, VJGTGHQTG, VJCV HQT S. cerevisiae VQ GZRTGUU CP CEVKXG WTGCUG, CEEGUUQT[ IGPGU QH L. fermentum YKNN CNUQ JCXG VQ DG ENQPGF CPF GZRTGUUGF KP CFFKVKQP VQ VJG UVTWEVWTCN WTGCUG IGPGU. 4. 4.3.3 Reduced formation of biogenic amines $KQIGPKE COKPGU QTKIKPCVG OCKPN[ HTQO VJG FGECTDQZ[NCVKQP QH RTGEWTUQT COKPQ CEKFU D[ UWDUVTCVG-URGEKHKE GP\[OGU QH VJG DCEVGTKC (KPENWFKPI VJG OCNQNCEVKE DCEVGTKC) KP YKPG. *KUVKFKPG, V[TQUKPG, RJGP[NCNCPKPG CPF N[UKPG CTG VJG RTGEWTUQTU QH JKUVCOKPG, V[TCOKPG, RJGPGVJ[NCOKPG CPF ECFCXGTKPG TGURGEVKXGN[, YJGTGCU RQN[COKPGU, UWEJ CU RWVTGUEKPG, URGTOKPG CPF URGTOKFKPG, CTG U[PVJGUKUGF HTQO QTPKVJKPG CPF CTIKPKPG (6QTTGC þQwK CPF #PEsP #\RKNKEWGVC ÿ001). 6JG KPIGUVKQP QH RTQFWEVU EQPVCKPKPI TGNCVKXGN[ JKIJ NGXGNU QH DKQIGPKE COKPGU ECP RTQXQMG CFXGTUG GHHGEVU, UWEJ CU J[RQVGPUKQP, HNWUJKPI, JGCFCEJG CPF OKITCKPG. 6JG COKPG IGPGTCNN[ RTGUGPV KP VJG JKIJGUV EQPEGPVTCVKQP KP YKPG KU RWVTGUEKPG, CPF KV KU CNUQ MPQYP CU VJG OQUV GHHGEVKXG RQVGPVKCVQT QH JKUVCOKPG VQZKEKV[ KP JWOCPU (þWGTTKPK GV CN. ÿ00ÿ). /QTGQXGT, CU C DKQIGPKE COKPG RWVTGUEKPG, KV KU C RQVGPVKCN RTGEWTUQT QH ECTEKPQIGPKE PKVTQUCOKPGU. 6JG RTGUGPEG QH DKQIGPKE COKPGU KP YKPG KU VJGTGHQTG KPETGCUKPIN[ DGEQOKPI C JGCNVJ EQPEGTP. # TGEGPV KPXGUVKICVKQP QH VJG KPHNWGPEG QH FKHHGTGPV UVTCKPU QH S. cerevisiae QP VJG EQPEGPVTCVKQP QH DKQIGPKE COKPGU KP YKPG TGXGCNGF VJCV HQT VJG XCTKQWU UVTCKPU VGUVGF VJGTG YGTG QPN[ UNKIJV FKHHGTGPEGU KP VJG EQPVGPV QH VJG COKPGU, CPF JKIJ EQPEGPVTCVKQPU YGTG PGXGT TGCEJGF (6QTTGC þQwK CPF #PEsP #\RKNKEWGVC ÿ001). /QTGQXGT, PQ TGNCVKQPUJKR YCU HQWPF DGVYGGP VJG EQPEGPVTCVKQP QH DKQIGPKE COKPGU KP YKPG CPF VJG WVKNKUCVKQP QH VJGKT RTGEWTUQT COKPQ CEKFU D[ VJG FKHHGTGPV UVTCKPU QH S. cerevisiae FWTKPI VJG CNEQJQNKE HGTOGPVCVKQP. 6JKU EQPHKTOGF VJG PQVKQP VJCV DKQIGPKE COKPGU KP YKPG CTG RTQFWEGF OCKPN[ D[ URQKNCIG DCEVGTKC (G.I., Lactobacillus, Leuconostoc CPF Pediococcus), CU YGNN CU D[ UQOG UVTCKPU QH VJG OCNQNCEVKE HGTOGPVCVKQP DCEVGTKWO (Oenococcus oeni) (þWGTTKPK GV CN. ÿ00ÿ). # NQIKECN CRRTQCEJ VQ TGFWEKPI VJG NGXGNU QH DKQIGPKE COKPGU KP YKPG UGGOU VQ DG VJG FGXGNQROGPV QH DCEVGTKEKFCN YKPG [GCUV UVCTVGT EWNVWTG UVTCKPU VJCV YQWNF EWTD VJG ITQYVJ QH NCEVKE CEKF DCEVGTKC. 6JG EQPUVTWEVKQP QH YKPG [GCUVU VJCV UGETGVG C YKFG URGEVTWO QH CPVKDCEVGTKCN EQORQWPFU YQWNF WPFQWDVGFN[ EQPVTKDWVG VQ VJG KPJKDKVKQP QH DKQCOKPG-RTQFWEKPI NCEVKE CEKF DCEVGTKC KP YKPG (5EJQGOCP GV CN. 1üüü; &W 6QKV CPF 2TGVQTKWU ÿ000; &W 6QKV GV CN. ÿ003). 6JG FGXGNQROGPV QH C OCNQNCEVKE YKPG [GCUV UVTCKP JCU CNUQ DGGP RTQRQUGF CU CP CNVGTPCVKXG VQ VJG WUG QH NCEVKE CEKF DCEVGTKC UWEJ CU O. oeni VQ EQPFWEV VJG OCNQNCEVKE HGTOGPVCVKQP (UGG UGEVKQP û.û.ó). 6JKU YQWNF CXQKF VJG TKUM QH DCEVGTKCNN[ RTQFWEGF DKQIGPKE COKPGU
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
1ÿ3
KP YKPG (9KNNKCOU GV CN. 1üõû; &GPC[TQNNGU GV CN. 1üüó; #PUCPC[ GV CN. 1üü3, 1üüù; öCDCTTG GV CN. 1üüù; $QP[ GV CN. 1üüø; 8QNUEJGPM GV CN. 1üüø). 4.4.3.4 Increased production of biopreservative compounds and enzymes 7PEQPVTQNNGF OKETQDKCN ITQYVJ DGHQTG, FWTKPI QT CHVGT YKPG HGTOGPVCVKQP ECP CNVGT VJG EJGOKECN EQORQUKVKQP QH VJG GPF RTQFWEV, CPF FGVTCEV HTQO CRRGCTCPEG, CTQOC CPF HNCXQWT. *GCNVJ[ ITCRGU, EGNNCT J[IKGPG CPF UQWPF QGPQNQIKECN RTCEVKEGU CTG VJG YKPGOCMGTU•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ýÿ FWTKPI YKPG HGTOGPVCVKQPU), CPVKOKETQDKCN GP\[OGU (G.I., N[UQ\[OG, EJKVKPCUGU, GPFQINWECPCUGU, GVE.) CPF RGRVKFGU (\[OQEKPU CPF DCEVGTKQEKPU) KU EWTTGPVN[ DGKPI EQPUKFGTGF CU CP CNVGTPCVKXG VQ EJGOKECN RTGUGTXCVKQP. $WV VJG WUG QH EQOOGTEKCN CPVKOKETQDKCN GP\[OG RTGRCTCVKQPU CPF DCEVGTKQEKPU KU GZRGPUKXG; VJKU RTQDNGO OKIJV DG EKTEWOXGPVGF D[ GZRTGUUKPI GHHGEVKXG CPVKOKETQDKCN GP\[OGU CPF RGRVKFGU KP YKPG [GCUV UVCTVGT EWNVWTG UVTCKPU. #PVKOKETQDKCN GP\[OGU, UWEJ CU EJKVKPCUGU, ¾-INWECPCUGU CPF N[UQ\[OG, CEV D[ FGITCFKPI MG[ UVTWEVWTCN EQORQPGPVU QH VJG EGNN YCNNU QH OQWNFU CPF DCEVGTKC. %JKVKPCUGU CPF ¾-INWECPCUGU U[PGTIKUVKECNN[ CVVCEM EJKVKP CPF ¾-1,3-INWECP, VJG OCKP EQORQPGPVU QH HWPICN EGNN YCNNU. 6JG N-CEGV[NJGZQUCOKPKFCUG N[UQ\[OG, QP VJG QVJGT JCPF, N[UGU VJG EGNN YCNNU QH þTCO-RQUKVKXG URGEKGU QH DCEVGTKC VJCV NCEM CP QWVGT OGODTCPG D[ J[FTQN[UKPI VJG ¾-1,û-INWEQUKFKE NKPMCIGU QH RGRVKFQIN[ECP KP VJG EGNN YCNN (÷WINUCPI GV CN. 1üüó; %CTUVGPU GV CN. ÿ003). #NVJQWIJ EQOOGTEKCN RTGRCTCVKQPU QH CPVKOKETQDKCN GP\[OGU UWEJ CU N[UQ\[OG JCXG DGGP CRRTQXGF VQ EQPVTQN VJG ITQYVJ QH URQKNCIG QTICPKUOU, VJG[ CTG PQV YKFGN[ WUGF KP YKPGOCMKPI DGECWUG QH VJGKT NQY EQUV GHHKEKGPE[. 6JKU JCU OQVKXCVGF GHHQTVU VQ FGXGNQR S. cerevisiae UVTCKPU RTQFWEKPI VJGUG CPVKOKETQDKCN GP\[OGU, UWEJ CU EJKVKPCUGU, ¾-INWECPCUGU CPF N[UQ\[OG (òCMCOWTC GV CN. 1üü3; %CTUVGPU GV CN. ÿ003). 5QOG UWEEGUU JCU CNTGCF[ DGGP CEJKGXGF YKVJ VJG FGXGNQROGPV QH CPVKHWPICN [GCUVU KP YJKEJ VJG CTS1 EJKVKPCUG CPF EXG1 GPFQINWECPCUG IGPGU CTG GZRTGUUGF (%CTUVGPU GV CN. ÿ003). ÷WTVJGTOQTG, VJG N[UQ\[OG-GPEQFKPI IGPG (HEL1) HTQO EJKEMGP GII YJKVG YCU UWEEGUUHWNN[ GZRTGUUGF KP S. cerevisiae CPF VJG DCEVGTKEKFCN CEVKQP QH VJG TGEQODKPCPV N[UQ\[OG CICKPUV þTCO-PGICVKXG DCEVGTKC YCU GPJCPEGF YJGP C RGPVCRGRVKFG YCU KPUGTVGF KPVQ KVU %-VGTOKPWU (+DTCJKO GV CN. 1üüÿ). 4GUGCTEJ KU DGKPI FQPG VQ GZRTGUU C OQFKHKGF N[UQ\[OG IGPG KP YKPG [GCUV VJCV YQWNF CXQKF J[RGTIN[EQU[NCVKQP CPF DTQCFGP KVU CEVKXKV[ VQ GHHGEVKXGN[ GNKOKPCVG URQKNCIG D[ NCEVKE CPF CEGVKE CEKF DCEVGTKC.
1ÿû
+UCM 5. 2TGVQTKWU
7PNKMG VJG FKTGEV GHHGEV QH VJG CPVKHWPICN [GCUV CTS1-GPEQFGF EJKVKPCUG CPF EXG1-GPEQFGF GPFQINWECPCUG (%CTUVGPU GV CN. ÿ003) CPF VJG CPVKDCEVGTKCN N[UQ\[OG-GPEQFKPI IGPG HTQO EJKEMGP GII YJKVG, UQOG GP\[OGU JCXG CP KPFKTGEV GHHGEV QP URQKNCIG OKETQDGU. ÷QT GZCORNG, VJG GZRTGUUKQP QH VJG A. niger INWEQUG QZKFCUG IGPG ECUUGVVG (GOX1) KP S. cerevisiae TGUWNVGF KP VJG RTQFWEVKQP QH J[FTQIGP RGTQZKFG. 6JKU, KP VWTP, NGF VQ J[RGTDCTKE QZ[IGP VQZKEKV[, C TGUWNV QH VJG RGTQZKFCVKQP QH DCEVGTKCN OGODTCPG NKRKFU, CPF C UVTQPI QZKFKUKPI GHHGEV QP VJG DCEVGTKCN EGNNU, VJG ECWUG QH VJG FGUVTWEVKQP QH DCUKE OQNGEWNCT UVTWEVWTGU UWEJ CU PWENGKE CEKFU CPF EGNN RTQVGKPU (/CNJGTDG GV CN. ÿ00ÿ). +V TGOCKPU VQ DG GUVCDNKUJGF YJGVJGT VJG NGXGN QH GOX1-IGPGTCVGF J[FTQIGP RGTQZKFG KP VJG HKPKUJGF YKPG YQWNF DG WPFGT VJG NKOKV UGV D[ CWVJQTKVKGU. %WTTGPVN[ EQPUKFGTCVKQP KU DGKPI IKXGP VQ VJG NCVG GZRTGUUKQP QH C ECVCNCUG IGPG KP YKPG [GCUV, YJKEJ ECP GNKOKPCVG GZEGUUKXG GOX1IGPGTCVGF J[FTQIGP RGTQZKFG CHVGT KV JCU RNC[GF C RTGUGTXCVKXG TQNG FWTKPI VJG GCTNKGT RJCUGU QH YKPGOCMKPI. <[OQEKPU RTQFWEGF D[ S. cerevisiae CTG NGVJCN QPN[ VQ UGPUKVKXG UVTCKPU QH S. cerevisiae (UGG 5GEVKQP û.1.3), YJGTGCU VJQUG RTQFWEGF D[ PQP-Saccharomyces URGEKGU OC[ DG VQZKE VQ PQP-Saccharomyces URGEKGU CU YGNN (5JKOK\W 1üü3). 6JGTG JCXG DGGP UGXGTCN CVVGORVU VQ GZRCPF VJG \[OQEKFCN CEVKXKV[ QH S. cerevisiae UQ VJCV KV EQWNF CNUQ GNKOKPCVG EQPVCOKPCVKPI URQKNCIG YKPG [GCUVU KP ITCRG OWUV CPF YKPG. 8CTKQWU MKNNGT V[RGU (G.I., î1 CPF îÿõ) QH S. cerevisiae JCXG DGGP J[DTKFKUGF D[ OCVKPI, E[VQFWEVKQP CPF URJGTQRNCUV HWUKQP CPF, KP QPG ECUG, C &ò# EQR[ QH VJG î1 FU4ò# YCU KPVGITCVGF KPVQ VJG IGPQOG QH C îÿ UVTCKP QH S. cerevisiae ($QQPG GV CN. 1üü0). *QYGXGT, FWG VQ VJG PCTTQY URGEVTWO QH MKNNKPI CPF VJG HCEV VJCV VJG î1 MKNNGT VQZKP KU KPGHHGEVKXG CV VJG NQY R* QH YKPG, VJKU î1ñîÿ FQWDNG MKNNGT S. cerevisiae KU PQV WUGHWN HQT EQPVTQNNKPI VJG YKFG XCTKGV[ QH [GCUV EQPVCOKPCPVU FWTKPI YKPG HGTOGPVCVKQPU. 6JGTGHQTG, VJG HQEWU KU PQY QP VJG KFGPVKHKECVKQP QH IGPGU GPEQFKPI OQTG GHHGEVKXG \[OQEKPU CPF DCEVGTKQEKPU CPF VJGKT RQUUKDNG KPVTQFWEVKQP KPVQ YKPG [GCUVU. $CEVGTKQEKPU UWEJ CU PKUKP, RGFKQEKP CPF NGWEQEKP JCXG DGGP UJQYP VQ UWRRTGUU VJG RTQNKHGTCVKQP QH UGXGTCN þTCO-RQUKVKXG URQKNCIG DCEVGTKC KP YKPG (&W 6QKV GV CN. ÿ003). #NVJQWIJ VJG[ RTQXKFG VJG YKPGOCMKPI EQOOWPKV[ YKVJ C UCHG CNVGTPCVKXG VQ EJGOKECN RTGUGTXCVKQP, VJGKT NQY EQUV GHHKEKGPE[ KU C NKOKVKPI HCEVQT. 6Q FGCN YKVJ VJG EQUV HCEVQT, TGUGCTEJGTU JCXG NQQMGF CV VJG HGCUKDKNKV[ QH EQPVTQNNKPI URQKNCIG DCEVGTKC FWTKPI YKPG HGTOGPVCVKQPU D[ GPIKPGGTKPI DCEVGTKEKFCN UVTCKPU QH S. cerevisiae. 6Q VJKU GPF, VJG IGPGU GPEQFKPI VJG Pediococcus acidilactici RGFKQEKP (PED1) CPF VJG Leuconostoc carnosum NGWEQEKP (LCA1) YGTG ENQPGF WPFGT VJG EQPVTQN QH VJG ADH1 IGPG RTQOQVGT CPF MF½1 UGETGVKQP UKIPCN UGSWGPEGU CPF GZRTGUUGF KP NCDQTCVQT[ UVTCKPU QH S. cerevisiae (5EJQGOCP GV CN. 1üüü). $KQNQIKECNN[ CEVKXG CPVKOKETQDKCN RGRVKFGU RTQFWEGF D[ VJG S. cerevisiae VTCPUHQTOCPVU YGTG KPFKECVGF VQ DG RTGUGPV D[ CICT FKHHWUKQP CUUC[U CICKPUV UGPUKVKXG KPFKECVQT DCEVGTKC, UWEJ CU VJG YGNN-MPQYP HQQF RCVJQIGP, Listeria monocytogenes, CPF VJG HGTQEKQWU YKPG URQKNCIG DCEVGTKWO, Lactobacillus kunkeei. 6JG JGVGTQNQIQWU RGRVKFGU YGTG RTGUGPV CV TGNCVKXGN[ NQY NGXGNU KP VJG EWNVWTG UWRGTPCVCPV, DWV RGFKQEKP CPF NGWEQEKP CEVKXKVKGU YGTG FGVGEVGF TGCFKN[ YJGP KPVCEV [GCUV EQNQPKGU YGTG WUGF KP UGPUKVKXG UVTCKP QXGTNC[U. 6JGUG RTQOKUKPI RTGNKOKPCT[ TGUWNVU GPEQWTCIGF HWTVJGT KPXGUVKICVKQPU, YJKEJ HQEWU QP VJG KPETGCUG KP VJG GZVTCEGNNWNCT NGXGNU QH RGFKQEKP CPF NGWEQEKP KP S. cerevisiae CPF VJG EQ-GZRTGUUKQP QH PED1 CPF LCA1 VQIGVJGT
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
1ÿó
NGWEQEKP KP S. cerevisiae CPF VJG EQ-GZRTGUUKQP QH PED1 CPF LCA1 VQIGVJGT YKVJ IGPGU GPEQFKPI QVJGT DCEVGTKQEKPU CPF DCEVGTKQN[VKE GP\[OGU. 6JKU CRRTQCEJ YKNN DTQCFGP VJG URGEVTWO QH WPYCPVGF DCEVGTKC VJCV YQWNF DG KPJKDKVGF D[ CP GPIKPGGTGF YKPG [GCUV UVTCKP, CPF RCXG VJG YC[ HQT VJG RTQFWEVKQP QH YKPG YKVJ TGFWEGF NGXGNU QH RQVGPVKCNN[ JCTOHWN EJGOKECN RTGUGTXCVKXGU. 4.4.3.5 Decreased levels of alcohol *GCNVJ EQPUEKQWUPGUU CPF KPETGCUKPIN[ UVTKEV VTCHHKE NCYU RGTVCKPKPI VQ ÉFTKPMKPI CPF FTKXKPIÌ UGGO VQ DG VJG OCKP TGCUQPU HQT C YQTNFYKFG FGENKPG KP VJG EQPUWORVKQP QH CNEQJQN. 6JKU JCU KPETGCUGF VJG FGOCPF HQT NQY CNEQJQN, TGFWEGF CNEQJQN CPF FG-CNEQJQNKUGF YKPGU, RWVVKPI C ITGCV FGCN QH RTGUUWTG QP YKPG RTQFWEGTU, RCTVKEWNCTN[ VJQUG KP YCTO YKPG-RTQFWEKPI TGIKQPU YJGTG ITCRG UWICT NGXGNU CTG OWEJ JKIJGT (5EWFCOQTG-5OKVJ CPF /QTCP 1üüø; 2KEMGTKPI GV CN. 1üüõ). +P TGURQPUG VQ VJKU OCTMGV EJCPIG, UGXGTCN GZRGPUKXG RJ[UKECN CPF EJGOKECN RTQEGUUGU (UQOGVKOGU WUGF KP EQODKPCVKQP) JCXG DGGP FGXGNQRGF HQT VJG RCTVKCN TGOQXCN QH CNEQJQN HTQO YKPG. 6JGUG RTQEGUUGU (UQOG QH YJKEJ OKIJV CNUQ DG FGNGVGTKQWU VQ VJG SWCNKV[ QH VJG GPF RTQFWEV) KPENWFG VJGTOCN GXCRQTCVKQP, FKUVKNNCVKQP, OGODTCPG HKNVTCVKQP, CFUQTRVKQP, EGPVTKHWICVKQP, HTGG\G EQPEGPVTCVKQP CPF RCTVKCN HGTOGPVCVKQP (2KEMGTKPI CPF *GCVJGTDGNN 1üüù; 2KEMGTKPI 1üüõ; 2KEMGTKPI GV CN. 1üüõ, 1üüüC,D, E). 6JG NGXGNU QH CNEQJQN KP HGTOGPVGF DGXGTCIGU ECP DG TGFWEGF D[ TGFKTGEVKPI VJG ECTDQP HNWZ CYC[ HTQO GVJCPQN HQTOCVKQP CPF VQYCTFU VJG RTQFWEVKQP QH IN[EGTQN CPF INWEQPKE CEKF. # UKIPKHKECPV KPETGCUG KP VJG NGXGN QH GZVTCEGNNWNCTN[ CEEWOWNCVGF IN[EGTQN CPF EQPEQOKVCPV FGETGCUGU KP GVJCPQN EQPEGPVTCVKQPU JCXG DGGP CEJKGXGF D[ VJG QXGTGZRTGUUKQP QH VJG GPFQIGPQWU GPD1- CPF GPD2-GPEQFGF IN[EGTQN-3-RJQURJCVG FGJ[FTQIGPCUG KUQ\[OGU QH S. cerevisiae, VQIGVJGT YKVJ VJG EQPUVKVWVKXG GZRTGUUKQP QH KVU FPS1-GPEQFGF IN[EGTQN VTCPURQTV HCEKNKVCVQT. 5KOKNCT FGETGCUGU KP GVJCPQN NGXGNU JCXG DGGP CEJKGXGF D[ VJG GZRTGUUKQP QH VJG Aspergillus niger GOX1 INWEQUG QZKFCUG IGPG KP S. cerevisiae (/KEJPKEM GV CN. 1üüø; 4GOK\G GV CN. 1üüü; &G $CTTQU öQRGU GV CN. ÿ000; ©INKPVQP GV CN. ÿ00ÿ; /CNJGTDG GV CN. ÿ00ÿ). +V TGOCKPU VQ DG RTQXGP YJGVJGT VJG JGVGTQNQIQWU GZRTGUUKQP QH VJG A. niger GOX1 IGPG, VQIGVJGT YKVJ VJG QXGTGZRTGUUKQP QH VJG S. cerevisiae GPD1, GPD2 CPF FPS1 IGPGU KP YKPG [GCUV UVTCKPU, EQWNF RTQXKFG C XKCDNG OGCPU QH NQYGTKPI VJG CNEQJQN EQPVGPV VQ FGUKTGF NGXGNU KP EQOOGTEKCN YKPGU. 4.4.4 +ORTQXGOGPV QH YKPG HNCXQWT CPF QVJGT UGPUQT[ SWCNKVKGU +P OQFGTP YKPGOCMKPI, EQPUWOGT QNHCEVQT[ RTQHKNKPI KU DGEQOKPI KPETGCUKPIN[ KORQTVCPV CU C IWKFG VQ RTQFWEVKQP FGEKUKQPU ($KUUQP GV CN. ÿ00ÿ). 6JG EQODKPGF WUG QH EJGOKECN CPF UGPUQT[ CPCN[UGU QH YKPG PQY OCMGU KV RQUUKDNG VQ WPFGTUVCPF VJG UWDVNG PWCPEGU CUUQEKCVGF YKVJ XCTKGVCN YKPG HNCXQWT CPF HGTOGPVCVKQP DQWSWGV. 6JG QTICPQNGRVKE SWCNKV[ (CRRGCTCPEG, CTQOC CPF HNCXQWT) QH YKPG FGRGPFU QP VJG RTGUGPEG QH CDUQNWVG COQWPVU CPF URGEKHKE TCVKQU QH OCP[ FGUKTCDNG CPF KPVGTCEVKXG EQORQWPFU, CPF VJG CDUGPEG QH PGICVKXG QPGU. 5WDVNG EQODKPCVKQPU QH VTCEG ITCRG-
1ÿù
+UCM 5. 2TGVQTKWU
FGTKXGF EQORQPGPVU WUWCNN[ GNKEKV VJG EJCTCEVGTKUVKE HNCXQWT CPF CTQOC PQVGU QH YKPG; VJGUG EQORQPGPVU KPENWFG CEEWOWNCVGF UGEQPFCT[ OGVCDQNKVGU UWEJ CU VGTRGPGU KP VJG CTQOCVKE XCTKGVKGU CPF CNMQZ[R[TC\KPGU KP VJG XGIGVCVKXG QT JGTDCEGQWU EWNVKXCTU. 6JG RTQFWEVU QH [GCUV HGTOGPVCVKQP (G.I., GUVGTU, CNEQJQNU, CNFGJ[FGU, FKCEGV[N, HCVV[ CEKFU, RJGPQNKE CEKFU, UWNRJKVGU CPF UWNRJKFGU, GVE.) EQPVTKDWVG VQ VJG IGPGTKE DCEMITQWPF HNCXQWT CPF CTQOC, CU YGNN CU VQ VJG EQORNGZKV[ CPF KPVGPUKV[ QH VJG CTQOC CPF VCUVG QH VJG HKPCN RTQFWEV (%QNG CPF òQDNG 1üüó; òQDNG 1üüû; öCODTGEJVU CPF 2TGVQTKWU ÿ000). 6JG EQORQPGPVU HQTOGF FWTKPI HGTOGPVCVKQP FQOKPCVG VJG XQNCVKNG RTQHKNG QH YKPGU, CU VJGUG EQORQWPFU CTG RTGUGPV KP VJG JKIJGUV EQPEGPVTCVKQPU (þQOG\ GV CN. 1üüû). 6JGTG KU C ENGCT PGGF HQT VJG FGXGNQROGPV QH YKPG [GCUVU VJCV EQWNF KORCTV URGEKHKE FGUKTCDNG EJCTCEVGTKUVKEU VQ C YKPG (þQOG\ GV CN. 1üüû; öCODTGEJVU CPF 2TGVQTKWU ÿ000). 4.4.4.1 Enhanced liberation of grape terpenoids ©P\[OCVKE J[FTQN[UKU QH VJG CTQOC RTGEWTUQTU RTGUGPV KP ITCRG LWKEG CU VJGKT TGURGEVKXG, PQP-XQNCVKNG O-IN[EQUKFGU (G.I., VGTRGP[N-IN[EQUKFGU) ECP NKDGTCVG VJG VGTRGPQNU (G.I., IGTCPKQN CPF PGTQN) VQ KPVGPUKH[ VJG XCTKGVCN EJCTCEVGT QH YKPGU (%CPCNöNCWDGTGU 1üü3). þTCRG-FGTKXGF IN[EQUKFCUGU CTG KPJKDKVGF D[ INWEQUG, GZJKDKV RQQT UVCDKNKV[ CV VJG NQY R* CPF JKIJ GVJCPQN NGXGNU QH YKPG, CPF CTG WPCDNG VQ J[FTQN[UG UWICT EQPLWICVGU QH VGTVKCT[ CNEQJQNU (G.I., NKPCNQQN). 6JGTGHQTG EQOOGTEKCN RTGRCTCVKQPU QH CTQOC-NKDGTCVKPI ¾-INWEQUKFCUGU CTG UQOGVKOGU CFFGF GKVJGT VQ HGTOGPVGF LWKEG CU UQQP CU VJG INWEQUG JCU DGGP EQPUWOGF D[ VJG [GCUV QT VQ [QWPI YKPG (%CPCN-öNCWDGTGU 1üü3). +V KU GZRGPUKXG, JQYGXGT, VQ CFF GZQIGPQWU GP\[OG RTGRCTCVKQPU VQ YKPG, CPF OCP[ VTCFKVKQPCNKUVU CPF RWTKUVU XKGY KV CU CP ÉCTVKHKEKCNÌ QT ÉWPPCVWTCNÌ KPVGTXGPVKQP D[ VJG YKPGOCMGT. 6JWU VJGTG KU KPVGTGUV KP HWPICN INWECPCUGU CPF [GCUV ¾INWEQUKFCUGU VJCV CTG PQV KPJKDKVGF D[ INWEQUG CPF VJCV EQWNF NKDGTCVG VJG ITCRG VGTRGPQNU FWTKPI HGTOGPVCVKQP (8CP 4GPUDWTI CPF 2TGVQTKWU ÿ000; öCODTGEJVU CPF 2TGVQTKWU ÿ000). 6JG CTQOC KPVGPUKV[ QH YKPG KPETGCUGF YJGP VJG ¾-1,û-INWECPCUG IGPG HTQO Trichoderma longibrachiatum YCU GZRTGUUGF KP YKPG [GCUV, RTGUWOCDN[ FWG VQ VJG J[FTQN[UKU QH IN[EQU[NCVGF HNCXQWT RTGEWTUQTU (2oTG\-þQP\hNG\ GV CN. 1üü3). öKMGYKUG, VJG S. cerevisiae GZQ-¾-1,3-INWECPCUG IGPG (EXG1) YCU QXGTGZRTGUUGF CPF VJG GPFQ-¾-1,û-INWECPCUG IGPG (END1) HTQO Butyrivibrio fibrisolvens, VJG GPFQ-¾-1,3-1,û-INWECPCUG IGPG (BEG1) HTQO Bacillus subtilis CPF VJG ½CTCDKPQHWTCPQUKFCUG IGPG (ABF2) (8CP 4GPUDWTI GV CN. 1üüù, 1üüø, 1üüõ) YGTG KPVTQFWEGF KPVQ S. cerevisiae. 5KPEG VJGUG ¾-INWEQUKFCUGU CTG CDUGPV HTQO OQUV S. cerevisiae UVCTVGT EWNVWTG UVTCKPU, VJG ¾-INWEQUKFCUG IGPG (BGL1) HTQO VJG [GCUV Saccharomycopsis fibuligera YCU GZRTGUUGF KP S. cerevisiae (8CP 4GPUDWTI GV CN. 1üüõ). 6TKCNU CTG WPFGTYC[ VQ FGVGTOKPG VJG GHHGEV QH VJGUG VTCPUIGPKE [GCUVU QP VJG XCTKGVCN EJCTCEVGT QH OWUECV YKPGU. 5QOG QH VJGUG CTQOC-NKDGTCVKPI GP\[OGU RTQXKFG CP CFFKVKQPCN CFXCPVCIG KP VJCV VJG[ CNUQ HCEKNKVCVG VJG TGOQXCN QH HKNVGTENQIIKPI RQN[UCEEJCTKFGU KP OWUV CPF YKPG (UGG 5GEVKQP û.ÿ.1).
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
1ÿø
4.4.4.2 Enhanced production of desirable volatile esters 6JG EJCTCEVGTKUVKE HTWKV[ QFQWTU QH YKPG CTG ECWUGF OCKPN[ D[ C OKZVWTG QH JGZ[N CEGVCVG, GVJ[N ECRTQCVG CPF ECRT[NCVG, KUQ-CO[N CEGVCVG CPF ÿ-RJGP[NGVJ[N CEGVCVG (öCODTGEJVU CPF 2TGVQTKWU ÿ000). +P S. cerevisiae, CNEQJQN CEGV[NVTCPUHGTCUG, GVJCPQN CEGV[NVTCPUHGTCUG CPF KUQ-CO[N CNEQJQN CEGV[NVTCPUHGTCUG CTG TGURQPUKDNG HQT VJG U[PVJGUKU QH CEGVCVG GUVGTU UWEJ CU KUQ-CO[N CEGVCVG CPF GVJ[N CEGVCVG (/CNEQTRU CPF &WHQWT 1üõø; /CNEQTRU GV CN. 1üü1). 6JGUG VJTGG UWNHJ[FT[N CEGV[NVTCPUHGTCUGU CTG GP\[OGU VJCV TGCEV YKVJ CEGV[N EQGP\[OG # (CEGV[N-%Q#) CPF, FGRGPFKPI QP VJG FGITGG QH CHHKPKV[, YKVJ XCTKQWU JKIJGT CNEQJQNU VQ RTQFWEG GUVGTU (÷WLKK GV CN. 1üüû, 1üüù, 1üüø). 6JG DGUV UVWFKGF QH VJGUG CEGV[NVTCPUHGTCUGU, VJG ATF1-GPEQFGF CNEQJQN CEGV[NVTCPUHGTCUG CEVKXKV[, KU NQECVGF YKVJKP VJG [GCUV•U EGNNWNCT XCEWQNGU CPF RNC[U C EGPVTCN TQNG KP VJG RTQFWEVKQP QH KUQ-CO[N CEGVCVG CPF, VQ C NGUUGT GZVGPV, GVJ[N CEGVCVG FWTKPI HGTOGPVCVKQP. 9JGP VJG ATF1 IGPG YCU QXGTGZRTGUUGF WPFGT VJG EQPVTQN QH VJG EQPUVKVWVKXG PGK1 RTQOQVGT, VJG VTCPUHQTOGF YKPG [GCUV UVTCKPU RTQFWEGF YKPG CPF FKUVKNNCVGU YKVJ UKIPKHKECPVN[ GNGXCVGF NGXGNU QH GVJ[N CEGVCVG, KUQ-CO[N CEGVCVG CPF ÿ-RJGP[NGVJ[N CEGVCVG (öKNN[ GV CN. ÿ000). 6JGTG YGTG QPN[ OKPQT EJCPIGU KP VJG EQPEGPVTCVKQP QH GVJ[N ECRTCVG, GVJ[N ECRT[NCVG CPF JGZ[N CEGVCVG, YJGTGCU VJG CEGVKE CEKF EQPEGPVTCVKQP FGETGCUGF D[ OQTG VJCP JCNH. 6JGUG EJCPIGU JCF C OCTMGF GHHGEV QP VJG FGUKTCDNG HTWKV[ñHNQYGT[ CTQOC CPF VJG UQNXGPVñEJGOKECN EJCTCEVGT QH %JGPKP DNCPE YKPGU CPF QP VJG UQNXGPVñEJGOKECN, JGTDCEGQWU CPF JGCFU-CUUQEKCVGF CTQOC QH DTCPF[ (öKNN[ GV CN. ÿ000). 6JKU UVWF[ GUVCDNKUJGF VJCV VJG QXGTGZRTGUUKQP QH CEGV[NVTCPUHGTCUG IGPGU, UWEJ CU ATF1, EQWNF JCXG C RTQHQWPF GHHGEV QP VJG HNCXQWT RTQHKNGU QH YKPGU CPF FKUVKNNCVGU FGHKEKGPV KP CTQOC CPF QP VJG TGVGPVKQP QH C HTWKVKGT EJCTCEVGT HQT NQPIGT RGTKQFU CHVGT DQVVNKPI. 4.4.4.3 Optimised production of glycerol and higher alcohols þN[EGTQN KU DGNKGXGF VQ KORCTV C UNKIJVN[ UYGGV VCUVG VQ YKPG CPF VQ EQPVTKDWVG VQ UOQQVJPGUU, EQPUKUVGPE[, QXGTCNN DQF[ CPF OQWVJ HGGN (2TKQT CPF *QJOCPP 1üüø; 5ECPGU GV CN. 1üüõ). 9KPG [GCUV UVTCKPU VJCV QXGTRTQFWEG IN[EGTQN VJGTGHQTG EQWNF DG XCNWCDNG KP KORTQXKPI QTICPQNGRVKE SWCNKV[ (/KEJPKEM GV CN. 1üüø; 4GOK\G GV CN. 1üüü). 5GXGTCN NCDQTCVQTKGU JCXG TGRQTVGF VJCV VJG QXGTGZRTGUUKQP QH GKVJGT QH VJG S. cerevisiae GPD1- CPF GPD2-GPEQFGF IN[EGTQN-3-RJQURJCVG FGJ[FTQIGPCUG KUQ\[OGU TGUWNVU KP YKPGU YKVJ UKIPKHKECPVN[ JKIJGT NGXGNU QH IN[EGTQN (/KEJPKEM GV CN. 1üüø; 4GOK\G GV CN. 1üüü; &G $CTTQU öQRGU GV CN. ÿ000). 5KFG GHHGEVU QH VJGUG IN[EGTQN-QXGTRTQFWEKPI [GCUVU KPENWFG NQYGT DKQOCUU RTQFWEVKQP CPF VJG CEEWOWNCVKQP QH D[RTQFWEVU UWEJ CU R[TWXCVG, CEGVCVG, CEGVQKP, ÿ,3-DWVCPGFKQN CPF UWEEKPCVG (/KEJPKEM GV CN. 1üüø; 4GOK\G GV CN. 1üüü; &G $CTTQU öQRGU GV CN. ÿ000; ©INKPVQP GV CN. ÿ00ÿ). 6Q QXGTEQOG C OCTMGF KPETGCUG KP CEGVCVG HQTOCVKQP, VJG OQUV PGICVKXG UKFG GHHGEV QH IN[EGTQN QXGTRTQFWEVKQP, VJG ALD6 IGPG, GPEQFKPI C E[VQUQNKE /Iÿë-CEVKXCVGF, ò#&2-FGRGPFGPV CEGVCNFGJ[FG FGJ[FTQIGPCUG, YCU FKUTWRVGF. # YKPG [GCUV UVTCKP KP YJKEJ GPD1 QT GPD2 YCU QXGTGZRTGUUGF KP EQPLWPEVKQP YKVJ VJG FGNGVKQP QH ALD6 RTQFWEGF VYQ- VQ VJTGG-HQNF OQTG IN[EGTQN CPF C UKOKNCT COQWPV QH CEGVCVG EQORCTGF VQ VJG WPVTCPUHQTOGF UVTCKP (4GOK\G GV CN.
1ÿõ
+UCM 5. 2TGVQTKWU
1üüü; ©INKPVQP GV CN. ÿ00ÿ). 6Q OCKPVCKP VJG TGFQZ DCNCPEG KP VJGUG TGEQODKPCPV YKPG [GCUVU, VJG HQTOCVKQP QH UWEEKPCVG CPF ÿ,3-DWVCPGFKQN YCU KPETGCUGF VQ EQPEGPVTCVKQPU TGOCKPKPI KP VJG TCPIG QH VJCV HQWPF KP YKPG. #P CPCN[UKU QH VJG OGVCDQNQOG QH VJGUG UVTCKPU UJQYGF VJCV VJG RTQHKNG QH UQOG UGEQPFCT[ OGVCDQNKVGU QH HGTOGPVCVKQP, KPENWFKPI CEKFU, GUVGTU, CNFGJ[FGU CPF JKIJGT CNEQJQNU, OCP[ QH YJKEJ CTG HNCXQWT-CEVKXG KP YKPG, YCU FKHHGTGPV VQ VJCV QH VJG EQPVTQN YKPG (©INKPVQP GV CN. ÿ00ÿ). 6JWU, OQFKH[KPI VJG GZRTGUUKQP QH GPD1 QT GPD2, VQIGVJGT YKVJ ALD6, TGRTGUGPVU CP GHHGEVKXG UVTCVGI[ VQ KPETGCUG VJG IN[EGTQN EQPEGPVTCVKQP CPF FGETGCUG VJG GVJCPQN EQPEGPVTCVKQP FWTKPI HGTOGPVCVKQP, CPF CNVGTU VJG EJGOKECN EQORQUKVKQP QH YKPG KP UWEJ C YC[ VJCV PQXGN HNCXQWT FKXGTUKV[ KU C RQUUKDKNKV[ (©INKPVQP GV CN. ÿ00ÿ). 7PNKMG IN[EGTQN, [GCUV-FGTKXGF HWUGN QKNU (JKIJGT CNEQJQNU UWEJ CU KUQDWV[N, KUQCO[N CPF CEVKXG CO[N CNEQJQN) JCXG WPFGUKTCDNG HNCXQWT CPF QFQWT EJCTCEVGTKUVKEU, GURGEKCNN[ KH VJG[ JCXG DGGP EQPEGPVTCVGF VQ C JKIJ FGITGG KP FKUVKNNGF RTQFWEVU (5PQY 1üõ3; þKWFKEK GV CN. 1üü0). 6JG WUG QH +NG-, öGW- CPF 8CN- CWZQVTQRJKE OWVCPVU ECP JCNXG VJG NGXGNU QH KUQDWVCPQN, CEVKXG CO[N CNEQJQN CPF KUQCO[N CNEQJQN RTQFWEGF KP HGTOGPVCVKQPU, DWV DGECWUG VJGKT ITQYVJ CPF HGTOGPVCVKQP TCVGU CTG EQORTQOKUGF, VJGUG OWVCPVU CTG QH PQ EQOOGTEKCN WUG (+PITCJCO CPF þW[OQP 1üù0; +PITCJCO GV CN. 1üù1; 5PQY 1üõ3). 6Q FCVG, PQ CNVGTPCVKXG CRRTQCEJ VQ QXGTEQOG VJKU FKHHKEWNV[ JCU DGGP TGRQTVGF. 4.4.4.4 Bio-adjustment of wine acidity 6JG CEKFKV[ QH ITCRG LWKEG CPF YKPG, YJKEJ KU OCKPN[ CVVTKDWVGF VQ VCTVCTKE CPF OCNKE CEKF, JCU C RTQHQWPF KPHNWGPEG QP UGPUQT[ SWCNKV[. #EKFKV[ CNUQ CHHGEVU YKPG•U RJ[UKECN UVCDKNKV[ (G.I., VJG UQNWDKNKV[ QH RTQVGKPU CPF VCTVTCVG UCNVU, CPF VJG GHHGEVKXGPGUU QH DGPVQPKVG VTGCVOGPV CU FGUETKDGF RTGXKQWUN[), DKQEJGOKECN UVCDKNKV[ (G.I., VJG GHHGEVKXGPGUU QH CPVKQZKFCPVU CPF GP\[OG CFFKVKQPU; VJG RQN[OGTKUCVKQP QH EQNQWT RKIOGPVU; CPF VJG QZKFCVKXG CPF DTQYPKPI TGCEVKQPU) CPF OKETQDKCN UVCDKNKV[ (G.I., VJG GHHGEVKXGPGUU QH CPVKOKETQDKCN EQORQWPFU CU YGNN CU VJG UWTXKXCN CPF ITQYVJ QH CNN YKPG OKETQDGU) (%CRWVK CPF 4[CP 1üüù). +P EQQNGT YKPG RTQFWEKPI TGIKQPU, VJG VKVTCVCDNG CEKFKV[ QH ITCRG LWKEG CPF YKPG OWUV DG TGFWEGF D[, HQT GZCORNG, DNGPFKPI QT EJGOKECN PGWVTCNKUCVKQP D[ VJG CFFKVKQP QH ECNEKWO ECTDQPCVG CPF RTGEKRKVCVKQP, YJGTGCU KP YCTOGT XKVKEWNVWTCN TGIKQPU, VJG CFLWUVOGPV QH YKPG CEKFKV[ IGPGTCNN[ GPVCKNU KPETGCUKPI VJG VKVTCVCDNG CEKFKV[ (G.I., CFFKPI VCTVCTKE, OCNKE QT EKVTKE CEKFU). 6JGUG RTCEVKEGU TGSWKTG GZVGPUKXG NCDQWT CPF ECRKVCN KPRWV CPF ECP TGFWEG YKPG SWCNKV[. $KQFGCEKFKHKECVKQP QH YKPG KU EWTTGPVN[ OGFKCVGF D[ NCEVKE CEKF DCEVGTKC (KP RCTVKEWNCT Oenococcus oeni), YJKEJ FGECTDQZ[NCVG OCNKE CEKF VQ NCEVKE CEKF CPF ECTDQP FKQZKFG FWTKPI OCNQNCEVKE HGTOGPVCVKQP. *QYGXGT, UVWEM QT UNWIIKUJ OCNQNCEVKE HGTOGPVCVKQP QHVGP QEEWTU DGECWUG QH PWVTKGPV NKOKVCVKQP, NQY VGORGTCVWTG, CEKFKE R*, CPF JKIJ CNEQJQN CPF UWNRJWT FKQZKFG NGXGNU KP YKPG. 5GXGTCN CNVGTPCVKXGU JCXG DGGP GZRNQTGF, KPENWFKPI VJG WUG QH OCNCVG-FGITCFKPI [GCUVU. /CNQGVJCPQNKE HGTOGPVCVKQPU EQPFWEVGF D[ S. pombe ECWUGF OCNCVG VQ DG EQPXGTVGF GHHGEVKXGN[ VQ GVJCPQN, DWV QHH-HNCXQWTU YGTG RTQFWEGF (þCNNCPFGT 1üøø). 7PNKMG S. pombe, S. cerevisiae NCEMU CP CEVKXG OCNCVG VTCPURQTV U[UVGO, CPF VJG UWDUVTCVG
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
1ÿü
URGEKHKEKV[ QH KVU OCNKE GP\[OG KU CDQWV 1ó-HQNF NQYGT VJCP VJCV QH VJG S. pombe OCNKE GP\[OG (8QNUEJGPM GV CN. 1üüø). 6Q GPIKPGGT C OCNQ-GVJCPQNKE YKPG [GCUV, VJG mae1 OCNCVG RGTOGCUG IGPG CPF VJG mae2 OCNKE GP\[OG IGPG HTQO S. pombe YGTG EQ-GZRTGUUGF KP S. cerevisiae (8QNUEJGPM GV CN. 1üüø). 5KOKNCTN[, KP CP CVVGORV VQ GPIKPGGT C OCNQNCEVKE RCVJYC[ KP S. cerevisiae, VJG OCNQNCEVKE IGPGU (mle5) HTQO Lactococcus lactis YGTG EQ-GZRTGUUGF YKVJ VJG S. pombe mae1 RGTOGCUG IGPG (8QNUEJGPM GV CN. 1üüø). &WTKPI XKPKHKECVKQP VTKCNU, KV YCU UJQYP VJCV VJGUG OCNQ-GVJCPQNKE CPF OCNQNCEVKE YKPG [GCUVU YGTG CDNG VQ FGITCFG CNN VJG OCNKE CEKF KP OWUV YKVJKP VJTGG FC[U CPF PQ QHH-HNCXQWT YCU RTQFWEGF KP VJG YKPG. +P EQPVTCUV, VJG DKQ-CEKFKHKECVKQP QH JKIJ R* YKPGU ECP DG HCEKNKVCVGF D[ C UVTCKP QH S. cerevisiae KP YJKEJ VJG NCEVKEQFGJ[FTQIGPCUG-GPEQFKPI IGPG HTQO Lactobacillus casei (&GSWKP CPF $CTTG 1üüû) CPF VJG DQXKPG NCEVKEQFGJ[FTQIGPCUGGPEQFKPI IGPG (2QTTQ GV CN. 1üüó) YGTG GZRTGUUGF. 9JGP VJG L. casei NCEVKEQFGJ[FTQIGPCUG IGPG YCU GZRTGUUGF WPFGT VJG EQPVTQN QH VJG [GCUV CNEQJQN FGJ[FTQIGPCUG + (ADH1) RTQOQVGT KP YKPG [GCUVU, WR VQ ÿ0ì QH VJG INWEQUG YCU EQPXGTVGF KPVQ ö-NCEVKE CEKF, DWV VJG HGTOGPVCVKQP TCVG YCU UNQYGT (&GSWKP GV CN. 1üüü). 4.4.4.5 Elimination of phenolic off-flavour ÷TGG RJGPQNKE CEKFU, UWEJ CU Ð-EQWOCTKE CPF HGTWNKE CEKFU, ECP DG OGVCDQNKUGF KPVQ û-XKP[N CPF û-GVJ[N FGTKXCVGU D[ XCTKQWU OKETQQTICPKUOU KP YKPG. 6JGUG XQNCVKNG RJGPQNU EQPVTKDWVG VQ VJG CTQOC QH VJG YKPG, DWV, KH VJG[ GZEGGF EGTVCKP VJTGUJQNF XCNWGU, VJG[ ECP KORCTV C OGFKEKPCN HNCXQWT. S. cerevisiae EQPVCKPU VJG PAD1 IGPG (CNUQ MPQYP CU POF1), YJKEJ GPEQFGU C RJGP[N CET[NKE CEKF FGECTDQZ[NCUG YKVJ NQY CEVKXKV[. +P CP CVVGORV VQ QRVKOKUG VJG NGXGNU QH RJGPQNKE CEKFU KP YKPG, [GCUV UVTCKPU YGTG EQPUVTWEVGF VJCV ECTTKGF GKVJGT C FKUTWRVGF CNNGNG QH VJG S. cerevisiae PAD1 IGPG, QT ECTTKGF EQPUVKVWVKXGN[ GZRTGUUGF EQRKGU QH PAD1, CU YGNN CU VJG Bacillus subtilis RJGPQNKE CEKF FGECTDQZ[NCUG IGPG (padc) CPF VJG Lactobacillus plantarum Ð-EQWOCTKE CEKF FGECTDQZ[NCUG IGPG (pdc) (5OKV GV CN. ÿ003). 6JG QXGTGZRTGUUKQP QH VJG VYQ TGEQODKPCPV DCEVGTKCN IGPGU, pdc CPF padc, UJQYGF JKIJ GP\[OG CEVKXKV[ KP S. cerevisiae, YJGTGCU VJG QXGTGZRTGUUGF PAD1 FKF PQV TGUWNV KP CP[ JKIJGT RJGP[N CET[NKE CEKF FGECTDQZ[NCUG CEVKXKV[ VJCP VJG EQPVTQN UVTCKP. 9KPG OCFG YKVJ [GCUVU KP YJKEJ VJG PAD1 IGPG YCU FKUTWRVGF CPF KP YJKEJ VJG B. subtilis pdc CPF padc IGPGU YGTG QXGTGZRTGUUGF EQPVCKPGF JKIJGT NGXGNU QH XQNCVKNG RJGPQNU. +V YCU CNUQ HQWPF VJCV VJG QXGTGZRTGUUKQP QH VJG padc IGPG KP YKPG [GCUVU TGUWNVGF KP YKPG YKVJ GNGXCVGF NGXGNU QH HCXQWTCDNG OQPQVGTRGPGU. 6JKU UVWF[ OKIJV NGCF VQ VJG FGXGNQROGPV QH YKPG [GCUV UVCTVGT EWNVWTGU HQT RTQFWEKPI QRVKOCN NGXGNU QH XQNCVKNG RJGPQNU, VJGTGD[ KORTQXKPI VJG UGPUQTKCN SWCNKV[ QH YKPG. 4.4.4.6 Reduced sulphite and sulphide production *[FTQIGP UWNRJKFG (*ÿ5), HQTOGF HTQO UWNRJCVG, UWNRJKVG, GNGOGPVCN UWNRJWT (CRRNKGF CU C HWPIKEKFG) CPF E[UVGKPG, KU QPG QH VJG OQUV WPFGUKTCDNG QH [GCUV OGVCDQNKVGU, UKPEG KV ECWUGU CP QHH-HNCXQWT TGOKPKUEGPV QH TQVVGP GIIU (5PQY 1üõ3; 4CWJWV 1üü3). 5GXGTCN HCEVQTU CHHGEV UWNRJKVG CPF UWNRJKFG HQTOCVKQP, KPENWFKPI VJG IGPGVKE DCEMITQWPF QH VJG [GCUV UVTCKP KPXQNXGF, VJG PWVTKGPV EQORQUKVKQP QH VJG ITCRG
130
+UCM 5. 2TGVQTKWU
LWKEG, VJG NGXGN QH 5ýÿ CFFGF VQ VJG OWUV UJQTVN[ DGHQTG KPQEWNCVKPI YKVJ [GCUV, VJG EQPEGPVTCVKQP QH UWNRJCVG, OWUV ENCTKHKECVKQP, CPF VJG KPKVKCN R* CPF VGORGTCVWTG (4CWJWV 1üü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ÿ00ÿ). ÷KPCNN[, UWNRJKVG KU TGFWEGF VQ UWNRJKFG D[ UWNRJKVG TGFWEVCUG, YJKEJ KU GPEQFGF D[ MET10 CPF MET5 (HQT OQTG FGVCKNU CDQWV VJG /©6 RCVJYC[, RNGCUG UGG %JCRVGT ó). +V CRRGCTU VJCV VJG OQUV GHHGEVKXG YC[ VQ FGXGNQR YKPG [GCUV UVTCKPU YKVJ C FGETGCUGF CDKNKV[ VQ RTQFWEG UWNRJKVG CPF UWNRJKFG KU VQ FKUTWRV VJG MET14 CFGPQU[NRJQURJQUWNRJCVG MKPCUG IGPG QT VJG MRX1 OGVJKQPKPG UWNRJQZKFG TGFWEVCUG IGPG (&QPCNKGU CPF 5VCJN ÿ00ÿ).
4.ÿ %QPENWUKQPU CPF HWVWTG RGTURGEVKXGU #P KPETGCUKPI ICR DGVYGGP YKPG RTQFWEVKQP CPF YKPG EQPUWORVKQP, UJKHVKPI EQPUWOGT RTGHGTGPEGU, INQDCNKUCVKQP CPF QVJGT GEQPQOKE HCEVQTU JCXG HQTEGF CP GXQNWVKQP QH VJG KPVGTPCVKQPCN YKPG KPFWUVT[ HTQO C RTQFWEVKQP-FTKXGP "EQVVCIG KPFWUVT[" VQ INQDCN PGVYQTMU QH EQPUWOGT-EQPUEKQWU RTQFWEGTU ($KUUQP GV CN. ÿ00ÿ). ýPG TGUWNV KU CP WTIGPV ECNN HQT VJG HWTVJGT GPJCPEGOGPV QH YKPG SWCNKV[, RWTKV[, WPKSWGPGUU CPF FKXGTUKV[ (2TGVQTKWU CPF $CWGT ÿ00ÿ). +V KU ENGCTN[ FGUKTCDNG, VQ RWV KV OKNFN[, VJCV VJG KPVGTPCVKQPCN YKPG EQOOWPKV[ TGURQPF KP C VKOGN[ HCUJKQP VQ VJG EJCNNGPIGU CPF QRRQTVWPKVKGU CUUQEKCVGF YKVJ VJG VTCPUKVKQP VQ C OCTMGV-QTKGPVGF KPFWUVT[. /CP[, VJQWIJ PQV CNN, YKPG-RTQFWEKPI EQWPVTKGU JCXG GPFQTUGF VJG RTKPEKRNG VJCV VJKU VTCPUKVKQP KU C OCVVGT QH JKIJ UVTCVGIKE KORQTVCPEG. 6JKU EJCRVGT RTQRQUGU VJCV IGPG VGEJPQNQI[ JCU C ITGCV FGCN VQ QHHGT VJG YKPG KPFWUVT[ KP KVU UGCTEJ HQT YC[U VQ OGGV EQPUWOGT FGOCPF HQT VJG DNGPFKPI QH SWCNKV[, PWVTKVKQPCN DGPGHKVU CPF UQWPF GPXKTQPOGPVCN RTCEVKEGU. #V RTGUGPV VJGTG KU C PQVGF NCEM QH EQPUGPUWU CU VQ YJGVJGT IGPG VGEJPQNQI[ UJQWNF DG XKGYGF CU EGPVTCN QT RGTKRJGTCN VQ VJG HWVWTG QH VJG KPVGTPCVKQPCN YKPG KPFWUVT[. $WV C ITQYKPI UGIOGPV QH VJG YKPG EQOOWPKV[ GZRGEVU VJCV GZRCPFKPI MPQYNGFIG KP UGPUQ-EJGOKUVT[, HWPFCOGPVCN OQNGEWNCT IGPGVKEU CPF DKQKPHQTOCVKEGU (KPENWFKPI IGPQOKEU, VTCPUETKRVQOKEU, RTQVGQOKEU CPF OGVCDQNQOKEU) YKNN CEEGNGTCVG VJG RCEG CV YJKEJ EWUVQOKUGF YKPG [GCUVU CTG FGXGNQRGF HQT C OCTMGVFTKXGP CPF SWCNKV[-HQEWUGF KPFWUVT[ ($KUUQP GV CN. ÿ00ÿ; 2TGVQTKWU CPF $CWGT, ÿ00ÿ; 8KXKGT CPF 2TGVQTKWU ÿ00ÿ). +PFWUVT[ NGCFGTU DGNKGXG VJCV YKPG TGUGCTEJ ECP PQ NQPIGT HWPEVKQP YKVJKP VJG NKOKVCVKQPU QH C EQPXGPVKQPCN, WPKFKOGPUKQPCN EQPVKPWWO, QP YJKEJ DCUKE CPF CRRNKGF TGUGCTEJ ECPPQV DG ENQUGT VQ QPG QH VJG RQNGU QH VJG EQPVKPWWO YKVJQWV DGKPI
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
131
(KI. û. # UEJGOCVKE TGRTGUGPVCVKQP QH VJG WPKFKOGPUKQPCN (#) CPF SWCFTCPV ($) OQFGNU HQT ENCUUKH[KPI TGUGCTEJ. 6JG EQPXGPVKQPCN RCTCFKIO KP YJKEJ UEKGPEG KU ENCUUKHKGF QP C WPKFKOGPUKQPCN DCUKEñCRRNKGF EQPVKPWWO KU QWVFCVGF. 7UKPI VJG SWCFTCPV OQFGN (5VQMGU 1üüø), 2CUVGWT•U SWCFTCPV KU WPFGT-TGRTGUGPVGF KP VJG HKGNF QH YKPG [GCUV TGUGCTEJ.
HWTVJGT CYC[ HTQO VJG QVJGT. 9KPG TGUGCTEJ UJQWNF TCVJGT DG ENCUUKHKGF CPF EQPFWEVGF CEEQTFKPI VQ 5VQMGU• ÉSWCFTCPVÌ OQFGN, YJKEJ KPXQNXGU KPVGITCVGF TCVJGT VJCP UGSWGPVKCN RWTUWKV QH C HWPFCOGPVCN WPFGTUVCPFKPI (5VQMGU 1üüø). É2CUVGWT•U SWCFTCPVÌ (÷KI. ù) UJQYU ITCRJKECNN[ VJCV YKPG TGUGCTEJ PGGF PQV DG GKVJGT HWPFCOGPVCN QT CRRNKGF, DWV OC[ DG DQVJ UKOWNVCPGQWUN[. öGCFKPI QGPQNQIKUVU HGGN VJCV VJG DGPGHKVU CXCKNCDNG HTQO VJG VTKCN-CPF-GTTQT É©FKUQPKCPÌ CRRTQCEJ (÷KI. ù)
13ÿ
+UCM 5. 2TGVQTKWU
6CDNG 4. %JCNNGPIGU HCEKPI VJG EQOOGTEKCNKUCVKQP QH TGEQODKPCPV YKPG [GCUVU úDUVCENGU $TKGH FGUETKRVKQP QH VJG KUUWGU CV UVCMG CPF EJCNNGPIGU 5EKGPVKHKE 8“ %QORNGZKV[ QH YKPG OCMGU KFGPVKHKECVKQP QH IGPGVKE PGGFU FKHHKEWNV. CPF VGEJ- 8“ 6JG TGSWKTGOGPVU QH RTQFWEGTU, EQPUWOGTU CPF TGIWNCVQT[ CWVJQTKVKGU OC[ EQPHNKEV. PKECN JWT- 8“ 6GEJPQNQI[ HQT NCDQTCVQT[ UVTCKPU KU PQV [GV UWHHKEKGPV HQT EQOOGTEKCN UVTCKPU. *QYGXGT, PGY VGEJPQNQI[ KU OCMKPI RQUUKDNG VJG CPCN[UKU QH GPVKTG IGPQOGU, VTCPUETKRFNGU
VQOGU, RTQVGQOGU CPF OGVCDQNQOGU. þ/ ÉRTQVQV[RGÌ YKPG [GCUVU JCXG DTQWIJV UGXGTCN VCTIGVU QH VJG YKPG EQOOWPKV[ YKVJKP VJG TGCNOU QH RQUUKDKNKV[. þWCTCPVGGU TGSWKTGF HQT VJG CRRTQXCN QH þ/ RTQFWEVU CPF VJG TGNGCUG QH þ/ýU: 8“ C EQORNGVG FGHKPKVKQP QH VJG &ò# UGSWGPEG. 8“ VJG GNKOKPCVKQP QH CP[ UGSWGPEG FKURGPUCDNG HQT GZRTGUUKQP QH VJG FGUKTGF RTQRGTV[. 8“ VJG CDUGPEG QH CP[ UGNGEVKXG CFXCPVCIG EQPHGTTGF QP VJG VTCPUIGPKE QTICPKUO VJCV EQWNF CNNQY KV VQ DGEQOG FQOKPCPV KP PCVWTCN JCDKVCVU. 8“ PQ FCPIGT VQ JWOCP JGCNVJ CPFñQT VJG GPXKTQPOGPV HTQO VJG VTCPUHQTOGF &ò#. 8“ C ENGCT CFXCPVCIG VQ DQVJ VJG RTQFWEGT CPF VJG EQPUWOGT. 6JG EQPEGRV QH ÉUWDUVCPVKCN GSWKXCNGPEGÌ GPVCKNU VJCV: 8“ YJGP UWDUVCPVKCN GSWKXCNGPEG ECP DG FGOQPUVTCVGF, PQ HWTVJGT UCHGV[ EQPUKFGTCVKQPU CTG WUWCNN[ PGEGUUCT[ 8“ YJGP UWDUVCPVKCN GSWKXCNGPEG KU PQV EQPXKPEKPIN[ UJQYP, VJG RQKPVU QH FKHHGTGPEG OWUV DG UWDLGEVGF VQ HWTVJGT UCHGV[ UETWVKP[. 8“ 2CVGPVU EQXGTKPI OCP[ IGPGVKE VQQNU CPF OGVJQFU TGUVTKEV ÉHTGGFQO VQ QRGTCVGÌ. 8“ ÷QTOCN CITGGOGPVU CTG PGGFGF VQ CFFTGUU KPVGNNGEVWCN RTQRGTV[ KUUWGU UWEJ CU RCVGPVU QT QVJGT HQTOU QH RTQVGEVKQP QH IGPGU, RTQOQVGTU CPF VGEJPQNQIKGU. 8“ 6JG TGNGCUG QH þ/ YKPG [GCUV UVTCKPU YKVJ FKURWVGF QYPGTUJKR EQWNF DG FGVTKOGPVCN VQ EQOOGTEKCNKUCVKQP. 8“ &GXGNQRGTU QH IGPGVKECNN[ KORTQXGF YKPG [GCUVU YKVJ ÉUWHHKEKGPVN[ FKUVKPEVÌ RTQRGTVKGU OWUV CNUQ DG RTQVGEVGF. 8“ ýTICPKUGF RQNKVKECN CEVKXKV[ FKTGEVGF CV þ/ýU. 8“ %NCKOU VJCV þ/ý RCVGPVU IKXG WPHCKT CFXCPVCIG CPF EQPEGPVTCVG GEQPQOKE RQYGT. 8“ 2QNKVKECN KPVGTHGTGPEG HTQO VJQUG YKVJ XGUVGF KPVGTGUVU KP GEQPQOKE CPF CITKEWNVWTCN RTQVGEVKQPKUO. 8“ &GNKDGTCVG OKUWUG QH UEKGPVKHKE FCVC VQ LWUVKH[ VTCFG DCPU CPF VGEJPKECN DCTTKGTU VQ HTGG VTCFG. 8“ %CNNU HQT URGEKHKE NCDGNNKPI KPETGCUG VJG YKPG KPFWUVT[•U JGUKVCPEG VQ CFQRV þ/ YKPG [GCUVU. 8“ 4KUM QH ÉDCEMNCUJÌ OCTMGV YKVJ NCDGNU UVCVKPI VJCV C RCTVKEWNCT YKPG RTQFWEV KU Éþ/ HTGG.Ì
8“
öGICN CPF TGIWNCVQT[ JWTFNGU
+PVGNNGEVWCN RTQRGTV[ CPF RCVGPVKPI JWTFNGU 2QNKVKECN CPF GEQPQOKE JWTFNGU 6TCFKVKQPCN CPF EWNVWTCN JWTFNGU 2WDNKE RGTEGRVKQP JWTFNGU
÷GCT-OQPIGTKPI URTGCFU OQTG TGCFKN[ VJCP IQQF UGPUG QT YKUG UEKGPEG, CPF C UKIPKHKECPV RTQRQTVKQP QH VJG RWDNKE UVKNN UWURGEVU VJCV: 8“ þ/ HQQF YKNN RTQXG WPJGCNVJ[ KP VJG NQPI VGTO 8“ GUECRG QH þ/ýU YKNN FCOCIG VJG GPXKTQPOGPV CPF TGUWNV KP C NQUU QH DKQFKXGTUKV[ 8“ VJGTG KU KPUWHHKEKGPV NGICN CPF RTCEVKECN RTQVGEVKQP CICKPUV CEEKFGPVU KPXQNXKPI þ/ýU. 6JG EQPUWOGT UJQWNF DG 8“ TGCUUWTGF QH HKTUV-ENCUU, VTCPURCTGPV TGIWNCVQT[ U[UVGOU CPF VJG OGVKEWNQWU KORNGOGPVCVKQP QH DKQUCHGV[ NGIKUNCVKQP YKVJ ENGCT VGEJPKECN UVCPFCTFU CPF FGHKPKVKQPU YKVJ TGURGEV VQ þ/ RTQFWEVU 8“ RGTUWCFGF D[ RTQRGT TKUM CUUGUUOGPV CPF ENGCT FGOQPUVTCVKQP QH UCHGV[, CPF VJWU DG GORQYGTGF VQ OCMG KPHQTOGF FGEKUKQPU 8“ CUUWTGF VJCV þ/ YKPG CPF QVJGT ITCRG-FGTKXGF RTQFWEVU YKNN PQV DG ÉHQTEG-HGFÌ FQYP VJGKT VJTQCVU HQT RTQHKV YJGP VJGTG KU PQ ENGCT CFXCPVCIG HQT VJGO.
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
133
JCXG DGGP NCTIGN[ GZJCWUVGF CPF VJCV HWVWTG CFXCPEGU CTG DGUV RWTUWGF KP VJG EQPVGZV QH CP KPENWUKXG É2CUVGWTKCPÌ CRRTQCEJ. #EEQTFKPI VQ VJGUG GZRGTVU YJCV KU OQUV PGGFGF KU PQV DCUKE TGUGCTEJ (É$QJT•U SWCFTCPVÌ KP ÷KIWTG ù) KP VJG VGTO•U EWTTGPV WUCIG. 9JCV KU PGGFGF, TCVJGT, KU YKPG TGUGCTEJ FKTGEVGF VQYCTF KPETGCUKPI HWPFCOGPVCN WPFGTUVCPFKPI KP C EQPVGZV TGURQPUKXG VQ VJG CRRNKGF PGGFU QH YKPG RTQFWEGTU CPF EQPUWOGTU CV NGXGNU QH DQVJ RTQDNGO UGNGEVKQP CPF GZRGTKOGPVCN FGUKIP. #FXCPEGU KP UWEJ ÉCRRNKGF HWPFCOGPVCNUÌ RTQXKFG WPFGTUVCPFKPI VJCV GPCDNGU KPPQXCVKQP. 6JGTGHQTG, YKPG [GCUV TGUGCTEJ KPURKTGF D[ DQVJ VJG SWGUV HQT WPFGTUVCPFKPI VJG DCUKE OQNGEWNCT DKQNQI[ QH [GCUVU CPF D[ EQPUKFGTCVKQPU QH KVU WUG, RTQOKUG VQ DG C RQYGTHWN F[PCOQ QH VGEJPQNQIKECN RTQITGUU KP VJG KPETGCUKPIN[ OCTMGV-QTKGPVCVGF INQDCN YKPG KPFWUVT[. 4GCNKUKPI VJG RQVGPVKCN DGPGHKVU QH CRRN[KPI IGPG VGEJPQNQI[ KP VJG YKPG KPFWUVT[ KU D[ PQ OGCPU EGTVCKP. 6JGTG TGOCKP C OWNVKVWFG QH EQORNGZ, KPVGTEQPPGEVGF UEKGPVKHKE, VGEJPKECN, GEQPQOKE, OCTMGVKPI, UCHGV[, NGICN CPF GVJKECN DCTTKGTU. òQ TGEQODKPCPV YKPG [GCUV JCU DGGP WUGF VQ FCVG QP C EQOOGTEKCN UECNG, CPF YQP•V DG WPVKN DQVJ EQPUWOGTU CPF KPFWUVT[ CTG UCVKUHKGF VJCV þ/ RTQFWEVU CTG UCHG, QH JKIJ SWCNKV[ CPF DGPGHKEKCN (6CDNG û). þKXGP VJG EWTTGPV FGGRN[ TQQVGF EQPEGTPU QH EQPUWOGTU CPF VTCFKVKQPCNKUVU, KV EQWNF DG C EQOOGTEKCN FKUCUVGT VQ OCTMGV VJG HKTUV YKPG HGTOGPVGF YKVJ TGEQODKPCPV [GCUV. *QYGXGT, KV KU CNUQ ENGCT VJCV KV YQWNF DG GSWCNN[ FKUCUVTQWU VQ KIPQTG IGPG VGEJPQNQI[, YKVJ KVU RJGPQOGPCN RQVGPVKCN VQ LGVRTQRGN VJG YKPG KPFWUVT[ KPVQ VJG KPGXKVCDNG GTC QH Édesigner” RTQFWEVU. 6JG CRRNKECVKQP QH IGPG VGEJPQNQI[ QHHGTU XCUV RQVGPVKCN DGPGHKV VQ VJG YKPG EQPUWOGT CPF RTQFWEGT CNKMG, KH VJG CRRNKECVKQP KU LWFKEKQWU, U[UVGOCVKE CPF FQPG YKVJ JKIJ TGICTF HQT VJG WPKSWG PCVWTG QH VJG RTQFWEV. 6JG HKTUV þ/ YKPG RTQFWEVU UJQWNF WPGSWKXQECNN[ FGOQPUVTCVG OKPKOCN TGUQWTEG KPRWVU CPF C NQY GPXKTQPOGPVCN KORCEV, CU YGNN CU QTICPQNGRVKE, J[IKGPKE CPF GEQPQOKE CFXCPVCIGU HQT RTQFWEGT CPF EQPUWOGT. ÷WTVJGTOQTG, YKPG•U OQUV GPEJCPVKPI CPF HCUEKPCVKPI CURGEV, KVU FKXGTUKV[ QH UV[NG, UJQWNF PGXGT DG VJTGCVGPGF D[ VJG WUG QH VCKNQTGF ITCRGXKPGU CPF YKPG [GCUVU. +P HCEV, IGPG VGEJPQNQI[ UJQWNF TCVJGT DG JCTPGUUGF VQ expand VJG FKXGTUKV[ QH JKIJ SWCNKV[ YKPGU. 6JGUG CTG XGT[ NCTIG EJCNNGPIGU; QXGTEQOKPI VJGO YKNN TGSWKTG C UWDUVCPVKCN CPF UWUVCKPGF GHHQTV. *CRRKN[ VJG TGYCTFU HQT UWEEGUU CTG EQTTGURQPFKPIN[ NCTIG.
#EMPQYNGFIGOGPVU + VJCPM /U /CTKUC *QPG[ CPF /T 6GTT[ 2NCPVKPIC HQT ETKVKECN TGCFKPI QH VJKU OCPWUETKRV. 6JKU %JCRVGT TGRTGUGPVU C EQPFGPUGF CPF KPVGITCVGF XGTUKQP QH TGEGPV, EQORTGJGPUKXG TGXKGYU D[ $CWGT CPF 2TGVQTKWU (ÿ000), &W 6QKV CPF 2TGVQTKWU, (ÿ000), þTQUUOCPP CPF 2TGVQTKWU (1üüü), öCODTGEJVU CPF 2TGVQTKWU (ÿ000), 2TGVQTKWU (1üüü, ÿ000, ÿ001, ÿ003), 2TGVQTKWU GV CN. (1üüü, ÿ003), 2TGVQTKWU CPF $CWGT (ÿ00ÿ), 4CKPKGTK CPF 2TGVQTKWU (ÿ000), 8CP 4GPUDWTI CPF 2TGVQTKWU (ÿ000) CPF 8KXKGT CPF 2TGVQTKWU (ÿ000, ÿ00ÿ). &WG VQ VJG NKOKVGF URCEG CNNQECVGF VQ VJKU EJCRVGT, CNN TGHGTGPEGU CTG PQV NKUVGF JGTG. 6JG TGCFGT VJGTGHQTG KU TGHGTTGF VQ VJG QTKIKPCN UEKGPVKHKE CTVKENGU EKVGF KP VJGUG TGXKGY RCRGTU.
13û
+UCM 5. 2TGVQTKWU
þGHGTGPEGU #P &, ýWIJ %5 (1üü3) 7TGC GZETGVKQP CPF WRVCMG D[ YKPG [GCUVU CU CHHGEVGF D[ XCTKQWU HCEVQTU. #O í ©PQN 8KVKE ûû:3û-û0 #PUCPC[ 8, &GSWKP 5, $NQPFKP $, $CTTG 2 (1üü3) %NQPKPI, UGSWGPEG CPF GZRTGUUKQP QH VJG IGPG GPEQFKPI VJG OCNQNCEVKE GP\[OG HTQO Lactococcus lactis. ÷©$5 öGVV 33ÿ:øû-õ0 #PUCPC[ 8, &GSWKP 5, %COCTCUC %, 5EJCGHHGT 8, þTKXGV í-2, $NQPFKP $, 5CNOQP í-/, $CTTG 2 (1üüù) /CNQNCEVKE HGTOGPVCVKQP D[ GPIKPGGTGF Saccharomyces cerevisiae CU EQORCTGF YKVJ GPIKPGGTGF Schizosaccharomyces pombe. ðGCUV 1ÿ:ÿ1ó-ÿÿó #TOUVTQPI þý, öCODTGEJVU /þ, /CPUXGNV ©2þ, 8CP 8GNFGP &2, 2TGVQTKWU +5 (ÿ001) 9KPG CPF JGCNVJ. 5 #HT í 5EK üø:ÿøü-ÿõÿ #XGT[ 58, *QYNGVV òþ, 4CFKEG 5 (1üüù) %QRRGT VQZKEKV[ VQYCTFU Saccharomyces cerevisiae: FGRGPFGPEG QP RNCUOC OGODTCPG HCVV[ CEKF EQORQUKVKQP. #RRN ©PXKTQP /KETQDKQN ùÿ:3üù0-3üùù $CTTG 2, 8G\KPJGV ÷, &GSWKP 5, $NQPFKP $ (1üü3) þGPGVKE KORTQXGOGPV QH YKPG [GCUV. +P: ÷NGGV þ* (GF) 9KPG /KETQDKQNQI[ CPF $KQVGEJPQNQI[. *CTYQQF #ECFGOKE 2WDNKUJGTU, 5YKV\GTNCPF, RR ûÿ1-ûûø $CWGT ÷÷, 2TGVQTKWU +5 (ÿ000) ðGCUV UVTGUU TGURQPUG CPF HGTOGPVCVKQP GHHKEKGPE[: JQY VQ UWTXKXG VJG OCMKPI QH YKPG. 5 #HT í ©PQN 8KVKE ÿ1:ÿø-ó1 $GEMGT íX9 (ÿ00ÿ) 2NCPV FGHGPEG IGPGU GZRTGUUGF KP VQDCEEQ CPF [GCUV. /5E 6JGUKU, 5VGNNGPDQUEJ 7PKXGTUKV[, 5VGNNGPDQUEJ, 5QWVJ #HTKEC$GIIU í& (1üøõ) 6TCPUHQTOCVKQP QH [GCUV D[ C TGRNKECVKPI J[DTKF RNCUOKF. òCVWTG ÿøó:10û-10ü $KFCTF ÷, $NQPFKP $, &GSWKP 5, 8G\KPJGV ÷, $CTTG 2 (1üüû) %NQPKPI CPF CPCN[UKU QH C FLO5 HNQEEWNCVKQP IGPG HTQO S. cerevisiae. %WTT þGPGV ÿó:1üù-ÿ01 $KUUQP ö÷ (1üüü) 5VWEM CPF UNWIIKUJ HGTOGPVCVKQPU. #O í ©PQN 8KVKE ó0: 10ø-1üüü $KUUQP ö÷, 9CVGTJQWUG #ö, ©DGNGT 5©, 9CNMGT /#, öCRUNG[ í6 (ÿ00ÿ) 6JG RTGUGPV CPF HWVWTG QH VJG KPVGTPCVKQPCN YKPG KPFWUVT[. òCVWTG û1õ:ùüù-ùüü $QNGU ©, *QNNGPDGTI %2 (1üüø) 6JG OQNGEWNCT IGPGVKEU QH JGZQUG VTCPURQTV KP [GCUVU. ÷©/5 /KETQDKQN 4GX ÿ1: õó-111 $QP[ /, $KFCTV ÷, %COCTCUC %, #PUCPC[ 8, &WNCW ö, $CTTG 2, &GSWKP 5 (1üüø) /GVCDQNKE CPCN[UKU QH S. cerevisiae UVTCKPU GPIKPGGTGF HQT OCNQNCEVKE HGTOGPVCVKQP. ÷©$5 öGVV û10:ûóÿ-ûóù $QQPG %, 5FKEW #/, 9CIPGT í, &GITo 4, 5CPEJG\ %, $WUUG[ * (1üü0) +PVGITCVKQP QH VJG [GCUV î1 MKNNGT VQZKP IGPG KPVQ VJG IGPQOG QH OCTMGF YKPG [GCUVU CPF KVU GHHGEV QP XKPKHKECVKQP. #O í ©PQN 8KVKE û1:3ø-ûÿ $QWNVQP $, 5KPINGVQP 8ö, $KUUQP ö÷, îWPMGG 4© (1üüù) ðGCUV CPF DKQEJGOKUVT[ QH GVJCPQN HGTOGPVCVKQP. +P: $QWNVQP $, 5KPINGVQP 8ö, $KUUQP ö÷, îWPMGG 4© (GFU) 2TKPEKRNGU CPF 2TCEVKEGU QH 9KPGOCMKPI. %JCROCP CPF *CNN, òGY ðQTM, RR 13ü-1øÿ %CPCN-öNCWDGTGU 4-/ (1üü3) ©P\[OGU KP YKPGOCMKPI. +P: ÷NGGV þ* (GF) 9KPG /KETQDKQNQI[ CPF $KQVGEJPQNQI[. *CTYQQF #ECFGOKE 2WDNKUJGTU, 5YKV\GTNCPF, RR ûøø-ó0ù %CRWVK # íT, 4[CP 6 (1üüù) /WUV CPF YKPG CEKFKHKECVKQP. 2TGUGPVCVKQP CV C OGGVKPI QH VJG ý+8 ©ZRGTV þTQWR 6GEJPQNQIKG FW 8KP, 2CTKU %CTUVGPU ©, öCODTGEJVU /þ, 2TGVQTKWU +5 (1üüõ) ÷NQEEWNCVKQP, RUGWFQJ[RJCN FGXGNQROGPV CPF KPXCUKXG ITQYVJ KP EQOOGTEKCN YKPG [GCUV UVTCKPU. 5 #HT í ©PQN 8KVKE 1ü:óÿ-ù1 %CTUVGPU /, 8KXKGT /#, 8CP 4GPUDWTI 2, 2TGVQTKWU +5 (ÿ003) ýXGTGZRTGUUKQP, UGETGVKQP CPF CPVKHWPICN CEVKXKV[ QH VJG Saccharomyces cerevisiae EJKVKPCUG. #PP /KETQDKQN (KP RTGUU)
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
13ó
%QNCITCPFG ý, 5KNXC #, ÷WOK /& (1üüû) 4GEGPV CRRNKECVKQPU QH DKQVGEJPQNQI[ KP YKPG RTQFWEVKQP. $KQVGEJPQN 2TQI 10:ÿ-1õ %QNG 8%, òQDNG #% (1üüó) ÷NCXQWT EJGOKUVT[ CPF CUUGUUOGPV. +P: öGC #þ*, 2KIIQVV í4 (GFU) ÷GTOGPVGF $GXGTCIG 2TQFWEVKQP. $NCEMKG #ECFGOKE CPF 2TQHGUUKQPCN, öQPFQP, RR 3ù1-3õó %TQWU í/, 2TGVQTKWU +5, 8CP <[N 9* (1üüù) %NQPKPI CPF GZRTGUUKQP QH VJG ½-öCTCDKPQHWTCPQUKFCUG IGPG (ABF2) QH Aspergillus niger KP Saccharomyces cerevisiae. #RRN /KETQDKQN $KQVGEJPQN ûù:ÿóù-ÿù0 %TQWU í/, 8CP <[N 9*, 2TGVQTKWU +5 (1üüó) %NQPKPI CPF GZRTGUUKQP QH CP Aspergillus kawachii GPFQ-1,û-¾-Z[NCPCUG IGPG KP Saccharomyces cerevisiae. %WTT þGPGV ÿõ:ûùøûø3 &G $CTTQU öQRGU /, 4GJOCP #-7, þQEMQYKCM *, *GKPTKEJ #í, öCPITKFIG 2, *GPUEJMG 2# (ÿ000) ÷GTOGPVCVKQP RTQRGTVKGU QH C YKPG [GCUV QXGTGZRTGUUKPI VJG Saccharomyces cerevisiae IN[EGTQN 3-RJQURJCVG FGJ[FTQIGPCUG IGPG (GPD2). #WUV í þTCRG 9KPG 4GU ù:ÿ0õ-ÿ1ó &GITo 4 (1üü3) 5GNGEVKQP CPF EQOOGTEKCN EWNVKXCVKQP QH YKPG [GCUV CPF DCEVGTKC. +P: ÷NGGV þ* (GF) 9KPG /KETQDKQNQI[ CPF $KQVGEJPQNQI[. *CTYQQF #ECFGOKE 2WDNKUJGTU, 5YKV\GTNCPF, RR ûÿ1-ûûø &GPC[TQNNGU /, #KING /, öQPXCWF-÷WPGN # (1üüó) ÷WPEVKQPCN GZRTGUUKQP KP Saccharomyces cerevisiae QH VJG Lactococcus lactis mle5 IGPG GPEQFKPI VJG OCNQNCEVKE GP\[OG. ÷©/5 /KETQDKQN öGVV 1ÿó:3ø-ûû &GSWKP 5 (ÿ001) 6JG RQVGPVKCN QH IGPGVKE GPIKPGGTKPI HQT KORTQXKPI DTGYKPI, YKPG-OCMKPI CPF DCMKPI [GCUVU. #RRN /KETQDKQN $KQVGEJPQN óù:óøø-óõõ &GSWKP 5, $CTTG 2 (1üüû) /KZGF NCEVKE CEKF-CNEQJQNKE HGTOGPVCVKQP D[ Saccharomyces cerevisiae GZRTGUUKPI VJG Lactobacillus casei ö(ë)-ö&*. $KQñ6GEJPQN 1ÿ:1ø3-1øø &GSWKP 5, $CRVKUVC ©, $CTTG 2 (1üüü) #EKFKHKECVKQP QH ITCRG OWUVU D[ Saccharomyces cerevisiae YKPG [GCUV UVTCKPU IGPGVKECNN[ GPIKPGGTGF VQ RTQFWEG NCEVKE CEKF. #O í ©PQN 8KVKE ó0:ûó-ó0 &QPCNKGU 7©$, 5VCJN 7 (ÿ00ÿ) +PETGCUKPI UWNRJKVG HQTOCVKQP KP Saccharomyces cerevisiae D[ QXGTGZRTGUUKQP QH MET14 CPF SSU1. ðGCUV 1ü:ûøó-ûõû &W 6QKV %, &W 6QKV /, 4CWVGPDCEJ /, 8CP 4GPUDWTI 2, 2TGVQTKWU +5 (ÿ003) 6JG CRRNKECVKQP QH PKUKP, RGFKQEKP CPF NGWEQEKP CU DKQRTGUGTXCVKXGU KP YKPGOCMKPI. +PV í ÷QQF /KETQDKQN (KP RTGUU) &W 6QKV /, 2TGVQTKWU +5 (ÿ000) /KETQDKCN URQKNCIG CPF RTGUGTXCVKQP QH YKPG: WUKPI YGCRQPU HTQO PCVWTG•U QYP CTUGPCN. 5 #HT í ©PQN 8KVKE ÿ1:øû-üù ©INKPVQP í/, *GKPTKEJ #í, 2QNNPKV\ #2, öCPITKFIG 2, *GPUEJMG 2#, &G $CTTQU öQRGU / (ÿ00ÿ) &GETGCUKPI CEGVKE CEKF CEEWOWNCVKQP D[ C IN[EGTQN QXGTRTQFWEKPI UVTCKP QH Saccharomyces cerevisiae D[ FGNGVKPI VJG ALD6 CNFGJ[FG FGJ[FTQIGPCUG IGPG. ðGCUV 1ü:ÿüó-301 ÷QIGN 5, 9GNEJ í9, %CVJCNC þ, îCTKP / (1üõ3) þGPG CORNKHKECVKQP KP [GCUV: CUP1 EQR[ PWODGT TGIWNCVGU EQRRGT TGUKUVCPEG. %WTT þGPGV ø:3ûø-3óó ÷WIGNUCPI î% (1üüø) 9KPG /KETQDKQNQI[. %JCROCP CPF *KNN, òGY ðQTM ÷WINUCPI %%, íQJCPUGP %, %JTKUVICW 5, #FNGT-òKUUGP í (1üüó) #PVKOKETQDKCN GP\[OGU: #RRNKECVKQPU CPF HWVWTG RQVGPVKCN KP VJG HQQF KPFWUVT[. 6TGPFU ÷QQF 5EK 6GEJPQN ù:3ü03üù ÷WLKK 6, îQDC[CUJK ý, ðQUJQOQVQ *, ÷WTWMCYC 5, 6COCK ð (1üüø) ©HHGEV QH CGTCVKQP CPF WPUCVWTCVGF HCVV[ CEKFU QP GZRTGUUKQP QH VJG Saccharomyces cerevisiae CNEQJQN CEGV[NVTCPUHGTCUG IGPG. #RRN ©PXKTQP /KETQDKQN ù3:ü10-ü1ó
13ù
+UCM 5. 2TGVQTKWU
÷WLKK 6, òCICUCYC ò, +YCOCVUW #, $QICMK 6, 6COCK ð, *COCEJK / (1üüû) /QNGEWNCT ENQPKPI, UGSWGPEG CPCN[UKU, CPF GZRTGUUKQP QH VJG [GCUV CNEQJQN CEGV[NVTCPUHGTCUG IGPG. #RRN ©PXKTQP /KETQDKQN ù0:ÿøõù-ÿøüÿ ÷WLKK 6, ðQUJKOQVQ *, 6COCK ð (1üüù) #EGVCVG GUVGT RTQFWEVKQP D[ Saccharomyces cerevisiae NCEMKPI VJG ATF1 IGPG GPEQFKPI VJG CNEQJQN CEGV[NVTCPUHGTCUG. í ÷GTOGPV $KQGPI õ1:ó3õ-óûÿ þCIKCPQ /, $CWGT ÷÷, 2TGVQTKWU +5 (ÿ00ÿ) 6JG UGPUKPI CPF UKIPCNNKPI QH PWVTKVKQPCN UVCVWU CPF VJG TGNCVKQPUJKR VQ HKNCOGPVQWU ITQYVJ KP VJG [GCUV Saccharomyces cerevisiae. ÷©/5 ðGCUV 4GU ÿ:û33-ûø0 þCIKCPQ /, 8CP &[M &, $CWGT ÷÷, öCODTGEJVU /þ, 2TGVQTKWU +5 (1üüüC) /UP1Rñ/UU10R, /UU11R CPF /WE1R CTG RCTV QH C UKIPCN VTCPUFWEVKQP RCVJYC[ FQYPUVTGCO QH /GRÿR TGIWNCVKPI KPXCUKXG ITQYVJ CPF RUGWFQJ[RJCN FKHHGTGPVKCVKQP KP Saccharomyces cerevisiae. /QN /KETQDKQN 31:103-11ù þCIKCPQ /, 8CP &[M &, $CWGT ÷÷, öCODTGEJVU /þ, 2TGVQTKWU +5 (1üüüD) &KXGTIGPV TGIWNCVKQP QH VJG GXQNWVKQPCT[ ENQUGN[ TGNCVGF RTQOQVGTU QH VJG Saccharomyces cerevisiae STA2 CPF MUC1 IGPGU. í $CEVGTKQN 1õ1:ùûüø-ùó0õ þCKPXQTU 5#, ÷To\KGT 8, öGOCTGUSWKGT *, öGSWCTV %, #KING /, $GNCTDK # (1üüû) &GVGEVKQP QH RQN[ICNCEVWTQPCUG, RGEVKP-N[CUG CPF RGEVKP-GUVGTCUG CEVKXKVKGU KP C Saccharomyces cerevisiae UVTCKP. ðGCUV 10:1311-131ü þCNNCPFGT í÷ (1üøø) &GCEKFKHKECVKQP QH GCUVGTP VCDNG YKPGU YKVJ Schizosaccharomyces pombe. #O í ©PQN 8KVKE ÿõ:ùó-ùõ þKWFKEK 2, 4QOCPQ 2,
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
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*GPUEJMG 2#, íKTCPGM 8 (1üü3) ðGCUVU /GVCDQNKUO QH PKVTQIGP EQORQWPFU. +P: ÷NGGV þ* (GF) 9KPG /KETQDKQNQI[ CPF $KQVGEJPQNQI[. *CTYQQF #ECFGOKE 2WDNKUJGTU, 5YKV\GTNCPF, RR øø-1ùû *KPPGP #, *KEMU í$, ÷KPM þ4 (1üøõ) 6TCPUHQTOCVKQP QH [GCUV. 2TQE òCVN #ECF 5EK 75# øó:1üÿü-1ü33 +DTCJKO *4, ðCOCFC /, îQDC[CUJK î, îCVQ # (1üüÿ) $CEVGTKEKFCN CEVKQP QH N[UQ\[OG CICKPUV ITCO-PGICVKXG DCEVGTKC FWG VQ KPUGTVKQP QH C J[FTQRJQDKE RGPVCRGRVKFG KPVQ KVU %-VGTOKPWU. $KQUEK $KQVGEJ $KQEJGO óù:13ù1-13ù3 +PITCJCO íö, þW[OQP í÷ (1üù0) 6JG HQTOCVKQP QH JKIJGT CNKRJCVKE CNEQJQNU D[ OWVCPV UVTCKPU QH Saccharomyces cerevisiae. #TEJ $KQEJGO $KQRJ[U õõ:1óø-1ùù +PITCJCO íö, þW[OQP í÷, %TQYGNN ©# (1üù1) 6JG RCVJYC[ QH HQTOCVKQP QH ò-DWV[N CPF PCO[N CNEQJQNU D[ C OWVCPV UVTCKP QH Saccharomyces cerevisiae. #TEJ $KQEJGO $KQRJ[U üó:1ùü-1øó +UJKFC-÷WLKK î, þQVQ 5, 5WIK[COC *, 6CICMK ð, 5CKMK 6, 6CICMK / (1üüõ) $TGGFKPI QH HNQEEWNGPV KPFWUVTKCN [GCUV UVTCKPU D[ UGNH-ENQPKPI QH VJG HNQEEWNCVKQP IGPG FLO1 CPF TGRGCVGF-DCVEJ HGTOGPVCVKQP D[ VTCPUHQTOCPVU. í þGP #RRN /KETQDKQN ûû:3ûø-3ó3 +XQTTC %, 2GTG\-ýTVKP í©, &GN ýNOQ /+ (1üüü) #P KPXGTUG EQTTGNCVKQP DGVYGGP UVTGUU TGUKUVCPEG CPF UVWEM HGTOGPVCVKQPU KP YKPG [GCUVU. # OQNGEWNCT UVWF[. $KQVGEJPQN $KQGPI ùû:ùüõ-ø0õ îKO í, #NK\CFGJ 2, *CTFKPI 6, *GHPGT-þTCXKPM #, îNKQPUM[ &í (1üüù) &KUTWRVKQP QH VJG [GCUV ATH1 IGPG EQPHGTU DGVVGT UWTXKXCN CHVGT FGJ[FTCVKQP, HTGG\KPI, CPF GVJCPQN UJQEM: RQVGPVKCN EQOOGTEKCN CRRNKECVKQPU. #RRN ©PXKTQP /KETQDKQN ùÿ:1óù3-1óùü îKVCOQVQ î, ýFC î, þQOK î, 6CMCJCUJK î (1üü1) þGPGVKE GPIKPGGTKPI QH C UCMG [GCUV RTQFWEKPI PQ WTGC D[ UWEEGUUKXG FKUTWRVKQP QH CTIKPCUG IGPG. #RRN ©PXKTQP /KETQDKQN óø:301-30ù îTWEMGDGTI #ö (1üüù) 6JG JGZQUG VTCPURQTVGT HCOKN[ QH Saccharomyces cerevisiae. #TEJ /KETQDKQN 1ùù: ÿõ3-ÿüÿ öC þTCPIG &%, %NCG[UUGPU +/, 2TGVQTKWU +5, 8CP <[N 9* (ÿ001) &GITCFCVKQP QH Z[NCP VQ &-Z[NQUG D[ TGEQODKPCPV Saccharomyces cerevisiae EQ-GZRTGUUKPI VJG Aspergillus niger ¾-Z[NQUKFCUG (xlnD) CPF VJG Trichoderma reesei Z[NCPCUG ++ (xyn2) IGPGU. #RRN ©PXKTQP /KETQDKQN ùø:óó1ÿ-óó1ü öC þTCPIG &%, 2TGVQTKWU +5, 8CP <[N 9* (1üüù) ©ZRTGUUKQP QH VJG Trichoderma reesei ¾Z[NCPCUG IGPG (XYN2) KP Saccharomyces cerevisiae. #RRN ©PXKTQP /KETQDKQN ùÿ:103ù10ûû öC þTCPIG &%, 2TGVQTKWU +5, 8CP <[N 9* (1üüø) %NQPKPI QH VJG Bacillus pumilus ¾Z[NQUKFCUG IGPG (xynB) CPF KVU GZRTGUUKQP KP Saccharomyces cerevisiae. #RRN /KETQDKQN $KQVGEJPQN ûø:ÿùÿ-ÿùù öCDCTTG %, þW\\Q í, %CXKP í-÷, &KXKpU % (1üüù) %NQPKPI CPF EJCTCEVGTKUCVKQP QH VJG IGPGU GPEQFKPI VJG OCNQNCEVKE GP\[OG CPF VJG OCNCVG RGTOGCUG QH Leuconostoc oenos. #RRN ©PXKTQP /KETQDKQN ùÿ:1ÿøû-1ÿõÿ öCKPI ©, 2TGVQTKWU +5 (1üüÿ) 5[PVJGUKU CPF UGETGVKQP QH CP Erwinia chrysanthemi RGEVCVG N[CUG KP Saccharomyces cerevisiae TGIWNCVGF D[ FKHHGTGPV EQODKPCVKQPU QH DCEVGTKCN CPF [GCUV RTQOQVGT CPF UKIPCN UGSWGPEGU. þGPG 1ÿ1:3ó-ûó öCKPI ©, 2TGVQTKWU +5 (1üü3C) 6JG RTKOCT[ UVTWEVWTG CPF GZRTGUUKQP QH CP Erwinia carotovora RQN[ICNCEVWTQPCUG-GPEQFKPI IGPG (peh1) KP Escherichia coli CPF [GCUV. í #RRN $CEVGTKQN øó:1ûü-1óõ
13õ
+UCM 5. 2TGVQTKWU
öCKPI ©, 2TGVQTKWU +5 (1üü3D) %Q-GZRTGUUKQP QH CP Erwinia chrysanthemi RGEVCVG N[CUGGPEQFKPI IGPG (pelE) CPF CP Erwinia carotovora RQN[ICNCEVWTQPCUG-GPEQFKPI IGPG (peh1) KP Saccharomyces cerevisiae. #RRN /KETQDKQN $KQVGEJPQN 3ü:1õ1-1õõ öCODTGEJVU /þ, 2TGVQTKWU +5 (ÿ000) ðGCUV CPF KVU KORQTVCPEG VQ YKPG CTQOC. 5 #HT í ©PQN 8KVKE ÿ1:üø-1ÿü öCODTGEJVU /þ, $CWGT ÷÷, /CTOWT í, 2TGVQTKWU +5 (1üüù) /WE1, C OWEKP-NKMG RTQVGKP VJCV KU TGIWNCVGF D[ MSS10, KU ETKVKECN HQT RUGWFQJ[RJCN FKHHGTGPVKCVKQP KP [GCUV. 2TQE òCVN #ECF 5EK 75# ü3:õû1ü-õûÿû öKNN[ /, öCODTGEJVU /þ, 2TGVQTKWU +5 (ÿ000) 6JG GHHGEV QH KPETGCUGF [GCUV CNEQJQN CEGV[NVTCPUHGTCUG CEVKXKV[ QP VJG UGPUQTKCN SWCNKV[ QH YKPG CPF DTCPF[. #RRN ©PXKTQP /KETQDKQN ùù:øûû-øó3 öQ 9-5, &TCPIKPKU #/ (1üüù) FLO11, C [GCUV IGPG TGNCVGF VQ VJG STA IGPGU, GPEQFGU C PQXGN EGNN UWTHCEG HNQEEWNKP. í $CEVGTKQN 1øõ:ø1ûû-ø1ó1 öWVVKI /, 2TGVQTKWU +5, 8CP <[N 9* (1üüø) %NQPKPI QH VYQ ¾-Z[NCPCUG-GPEQFKPI IGPGU HTQO Aspergillus niger CPF VJGKT GZRTGUUKQP KP Saccharomyces cerevisiae. $KQVGEJPQN öGVV 1ü:û11-û1ó öW[VGP, î, 4KQW, %, $NQPFKP, $ (ÿ00ÿ) 6JG JGZQUG VTCPURQTVGTU QH Saccharomyces cerevisiae RNC[ FKHHGTGPV TQNGU FWTKPI GPQNQIKECN HGTOGPVCVKQP. ðGCUV 1ü: ø13-øÿù ö[PF ö4, 9GKOGT 2í, 8CP <[N 9*, 2TGVQTKWU +5 (ÿ00ÿ) /KETQDKCN EGNNWNQUG WVKNK\CVKQP: HWPFCOGPVCNU CPF DKQVGEJPQNQI[. /KETQDKQN /QN $KQN 4GX ùù:ó0ù-óøø /CNEQTRU 2, &WHQWT í2 (1üõø) ©UVGT U[PVJGUKU D[ Saccharomyces cerevisiae: öQECNKUCVKQP QH CEGV[N-%Q#: +UQ-CO[NCNEQJQN CEGV[NVTCPUHGTCUG (É#6Ì). 2TQE ©WT $TGY %QPX ÿ1:3øø-3õû /CNEQTRU 2, %JGXCN í/, íCOKN 5, &WHQWT í2 (1üü1) # PGY OQFGN HQT VJG TGIWNCVKQP QH GUVGT U[PVJGUKU D[ CNEQJQN CEGV[NVTCPUHGTCUG KP Saccharomyces cerevisiae FWTKPI HGTOGPVCVKQP. í #O 5QE $TGY %JGO ûü:ûø-ó3 /CNJGTDG &÷, &W 6QKV /, %QTFGTQ ýVGTQ 44, 8CP 4GPUDWTI 2, 2TGVQTKWU +5 (ÿ00ÿ) ©ZRTGUUKQP QH VJG Aspergillus niger INWEQUG QZKFCUG IGPG (GOX1) KP Saccharomyces cerevisiae CPF KVU RQVGPVKCN CRRNKECVKQP KP YKPG RTQFWEVKQP. #RRN /KETQDKQN $KQVGEJPQN (KP RTGUU) /KEJPKEM 5, 4QWUVCP í-ö, 4GOK\G ÷, $CTTG 2, &GSWKP 5 (1üüø) /QFWNCVKQP QH IN[EGTQN CPF GVJCPQN [KGNFU FWTKPI CNEQJQNKE HGTOGPVCVKQP KP Saccharomyces cerevisiae UVTCKPU QXGTGZRTGUUGF QT FKUTWRVGF HQT GPD1 GPEQFKPI IN[EGTQN 3-RJQURJCVG FGJ[FTQIGPCUG. ðGCUV 13:øõ3-øü3 òCMCOWTC 5, 6CMCUCMK *, îQDC[CUJK î, îCVQ # (1üü3) *[RGTIN[EQU[NCVKQP QH JGP GII YJKVG N[UQ\[OG KP [GCUV. í $KQN %JGO ÿùõ:1ÿø0ù-1ÿø1ÿ òQDNG #% (1üüû) 9KPG HNCXQWT. +P: 2KIIQVV í4, 2CVVGTUQP # (GFU) 7PFGTUVCPFKPI òCVWTCN ÷NCXQWTU. $NCEMKG CPF 2TQHGUUKQPCN, þNCUIQY, RR ÿÿõ-ÿûÿ ýNKXGT 5þ (1üüù) ÷TQO &ò# UGSWGPEG VQ DKQNQIKECN HWPEVKQP. òCVWTG 3øü:óüø-ù00 ·UVGTICCTF 5, ýNUUQP ö, òKGNUGP í (ÿ000) /GVCDQNKE GPIKPGGTKPI QH Saccharomyces cerevisiae. /KETQDKQN /QN $KQN 4GX ùû: 3û-ó0 ýWIJ %5, %TQYGN ©#, þWVNQXG $4 (1üõõ) %CTDCO[N EQORQWPF TGCEVKQPU YKVJ GVJCPQN. #O í ©PQN 8KVKE 3ü:ÿ3ü-ÿûÿ f\ECP 5, íQJPUVQP / (1üüü) ÷WPEVKQP CPF TGIWNCVKQP QH [GCUV JGZQUG VTCPURQTVGTU. /KETQDKQN /QN $KQN 4GX ù3: óóû-óùü 2oTG\-þQP\hNG\ í#, &G þTCCH ö*, 8KUUGT í, 4COQP & (1üüù) /QNGEWNCT ENQPKPI CPF GZRTGUUKQP KP Saccharomyces cerevisiae QH VYQ Aspergillus nidulans Z[NCPCUG IGPGU. #RRN ©PXKTQP /KETQDKQN ùÿ:ÿ1øü-ÿ1õÿ
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
13ü
2oTG\-þQP\hNG\ í#, þQP\hNG\ 4, úWGTQN #, 5GPFTC í, 4COQP & (1üü3) %QPUVTWEVKQP QH C TGEQODKPCPV YKPG [GCUV UVTCKP GZRTGUUKPI ¾-(1,û)-GPFQINWECPCUG CPF KVU WUG KP OKETQXKPKHKECVKQP RTQEGUUGU. #RRN ©PXKTQP /KETQDKQN óü:ÿõ01-ÿõ0ù 2GVGTUQP 5*, 8CP <[N 9*, 2TGVQTKWU +5 (1üüõ) &GXGNQROGPV QH C RQN[UCEEJCTKFG-FGITCFKPI UVTCKP QH Saccharomyces cerevisiae. $KQVGEJPQN 6GEJ 1ÿ:ù1ó-ù1ü 2KEMGTKPI þ (1üüõ) 6JG WUG QH GP\[OGU VQ UVCDKNKUG EQNQWT CPF HNCXQWT KP YKPG - CP CNVGTPCVKXG VQ 5ýÿ. #WUV þTCRGITQYGT ê 9KPGOCMGT 5GRVGODGT:101-103 2KEMGTKPI þ, *GCVJGTDGNN &# (1üüù) %JCTCEVGTKUCVKQP QH TGFWEGF CNEQJQN YKPG OCFG HTQO INWEQUG QZKFCUG VTGCVGF OWUV. ÷QQF 6GEJPQN ÿù:101-10ø 2KEMGTKPI þí, *GCVJGTDGNN &#, $CTPGU /÷ (1üüõ) ýRVKOKUKPI INWEQUG EQPXGTUKQP KP VJG RTQFWEVKQP QH TGFWEGF CNEQJQN YKPG WUKPI INWEQUG QZKFCUG. ÷QQF 4GU +PV 31:ùõó-ùüÿ 2KEMGTKPI þí, *GCVJGTDGNN &#, $CTPGU /÷ (1üüüC) 6JG RTQFWEVKQP QH TGFWEGF-CNEQJQN YKPG WUKPI INWEQUG QZKFCUG VTGCVGF LWKEG. 2CTV + %QORQUKVKQP. #O í ©PQN 8KVKE ó0:ÿü1-ÿüõ 2KEMGTKPI þí, *GCVJGTDGNN &#, $CTPGU /÷ (1üüüD) 6JG RTQFWEVKQP QH TGFWEGF-CNEQJQN YKPG WUKPI INWEQUG QZKFCUG VTGCVGF LWKEG. 2CTV ++ 5VCDKNKV[ CPF 5ýÿ-DKPFKPI. #O í ©PQN 8KVKE ó0:ÿüü-30ù 2KEMGTKPI þí, *GCVJGTDGNN &#, $CTPGU /÷ (1üüüE). 6JG RTQFWEVKQP QH TGFWEGF-CNEQJQN YKPG WUKPI INWEQUG QZKFCUG VTGCVGF LWKEG. 2CTV +++ 5GPUQT[. #O í ©PQN 8KVKE ó0:30ø-31ù 2QTTQ &ö, $TCODKNNC ö, 4CP\K $/, /CTVGICPK ©, #NDGTIJKPC ö (1üüó) &GXGNQROGPV QH OGVCDQNKECNN[ GPIKPGGTGF Saccharomyces cerevisiae EGNNU HQT VJG RTQFWEVKQP QH NCEVKE CEKF. $KQVGEJPQN 2TQI 11:ÿüû-ÿüõ 2TGVQTKWU +5 (1üüø) 7VKNK\CVKQP QH RQN[UCEEJCTKFGU D[ Saccharomyces cerevisiae. +P:
1û0
+UCM 5. 2TGVQTKWU
4CWJWV & (1üü3) ðGCUV 2TQFWEVKQP QH UWNRJWT EQORQWPFU. +P: ÷NGGV þ* (GF) 9KPG /KETQDKQNQI[ CPF $KQVGEJPQNQI[. *CTYQQF #ECFGOKE 2WDNKUJGTU, 5YKV\GTNCPF, RR 1õ3ÿÿ3 4GOK\G ÷, 4QWUVCP íö, 5CDNC[TQNNGU í/, $CTTG 2, &GSWKP 5 (1üüü) þN[EGTQN QXGTRTQFWEVKQP D[ GPIKPGGTGF Saccharomyces cerevisiae YKPG [GCUV UVTCKPU NGCFU VQ UWDUVCPVKCN EJCPIGU KP D[-RTQFWEV HQTOCVKQP CPF VQ C UVKOWNCVKQP QH HGTOGPVCVKQP TCVG KP UVCVKQPCT[ RJCUG. #RRN ©PXKTQP /KETQDKQN ùó:1û3-1ûü 4KDoTGCW-þC[QP 2, &WDQWTFKGW &, &QPoEJG $, öQPXCWF # (ÿ000) *CPFDQQM QH ©PQNQI[. 6JG /KETQDKQNQI[ QH 9KPG CPF 8KPKHKECVKQPU, 8QN 1. íQJP 9KNG[ ê 5QPU öVF, %JKEJGUVGT 5CNOQP í-/, $CTTG 2 (1üüõ) +ORTQXGOGPV QH PKVTQIGP CUUKOKNCVKQP CPF HGTOGPVCVKQP MKPGVKEU WPFGT GPQNQIKECN EQPFKVKQPU D[ FGTGRTGUUKQP QH CNVGTPCVKXG PKVTQIGP-CUUKOKNCVQT[ RCVJYC[U KP CP KPFWUVTKCN Saccharomyces cerevisiae UVTCKP. #RRN ©PXKTQP /KETQDKQN ùû:3õ31-3õ3ø 5ECPGU î6, *QJOCPP 5, 2TKQT $# (1üüõ) þN[EGTQN RTQFWEVKQP D[ VJG [GCUV Saccharomyces cerevisiae CPF KVU TGNGXCPEG VQ YKPG: # TGXKGY. 5 #HT í ©PQN 8KVKE 1ü:1ø-ÿÿ 5EJOKVV /í, îNCXGJP 2, 9CPI í, 5EJ{PKI +, 6KRRGT &í (1üüù) %GNN E[ENG UVWFKGU QP VJG OQFG QH CEVKQP QH [GCUV îÿõ MKNNGT VQZKP. /KETQDKQNQI[ (7î) 1ûÿ:ÿùóó-ÿùùÿ 5EJQGOCP *, 8KXKGT /#, &W 6QKV /, &KEMU ö/6, 2TGVQTKWU +5 (1üüü) 6JG FGXGNQROGPV QH DCEVGTKEKFCN [GCUV UVTCKPU D[ GZRTGUUKPI VJG Pediococcus acidilactici RGFKQEKP IGPG (pedA) KP Saccharomyces cerevisiae. ðGCUV 1ó:ùûø-ùóù 5EWFCOQTG-5OKVJ 2, /QTCP í (1üüø) # ITQYKPI OCTMGV HQT TGFWEGF CNEQJQN YKPGU. 9KPG +PF í 1ÿ:1ùó-1ùø 5JKOC í, *KPQ #, ðCOCFC-+[Q %, 5W\WMK ð, òCMCLKOC 4, 9CVCPCDG *, /QTK î, 6CMCPQ * (1üüü) 5VTGUU VQNGTCPEG KP FQWIJU QH Saccharomyces cerevisiae VTGJCNCUG OWVCPVU FGTKXGF HTQO EQOOGTEKCN DCMGT•U [GCUV. #RRN ©PXKTQP /KETQDKQN ùó:ÿõû1-ÿõûù 5JKOK\W î (1üü3) îKNNGT [GCUVU. +P: ÷NGGV þ* (GF) 9KPG /KETQDKQNQI[ CPF $KQVGEJPQNQI[. *CTYQQF #ECFGOKE 2WDNKUJGTU, 5YKV\GTNCPF, RR ÿû3-ÿùû 5OKV #, %QTFGTQ ýVGTQ 44, öCODTGEJVU /þ, 2TGVQTKWU +5, 8CP 4GPUDWTI 2 (ÿ003) /CPKRWNCVKQP QH XQNCVKNG RJGPQN EQPEGPVTCVKQPU KP YKPG D[ GZRTGUUKPI XCTKQWU RJGPQNKE CEKF FGECTDQZ[NCUG IGPGU KP Saccharomyces cerevisiae. #RRN /KETQDKQN $KQVGEJPQN (KP RTGUU) 5OKVU *2, *CWH í, /ôNNGT 5, *QDNG[ 6í,
û 6JG IGPGVKE CPCN[UKU CPF VCKNQTKPI QH YKPG [GCUVU
1û1
6GWPKUUGP #, &WOQTVKGT ÷, þQTYC /-÷, $CWGT í, 6CPIJG #, öQvG\ #, 5OGV 2, 8CP &KLEM 2, 6JGXGNGKP í/ (ÿ00ÿ) +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH C HTGG\G-VQNGTCPV FKRNQKF FGTKXCVKXG QH CP KPFWUVTKCN DCMGT•U [GCUV UVTCKP CPF KVU WUG KP HTQ\GP FQWIJU. #RRN ©PXKTQP /KETQDKQN ùõ:ûøõ0-ûøõø 6GWPKUUGP #9, 5VGGPUOC *ð (1üüó) 4GXKGY: 6JG FQOKPCPV HNQEEWNCVKQP IGPGU QH Saccharomyces cerevisiae EQPUVKVWVG C PGY UWDVGNQOGTKE IGPG HCOKN[. ðGCUV 11:1001-1013 6JGXGNGKP í/ (1üüù) 4GIWNCVKQP QH VTGJCNQUG OGVCDQNKUO CPF KVU TGNGXCPEG VQ EGNN ITQYVJ CPF HWPEVKQP. +P: $TCODN 4, /CT\NWH þ# (GFU) 6JG /[EQVC +++. 5RTKPIGT-8GTNCI, $GTNKP CPF *GKFGNDGTI, RR 3üó-ûÿ0 6JGXGNGKP í/, &G 9KPFG í* (1üüü) òQXGN UGPUKPI OGEJCPKUOU CPF VCTIGVU HQT VJG E#/2RTQVGKP MKPCUG # RCVJYC[ KP VJG [GCUV Saccharomyces cerevisiae. /QN /KETQDKQN 33:ü0û-ü1õ 6QTTGC þQwK &, #PEsP #\RKNKEWGVC % (ÿ001) +PHNWGPEG QH [GCUV UVTCKP QP DKQIGPKE COKPGU EQPVGPV KP YKPGU: TGNCVKQPUJKR YKVJ VJG WVKNK\CVKQP QH COKPQ CEKFU FWTKPI HGTOGPVCVKQP. #O í ©PQN 8KVKE óÿ:1õó-1ü0 6TQOR #, &G îNGTM %# (1üõõ) ©HHGEV QH EQRRGTQZ[EJNQTKFG QP VJG HGTOGPVCVKQP QH OWUV CPF QP YKPG SWCNKV[. 5 #HT í ©PQN 8KVKE ü:31-3ù 8CFCU\ #5, ÷TCPMGP &$, þQXGPFGT, $ö, íCIICPCVJ &$, þQXGPFGT 2, #TKCVVK /, 2TGVQTKWU +5, þWRVJCT #5 (ÿ00ÿ) 2TQRGTVKGU QH C YKPG [GCUV CPVCIQPKUV, Saccharomyces cerevisiae 6ÿ0ù. 5 #HT í ©PQN 8KVKE ÿ3:3ü-ûø 8CP &KLEM 2, þQTYC /-÷, öGOCKTG î, 6GWPKUUGP #, 8GTUGNG /, %QNQODQ 5, &WOQTVKGT ÷, /C 2, 6CPIJG #, öQvG\ #, 6JGXGNGKP í/ (ÿ000) %JCTCEVGTKUCVKQP QH C PGY UGV QH OWVCPVU FGHKEKGPV KP HGTOGPVCVKQP KPFWEGF NQUU QH UVTGUU TGUKUVCPEG HQT WUG KP HTQ\GP FQWIJ CRRNKECVKQPU. +PV í ÷QQF /KETQDKQN óó:1õø-1üÿ 8CP 4GPUDWTI 2, 2TGVQTKWU +5 (ÿ000) ©P\[OGU KP YKPGOCMKPI: JCTPGUUKPI PCVWTCN ECVCN[UVU HQT GHHKEKGPV DKQVTCPUHQTOCVKQPU. 5 #HT í ©PQN 8KVKE ÿ1:óÿ-ø3 8CP 4GPUDWTI 2, 8CP <[N 9*, 2TGVQTKWU +5 (1üüû) ©ZRTGUUKQP QH VJG Butyrivibrio fibrisolvens GPFQ-¾-1,û-INWECPCUG IGPG VQIGVJGT YKVJ VJG Erwinia RGEVCVG N[CUG CPF RQN[ICNCEVWTQPCUG IGPG KP Saccharomyces cerevisiae. %WTT þGPGV ÿø:1ø-ÿÿ 8CP 4GPUDWTI 2, 8CP <[N 9*, 2TGVQTKWU +5 (1üüó) ©ZRTGUUKQP QH VJG Ruminococcus flavefaciens EGNNQFGZVTKPCUG IGPG KP Saccharomyces cerevisiae. $KQVGEJ öGVV 1ø:ûõ1-ûõù 8CP 4GPUDWTI 2, 8CP <[N 9*, 2TGVQTKWU +5 (1üüù) %Q-GZRTGUUKQP QH C Phanerochaete chrysosporium EGNNQDKQJ[FTQNCUG IGPG CPF C Butyrivibrio fibrisolvens GPFQ-¾-1,ûINWECPCUG IGPG KP Saccharomyces cerevisiae. %WTT þGPGV 30:ÿûù-ÿó0 8CP 4GPUDWTI 2, 8CP <[N 9*, 2TGVQTKWU +5 (1üüø) ýXGT-GZRTGUUKQP QH VJG Saccharomyces cerevisiae GZQ-¾-1,3-INWECPCUG IGPG VQIGVJGT YKVJ VJG Bacillus subtilis GPFQ-¾-1,3-1,ûINWECPCUG IGPG CPF VJG Butyrivibrio fibrisolvens GPFQ-¾-1,û-INWECPCUG IGPG KP [GCUV. í $KQVGEJPQN óó:û3-ó3 8CP 4GPUDWTI 2, 8CP <[N 9*, 2TGVQTKWU +5 (1üüõ) ©PIKPGGTKPI [GCUV HQT GHHKEKGPV EGNNWNQUG FGITCFCVKQP. ðGCUV 1û:ùø-øù 8GTUVTGRGP îí, $CWGT ÷÷, 9KPFGTKEMZ í, >FGNKPEMZ þ, 2TGVQTKWU +5, 6JGXGNGKP í/, &GNXCWZ ÷4 (ÿ001) öCVG HGTOGPVCVKQP GZRTGUUKQP QH FLO1 KP Saccharomyces cerevisiae. í #OGT 5QE $TGY %JGO óÿ:ùü-øù 8KUUGT íí (1üüü) %NQPKPI CPF GZRTGUUKQP QH VJG Lactobacillus fermentum CEKF WTGCUGU IGPG KP Saccharomyces cerevisiae. /5E VJGUKU, 5VGNNGPDQUEJ 7PKXGTUKV[, 5VGNNGPDQUEJ, 5QWVJ #HTKEC 8KXKGT /#, 2TGVQTKWU +5 (ÿ000) þGPGVKE KORTQXGOGPV QH ITCRGXKPG: VCKNQTKPI ITCRG XCTKGVKGU HQT VJG VJKTF OKNNGPPKWO. 5 #HT í ©PQN 8KVKE ÿ1:ó-ÿù
1ûÿ
+UCM 5. 2TGVQTKWU
8KXKGT /#, 2TGVQTKWU +5 (ÿ00ÿ) þGPGVKECNN[ VCKNQTGF ITCRGXKPGU HQT VJG YKPG KPFWUVT[. 6TGPFU $KQVGEJPQN ÿ0:ûøÿ-ûøõ 8QNUEJGPM *, 8KNLQGP /, þTQDNGT í, 2GV\QNF $, $CWGT ÷÷, 5WDFGP 4, ðQWPI 4#, öQPXCWF #, &GPC[TQNNGU /, 8CP 8WWTGP *íí (1üüø) ©PIKPGGTKPI RCVJYC[U HQT OCNCVG FGITCFCVKQP KP Saccharomyces cerevisiae. òCVWTG $KQVGEJ 1ó:ÿó3-ÿóø 9CNMGT þ/ (1üüõ) ðGCUV 2J[UKQNQI[ CPF $KQVGEJPQNQI[. íQJP 9KNG[ CPF 5QPU, òGY ðQTM 9CVCTK í, òQOWTC /, 5CJCTC *, îQUJKPQ 5 (1üü3) %QPUVTWEVKQP QH HNQEEWNGPV DTGYGT•U [GCUV D[ EJTQOQUQOCN KPVGITCVKQP QH VJG [GCUV HNQEEWNCVKQP IGPG FLO1. í +PUV $TGY 100:ø3-øø 9GDDGT #ö, öCODTGEJVU /þ, 2TGVQTKWU +5 (1üüø) MSS11, C PQXGN [GCUV IGPG KPXQNXGF KP VJG TGIWNCVKQP QH UVCTEJ OGVCDQNKUO. %WTT þGPGV 3ÿ:ÿù0-ÿùù 9KNNKCOU 5#, *QFIGU 4#, 5VTKMG 6ö, 5PQY 4, îWPMGG 4© (1üõû) %NQPKPI VJG IGPG HQT VJG OCNQNCEVKE HGTOGPVCVKQP QH YKPG HTQO Lactobacillus delbrueckii KP Escherichia coli CPF [GCUVU. #RRN ©PXKTQP /KETQDKQN ûø:ÿõõ-ÿü3
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV ,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
#DUVTCEV 6JG CTV QH DGGT DTGYKPI KU CPEKGPVþ CPF Saccharomyces [GCUV RTQDCDN[ RNC[GF C RKXQVCN TQNG HTQO VJG DGIKPPKPI. 2TQFWEVKQP QH DGGT HTQO VJG DCTNG[ ITCKP EQPUKUVU QH OWNVKRNG UVGRUþ QH YJKEJ QPN[ VJG NCUV HGY KPXQNXG VJG [GCUV. 0GXGTVJGNGUUþ VJG DGJCXKQWT QH VJG [GCUV KU JKIJN[ FGEKUKXG HQT DQVJ URGGF CPF QWVEQOG QH VJG YJQNG RTQEGUUþ CPF VQ C NCTIG GZVGPV KPHNWGPEGU VJG VQVCN DGGT HNCXQWT RTQHKNG. LQPI MPQYP HGTOGPVCVKQPÿTGNCVGF RTQDNGOU CU YGNN CU VJG QEECUKQPCN GXQNWVKQP QH EGTVCKP QHHÿ HNCXQWTU ECP DG DNCOGF QP VJG [GCUV. )TGCV GHHQTVU JCXG VJGTGHQTG IQPG KPVQ UVWFKGU QH VJG IGPGVKEU QH DTGYKPI [GCUVþ UVCTVKPI YKVJ VJG RWTG EWNVKXCVKQP QH NCIGT DTGYKPI [GCUV OQTG VJCP 1ýý [GCTU CIQ CPF EQPVKPWKPI YKVJ VQFC[•U FGVCKNGF FGUETKRVKQP QH VJG DTGYKPI [GCUV IGPQOGþ CU YGNN CU XGPVWTGU KPVQ IGPGVKE OQFKHKECVKQP VQ UQNXG DTGYKPI RTQDNGOU. *GTG YG FGUETKDG EWTTGPV MPQYNGFIG QP DTGYKPI [GCUV IGPGVKEU CPF RJ[NQIGP[þ HKGNFUþ YJKEJ JCXG FGXGNQRGF YKVJ CP KPETGCUKPI URGGF FWTKPI VJG NCUV EQWRNG QH FGECFGU. (WTVJGTþ YG RQTVTC[ VJG IGPGVKE VGEJPQNQIKGU VJCV CTG CXCKNCDNG HQT VJG [GCUV DTGGFGT KP JKU QT JGT GPFGCXQWTU HQT UVTCKP KORTQXGOGPV. /CjQT CTGCU HQT VJG KORTQXGOGPV QH VJG DTGYKPI [GCUV DGJCXKQWT CTG FKUEWUUGFþ CPF VJG UEKGPVKHKE DCUGU HQT RCUV CPF EWTTGPV GHHQTVU VQ UQNXG VJGUG RTQDNGOU D[ IGPGVKE CRRTQCEJGU CTG FGUETKDGFþ CU YGNN CU VJG UVTCVGIKGU GORNQ[GF. (KPCNN[þ YG URGEWNCVG QP VJG HWVWTG QH DTGYKPI [GCUV OQFKHKECVKQPþ CPF VCMG KPVQ EQPUKFGTCVKQP VJG ITGCV NCPFOCTMU KP [GCUV TGUGCTEJ FWTKPI VJG NCUV EQWRNG QH FGEÿ CFGU. 9G CNUQ DTKGHN[ FKUEWUU OCVVGTU QH RWDNKE RGTEGRVKQP QH IGPGVKECNN[ OQFKHKGF DTGYKPI [GCUV.
5.1 6JG TQNG QH [GCUV KP DGGT RTQFWEVKQP $GGT DTGYKPI KU CP CPEKGPV CTVþ MPQYP CU GCTN[ CU 3ýýý $% HTQO /GUQRQVCOKC üUGGþ G.I.þ %QTTCP 1û75 HQT C TGXKGY QH DGGT DTGYKPI JKUVQT[ú. ùPKÿEGNNWNCT HWPIKþ UQÿECNNGF [GCUVþ YGTG RTQDCDN[ VJG HGTOGPVKPI CIGPV HTQO VJG DGIKPPKPI CU KU UWIÿ IGUVGF HTQO OQTG VJCP 3ýýý [GCTÿQNF HKPFKPIU HTQO É6JG 0GY -KPIFQOÌ KP ©I[RV üSCOWGN 1ûû÷ú. +P EQORCTKUQPþ DGGT DTGYKPI KP ©WTQRG KU C TCVJGT PGY VGEJPQNQIKÿ ECN NCPFOCTM VJCV CRRGCTU VQ DG CEqWKTGF HTQO VJG /KFFNG ©CUV KPFGRGPFGPVN[ D[ )GTOCPKE CPF %GNVKE VTKDGU CTQWPF VJG 1UV EGPVWT[ #& ü%QTTCP 1û75ú. (WTVJGT FGÿ XGNQROGPV QH DGGT DTGYKPI OQUVN[ VQQM RNCEG KP ©PINCPFö$TKVCKP CPF KP VJG Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
144
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
(KI. 1. SEJGOCVKE TGRTGUGPVCVKQP QH VJG HKXG OCKP UVCIGU KP DGGT DTGYKPI.
©WTQRGCP JGCTVNCPFþ KP RCTVKEWNCT $QJGOKC CPF )GTOCP[. 9JKNG QP VJG $TKVKUJ +UNGUþ VJG VTCFKVKQP QH CNG DTGYKPI YCU VCMGP VQ RGTHGEVKQPþ #NG DTGYKPI YCU FGXGNÿ QRGF KPVQ NCIGT DTGYKPI KP OCKPNCPF ©WTQRG. õPG QH VJG OQUV KORQTVCPV VGEJPQÿ NQIKECN OKNGUVQPGU KP DGGT DTGYKPI YCU VJG RWTG EWNVWTKPI QH DTGYKPI [GCUVþ KP HCEV EQPUVKVWVKPI C UJKHV QH RCTCFKIOU HQT VJG GCTN[ ôýVJ EGPVWT[ DTGYGTU. ùR WPVKN VJKU VKOGþ YQTV YCU KPXCTKCDN[ KPQEWNCVGF YKVJ URGPV [GCUV QH VJG RTGEGFKPI HGTOGPVCÿ VKQPþ DWV KP 1883þ ©OKN %JT. *CPUGP QH VJG %CTNUDGTI LCDQTCVQT[ FGXKUGF C OGVJQF HQT WUKPI UKPINGÿEGNN EWNVWTGU QH [GCUV KP DGGT RTQFWEVKQP ü*CPUGP 1883ú. 6JG WUG QH RWTG [GCUV EWNVWTGU YCU qWKEMN[ CFQRVGF YQTNFYKFGþ HKTUV KP NCIGT CPF UWDUGÿ qWGPVN[ KP CNG DTGYKPI. 9JKNG OKZGF EWNVWTGU QH UGXGTCN [GCUV UVTCKPU CTG qWKVG YKFGN[ WUGF KP CNG DTGYKPIþ URQPVCPGQWU HGTOGPVCVKQP QP CP KPFWUVTKCN UECNG KU QPN[ WUGF KP C HGY ECUGUþ G.I. $GNIKCP lambic DTGYKPI.6JG IGPGTCNKUGF UEJGOG QH DGGT DTGYKPI FGUETKDGU VJG HGTOGPVCVKQP QH C ENCTKHKGF FKIGUV QH IGTOKPCVGF DCTNG[ ITCKPU D[ Saccharomyces [GCUVUþ VQ RTQFWEG VJG DGGTþ PQVCDN[ EQPVCKPKPI GVJCPQNþ ECTDQP FKQZKFGþ CPF CTQOC EQORQWPFU. 6JKU UEJGOG OC[ DG FKXKFGF KPVQ HKXG OCKP UVCIGU üUGG (KI. 1ú üUGG *QWIJ GV CN. 1û8ô HQT C VJQTQWIJ FGUETKRVKQP QH DGGT DTGYKPIú. ü1ú $CTNG[ ITCKPU QH C URGEKCN OCNVKPI XCTKGV[ CTG IGTOKPCVGF KP VJG UQÿ ECNNGF ÉOCNVKPIÌ RTQEGUU: 6JG DCTNG[ KU UVGGRGF YKVJ YCVGTþ CNNQYKPI URTQWVKPI QH VJG ITCKPU. 6JKU TGUWNVU KP VJG DKQU[PVJGUKU CPF CEVKXCVKQP QH GPFQIGPQWU GP\[OGUþ YJKEJ CTG VTCPURQTVGF VQ VJG ITCKP GPFQURGTO. $TGCMFQYP QH EGNN YCNN OCVGTKCN GPÿ UWGU. üôú 6JG TGUWNVKPI ÉOCNVÌ KU FTKGFþ OKNNGFþ CPF UWURGPFGF KP YCVGT. ü3ú 6JKU UWURGPUKQP KU JGCVGF KP UVGRYKUG KPETGOGPVU KP VGORGTCVWTG VQ CEVKXCVG ECTDQJ[ÿ FTCVGÿFGITCFKPI CPF RTQVGKPÿFGITCFKPI DCTNG[ GP\[OGU. 6JG OCNV GZVTCEV KU HKNÿ VGTGFþ CPF VJG TGUWNVKPI ÉUYGGV YQTVÌ KU DQKNGF YKVJ JQRU QT JQR GZVTCEV VQ RTQFWEG VJG HKPCN ÉDTGYGT•U YQTVÌ. 6JG YQTV PQTOCNN[ EQPVCKPU OQTG VJCP 1ýó üYöXú QH NQYÿOQNGEWNCTÿYGKIJV UWICTUþ OQUVN[ OCNVQUGþ CPF HQTOU CP KFGCN ITQYVJ OGFKWO HQT VJG RTQRCICVGF DTGYKPI [GCUV CFFGF CV VJKU UVCIG. ü4ú 6JG UQÿECNNGF ÉRTKOCT[ HGTOGPVCVKQPÌ RTQEGGFU HQT C PWODGT QH FC[Uþ FGRGPFKPI QP VJG V[RG QH DGGT RTQÿ
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
145
FWEGF. ü5ú (KPCNN[þ ÉOCVWTCVKQPÌ QT ÉUGEQPFCT[ HGTOGPVCVKQPÌ KU WUWCNN[ PGGFGF VQ CNNQY TGOCKPKPI [GCUV EGNNU VQ RTQFWEG FGUKTGF HNCXQWTU CPFöQT VQ IGV TKF QH QHHÿ HNCXQWTU. òCTKCVKQPU QP VJKU UEJGOG KPENWFG VJG CFFKVKQP QH JKIJN[ TQCUVGF OCNV VQ RTQFWEG FCTM DGGTUþ WUG QH YJGCV HQT YJGCV DGGTUþ CFFKVKQP QH NGUU GZRGPUKXG UVCTEJ UQWTEGU UWEJ CU OCK\G QT TKEGþ QT WUG QH DCEVGTKCN HGTOGPVCVKQP CU RCTV QH VJG RTQEÿ GUU üG.I. $GNIKCP lambic DGGTú. 6JG HGTOGPVCVKQP RTQEGUU KU WUWCNN[ ECTTKGF QWV D[ QPG QH VYQ V[RGU QH DTGYKPI [GCUVþ UQÿECNNGF ÉVQRÿHGTOGPVKPIÌ QT ÉDQVVQOÿHGTOGPVKPIÌ. #NGU CTG VTCFKVKQPCNN[ HGTOGPVGF YKVJ VQRÿHGTOGPVKPI [GCUVU KP HNCVþ QRGP XGUUGNU. 6JGUG [GCUVU CTG EJCTÿ CEVGTKUGF D[ CFJGTGPEG VQ ECTDQP FKQZKFG DWDDNGU CPF YKNN TKUG VQ VJG UWTHCEG CV VJG GPF QH HGTOGPVCVKQP. 6JG [GCUV KU JCTXGUVGF HTQO JGTG HQT WUG KP EQPUGEWVKXG HGTÿ OGPVCVKQPU. #V VJG QVJGT GZVTGOGþ NCIGT DGGT üCPF QVJGT ÉDQVVQOÌú HGTOGPVCVKQPU CTG WUWCNN[ ECTTKGF QWV KP VCNN E[NKPFTQEQPKECN XGUUGNUþ EQPVCKPKPI WR VQ JCNH C OKNÿ NKQP NKVTGUþ WUKPI DQVVQOÿHGTOGPVKPI [GCUVU. #V VJG GPF QH HGTOGPVCVKQPþ VJG [GCUV UGFKOGPVU VQ VJG DQVVQOþ HTQO YJGTG KV KU EQNNGEVGF. +P VJG RTQEGUU QH NCIGT DGGT RTQFWEVKQPþ VJG DTGYKPI [GCUV KPHNWGPEGU VJG qWCNKV[ QH VJG HKPCN RTQFWEV CV VYQ UVCIGUþ PCOGN[ VJG RTKOCT[ CPF UGEQPFCT[ HGTOGPVCÿ VKQPU. 2TQRCICVGF [GCUV KU CFFGF VQ VJG EQQNGF YQTVþ CPF VJG RTKOCT[ HGTOGPVCVKQP RTQEGGFU CV 8ÿ15¸ % HQT 5ÿ1ý FC[U. &WTKPI VJKU RGTKQFþ CNEQJQNþ ECTDQP FKQZKFGþ CPF OWEJ QH VJG CTQOC CTG HQTOGF. 0GZVþ VJG UGEQPFCT[ HGTOGPVCVKQP QT ÉNCIGTKPIÌ HQNNQYUþ C OCVWTCVKQP RGTKQF QH 1ÿ3 YGGMU CV C UNKIJVN[ NQYGT VGORGTCVWTGþ YJKEJ KU PGGFGF VQ QDVCKP VJG RTQRGT DCNCPEG QH HNCXQWTU. &WTKPI VJGUG RTQEGUUGU OCP[ HCEÿ VQTU KPHNWGPEG VJG EJCTCEVGTKUVKEU QH VJG HKPCN DGGTþ QPG DGKPI VJG IGPQV[RG QH VJG NCIGT [GCUV XCTKGV[ WUGFþ UKPEG OCP[ KORQTVCPV DGGT HNCXQWT EQORQPGPVU UWEJ CU CNEQJQNUþ CNFGJ[FGUþ GUVGTU CPF VJKQNU CTG [GCUV OGVCDQNKVGU. 6JKU HQTOU VJG DCUKU HQT TGUGCTEJ KPVQ RQUUKDNG CNVGTCVKQPU QH VJG PCVWTG CPF RGTHQTOCPEG QH VJG [GCUV. ;GCUV EJCTCEVGTKUVKEU UWEJ CU WRVCMG QH ECTDQJ[FTCVGUþ COKPQ CEKFU CPF KQPUþ RTKÿ OCT[ OGVCDQNKUOþ K.G. CDKNKV[ VQ VWTP ECTDQJ[FTCVG KPVQ GPGTI[ CPF CNEQJQNþ UGEQPÿ FCT[ OGVCDQNKUOþ NGCFKPI VQ VJG HQTOCVKQP QH CTQOC EQORQWPFUþ UGFKOGPVCVKQP EJCTCEVGTKUVKEU CPF UVTGUU TGURQPUGþ XKVCNKV[ CPF XKCDKNKV[ CTG CNN CTGCU QH KPVGTGUV HQT VJG DTGYGT CPF DTGGFGT. +P VJKU TGXKGYþ YG YKNN KPVTQFWEG VJG IGPGVKE UVTWEVWTG QH DTGYKPI [GCUVU YKVJ C UVTQPI GORJCUKU QP NCIGT DTGYKPI [GCUV üÉDTGYGT•U [GCUVÌú. 9G YKNN CNUQ FKUEWUU VJG OGVJQFU VJCV CTG CXCKNCDNG HQT DTGGFKPI VJGUG [GCUVU CPF VJG UGNGEVGF CTGCU YJGTG VJGUG DTGGFKPI OGVJQFU JCXG DGGP CRRNKGF VQ KORTQXG VJG RGTHQTOCPEG QH DTGYKPI [GCUVU. ñGEGPV TGXKGYU KP VJG HKGNF KPENWFG -KGNNCPFÿ $TCPFV GV CN. ü1û8ûþ 1ûû5úþ *KPEJNKHHG ü1ûû1úþ 2GPVVKNk CPF ©PCTK ü1ûû1úþ #DDQVV GV CN. ü1ûû3úþ -KGNNCPFÿ$TCPFV ü1ûû4Cþ Dúþ *COOQPF ü1ûû5úþ *CPUGP CPF -KGNNCPFÿ $TCPFV ü1ûû÷Eúþ &GqWKP üôýý1ú CPF 2QNCKPC üôýýôú.
5.2 $TGYGT•U [GCUV: C EJKOGTC KP UGTXKEG 6JG HKTUV RWTGÿEWNVWTGF NCIGT [GCUV YCU KPKVKCNN[ ECNNGF É$QVVQO (GTOGPVKPI SVTCKP ð +Ìþ DWV NCVGT PCOGF Saccharomyces carlsbergensis ü*CPUGP 1ûý8ú. +V KU MGRV CU VJG S. carlsbergensis V[RG UVTCKP %$S1513þ CPF RTQDCDN[ OQUV NCIGT DTGYKPI
14÷
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
[GCUVU WUGF VQFC[ CTG ENQUGN[ TGNCVGF VQ VJKU UVTCKP. #NG DTGYKPI RTGFCVGU NCIGT DTGYKPIþ CPF EQPUGqWGPVN[ CNG [GCUVU FKURNC[ C YKFGT TCPIG QH IGPQOG UVTWEVWTGU ü2GFGTUGP 1û85þ 1ûû4ú üUGG CNUQ %JCRVGT ôú. &GURKVG VJKU XCTKCVKQPþ VJG[ CTG PGXGTÿ VJGNGUU CNN TCVJGT ENQUGN[ TGNCVGF VQ UVCPFCTF NCDQTCVQT[ UVTCKPU QH S. cerevisiae ü2GFGTUGP 1û8÷Cú. 9JKNG QTKIKPCNN[ KPENWFGF KP VJG VCZQP Saccharomyces cerevisiae ü;CTTQY 1û84úþ S. carlsbergensis KU PQY TGEQIPKUGF CU RCTV QH VJG S. pastorianus ITQWR üòCWIJCPÿ/CTVKPK CPF /CTVKPK 1û87ú. #U YKNN DG FGUETKDGF DGNQYþ NCIGT DTGYKPI [GCUVU CTG CNNQRNQKF YKVJ C IGPGVKE UVTWEVWTG TGOKPKUEGPV QH URGEKGU J[DTKFU KP JKIJGT RNCPVU. (QT RTCEVKECN TGCUQPUþ YG UJCNN TGHGT VQ VJGUG [GCUVU D[ VJGKT QTKIKPCN URGEKGU PCOG Saccharomyces carlsbergensis. S. carlsbergensis [GCUVU FKHHGT KP UGXGTCN YC[U HTQO S. cerevisiae. #NVJQWIJ VJG[ CTG PQPÿOCVGTUþ VJG[ FQ PQV GCUKN[ URQTWNCVGþ CPF QPN[ C NQY RGTEGPVCIG QH VJG URQTGU CTG XKCDNG üUGG G.I. )jGTOCPUGP CPF SKIUICCTF 1û81ú. (WTVJGTOQTGþ S. carlsbergensis UGGOU VQ RTGHGT C NQYGT VGORGTCVWTG HQT ITQYVJ VJCP S. cerevisiae. #U OGPVKQPGF CDQXGþ S. carlsbergensis CRRGCTU VQ DG CP CNNQRNQKF üURGEKGU J[DTKFúþ CPF VJKU CEEQWPVU HQT VJG TCVJGT EQORNGZ IGPQOKE UVTWEVWTG QH VJKU [GCUV. 6JG PWÿ ENGCT IGPQOG JCU DGGP UVWFKGF GZVGPUKXGN[þ KPKVKCNN[ D[ VTCPUHGT QH UKPING EJTQOQÿ UQOGU HTQO S. carlsbergensis VQ IGPGVKECNN[ YGNNÿFGHKPGF UVTCKPU QH S. cerevisiaeþ CHVGT YJKEJ VJGUG EJTQOQUQOGU YGTG IGPGVKECNN[ EJCTCEVGTKUGF. 6JWUþ VJG VTCPUHGT QH EJTQOQUQOGU +++ ü0KNUUQPÿ6KNNITGP GV CN. 1û81úþ ò ü0KNUUQPÿ6KNNITGP GV CN. 1û8÷úþ ò++ ü0KNUUQPÿ6KNNITGPþ WPRWDNKUJGFúþ ï ü%CUG[ 1û8÷Dúþ ï++ CPF ï+++ ü2GVGTÿ UGP GV CN. 1û87ú HTQO S. carlsbergensis VQ S. cerevisiae YCU CEEQORNKUJGF. +P UQOG ECUGUþ UWDUVKVWVKQP UVTCKPU EQWNF DG ETGCVGFþ K.G. UVTCKPU VJCV JCF NQUV VJG QTKIKPCN S. cerevisiae EJTQOQUQOGþ OGCPKPI VJCV KP VJGUG ECUGU VJG VTCPUHGTTGF EJTQOQUQOG YCU C HWPEVKQPCN GqWKXCNGPV QH KVU S. cerevisiae EQWPVGTRCTV. (QNNQYKPI VJKUþ OGKQVKE ETQUUKPIÿQXGT DGVYGGP VJG FKHHGTGPV EJTQOQUQOGU YCU CUUC[GF KP FKRNQKF ETQUUGU QH VJG UWDUVKVWVKQP UVTCKPU YKVJ UVCPFCTF S. cerevisiae UVTCKPU. 6JG EJTQOQUQOGU FGTKXGF HTQO S. carlsbergensis YGTG HQWPF VQ DG QH VJTGG MKPFU: Kú JQOQNQIQWU EJTQOQUQOGU VJCV YQWNF TGEQODKPG PQTOCNN[ YKVJ VJGKT S. cerevisiae EQWPVGTÿ RCTVUþ KKú JQOQGQNQIQWU EJTQOQUQOGU VJCV TCTGN[ TGEQODKPGF YKVJ VJGKT S. cerevisiae EQWPVGTRCTVUþ CPF KKKú OQUCKE EJTQOQUQOGU VJCV YGTG EQORQUGF QH JQOQNQÿ IQWU CPF JQOQGQNQIQWU UGIOGPVU. 6JG GOGTIKPI RKEVWTG QH C J[DTKF QTICPKUO JCU DGGP GZVGPFGF D[ SQWVJGTP J[ÿ DTKFKUCVKQP QH IGPQOKE &0# HTQO S. carlsbergensis QT CFFKVKQP CPF UWDUVKVWVKQP UVTCKPUþ GORNQ[KPI &0# RTQDGU QTKIKPCVKPI HTQO ENQPGF S. cerevisiae IGPGU QT S. carlsbergensis IGPGU: BAP2 üEJT. ++ú ü-QFCOC GV CN. ôýý1úþ HIS4 üEJT. +++ú ü0KNUÿ UQPÿ6KNNITGP GV CN. 1û81úþ LEU2 üEJT. +++ú ü2GFGTUGP 1û85úþ MAT üEJT. +++úþ HML üEJT. +++ú, HMR üEJT. +++úþ CPF SUP-RL1 üEJT. +++ú ü*QNODGTI 1û8ôúþ ILV1 üEJT. òúþ CAN1 üEJT. òúþ CYC7 üEJT. òúþ CPF URA3 üEJT. òú ü0KNUUQPÿ6KNNITGP GV CN. 1û8÷úþ MXR1 üEJT. òú ü*CPUGP 1ûûûúþ ILV3 üEJT. ïú CPF CYC1 üEJT. ïú ü%CUG[ 1û8÷CþDúþ ILV5 üEJT. ï++ú CPF ILV2 üEJT. ï+++ú ü2GVGTUGP GV CN. 1û87úþ MET2 üEJT. ï+òú ü*CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû4úþ CPF ATF1 ü(WjKK GV CN. 1ûû4ú. ùUWCNN[þ VYQ FKXGTIGPV V[RGU QH GCEJ IGPG KP qWGUVKQP JCXG DGGP HQWPFþ QH YJKEJ QPG KPXCTKCDN[ GZJKDKVGF C TGUVTKEVKQP CPF J[DTKFKUCVKQP RCVVGTP KFGPVKECN QT CNOQUV KFGPVKECN VQ VJCV HQWPF KP S. cerevisiaeþ YJKNG VJG QVJGT UJQYGF FKXGTIGPV RCVVGTPU. 6JG HQTOGT KU QHVGP TGHGTTGF VQ CU VJG S. cerevisiaeÿNKMG V[RG üXCTKQWUN[ FGUETKDGF CU VJG SEÿþ EGTÿ
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
147
QT ÿCE V[RGú CPF VJG QVJGT VJG S. pastorianus üSRÿúþ NCIGT üLIÿúþ QT S. carlsbergensisÿURGEKHKE üÿCAú V[RG. 9G YKNN WUG VJG FGPQVCVKQPU ÿCE CPF ÿCA. 6JG J[DTKF PCÿ VWTG QH S. carlsbergensis JCU TGEGPVN[ DGGP TGÿEQPHKTOGF KP UQOG NCTIG UVWFKGU GOÿ RNQ[KPI J[DTKFKUCVKQP YKVJ TCFKQCEVKXG RTQDGU VQ RWNUGFÿHKGNF GNGEVTQRJQTGUKUÿ UGRCTCVGF EJTQOQUQOGU ü6COCK GV CN. 1ûû8î ;COCIKUJK CPF õICVC 1ûûûú CU JCU VJG OQUCKE UVTWEVWTG QH UGXGTCN EJTQOQUQOGUþ D[ IGPQOKE &0# J[DTKFKUCVKQP VQ S. cerevisiae IGPG CTTC[U ü-QFCOC GV CN. &KXGTUKV[ QH EJTQOQUQOCN UVTWEVWTG KP NCIGT DTGYKPI [GCUVþ manuscript in preparationú. 6JG PWENGQVKFG UGqWGPEGU QH UGXGTCN S. carlsbergensisÿURGEKHKE IGPGU JCXG DGGP FGVGTOKPGF CPF EQORCTGF VQ VJG GqWKXCNGPV PWENGQVKFG UGqWGPEGU HTQO S. cerevisiae. +PXCTKCDN[þ õñ( UGqWGPEGU QH ÿCE CPF ÿCA IGPGU JCXG DGGP HQWPF VQ DG JKIJN[ TGNCVGF CPF CNYC[U QH VJG UCOG NGPIVJþ YJKNG PQPÿEQFKPI UGqWGPEGU UJQY OWEJ NGUU IGPGTCN KFGPVKV[. 6JWUþ VJG ILV1-CA õñ( CPF VJG S. cerevisiae ILV1 IGPGU CTG 8÷ó KFGPVKECNþ YJGTGCU VJG FGFWEGF RQN[RGRVKFG UGqWGPEGU CTG û÷ó KFGPVKECN ü)jGTOCPUGP 1ûû1ú. 6JG UCOG EQORCTKUQPU QH VJG ILV2 ü)jGTOCPUGP 1ûû1úþ MET2 ü*CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû4úþ MET10 ü*CPUGP GV CN. 1ûû4úþ ACB1 ü$øTUVKPI GV CN. 1ûû7úþ HO ü6COCK GV CN. ôýýýúþ MET14 ü,QJCPPGUGP CPF *CPUGP ôýýôúþ MXR1 ü*CPUGP GV CN. ôýýôúþ BAP2 ü-QFCOC GV CN. ôýý1úþ CPF ATF1 ü(WjKK GV CN. 1ûû÷ú IGPGU IKXG PWODGTU QH 78ÿ88ó üPVú CPF 7÷ÿû7ó üCCúþ TGURGEÿ VKXGN[. SGXGTCN SQWVJGTP J[DTKFKUCVKQP GZRGTKOGPVU JCXG KPFKECVGF VJCV VJG S. carlsbergensis -CE V[RG QH CP[ IKXGP IGPG KU KFGPVKECN QT RTCEVKECNN[ KFGPVKECN VQ VJG EQTTGURQPFKPI S. cerevisiae IGPG ü0KNUUQPÿ6KNNITGP GV CN. 1û8÷î 2GVGTUGP GV CN. 1û87ú. 6JKU PQVKQP JCU DGGP EQPHKTOGF D[ RCTVKCN QT HWNN UGqWGPEKPI QH C HGY S. carlsbergensis -CE IGPGU ü*CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû4î (WjKK GV CN. 1ûû÷î $øTUVKPI GV CN. 1ûû7î ,QJCPPGUGP CPF *CPUGP ôýýôú. S. carlsbergensis UGGOU VQ DG VJG TGUWNV QH J[DTKFKUCVKQP DGVYGGP S. cerevisiae CPF CPQVJGT Saccharomyces [GCUV. SKPEG VJG QDUGTXGF FKXGTIGPEG KU EQTTGNCVGF YKVJ C FTQR KP OGKQVKE TGEQODKPCVKQP GZRGEVGF VQ ECWUG HGTVKNKV[ DCTTKGTUþ VJKU J[ÿ RQVJGVKECN J[DTKFKUCVKQP OWUV DG EQPUKFGTGF CU DGKPI KPVGTURGEKHKE. òCTKQWU ECPFKÿ FCVGU HQT VJG PQPÿS. cerevisiae RCTGPV JCXG DGGP UWIIGUVGF. õPG YCU CPQVJGT [GCUV KUQNCVGF D[ *CPUGP ü1ûý8úþ É$QVVQO (GTOGPVKPI ;GCUV ð++Ìþ KPKVKCNN[ PCOGF Saccharomyces monacensis ü%$S 15ý3ú. SQWVJGTP J[DTKFKUCVKQP CPF ENQPKPI GZRGTKÿ OGPVU KPFKECVGF VJCV QPN[ S. carlsbergensisÿURGEKHKE üÿCAú IGPGU GZKUVGF KP VJKU [GCUV ü2GFGTUGP 1û8÷Cî Dî *CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû4î $øTUVKPI GV CN. 1ûû7ú. *QYGXGTþ KV JCU TGEGPVN[ DGGP UJQYP VJCV CNVJQWIJ FGHKEKGPV KP VJG -CE XGTÿ UKQP QH OCP[ IGPGUþ UVTCKP %$S15ý3 WPFQWDVGFN[ EQPVCKPU S. cerevisiaeÿNKMG IGÿ PQOKE &0# ü#PFGTUGP GV CN. ôýýýî %CUCTGIQNC GV CN. ôýý1î -QFCOC GV CN. &KXGTÿ UKV[ QH EJTQOQUQOCN UVTWEVWTG KP NCIGT DTGYKPI [GCUVþ manuscript in preparationúþ C PQVKQP VJCV JCU DGGP UWRRQTVGF D[ #(L2 üHNWQTGUEGPV CORNKHKGF HTCIOGPV NGPIVJ RQN[OQTRJKUOUú UVWFKGU üFG $CTTQU LQRGU GV CN. ôýýôú. #PQVJGT UWIIGUVGF ECPFKÿ FCVG HQT VJG RCTGPV UJKR YCU Saccharomyces bayanus ü%$S 38ýú. 6JKU J[RQVJGUKU YCU QTKIKPCNN[ DCUGF QP &0#ÿ&0# TGCUUQEKCVKQP GZRGTKOGPVU üòCWIJCPÿ/CTVKPK CPF -WTV\OCP 1û85úþ CPF TGEGPVN[ ICKPGF UWRRQTV D[ HKPFKPIU QH KFGPVKECN PQPÿS. cerevisiaeÿNKMG UGqWGPEGU KP UVTCKPU QH S. carlsbergensis CPF KP VJG S. bayanus %$S 38ý V[RG UVTCKP ü6COCK GV CN. 1ûû8î ;COCIKUJK CPF õICVC 1ûûûî -QFCOC GV CN. ôýý1î %CUCTGIQNC GV CN. ôýý1ú. *QYGXGTþ UVTCKP %$S 38ý KP KVUGNH CRRGCTU VQ DG C
148
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
J[DTKF [GCUVþ YJKEJ EQPVCKPU VYQ XGTUKQPU QH OCP[ IGPGUþ QH YJKEJ QPG KU KFGPVKÿ ECN VQ VJG ÿ%# V[RGU QH S. carlsbergensis CPF QPG KU CDQWV 7óþ FKXGTIGF ü2GFGTUGP 1û8÷Cî *CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû4î %CUCTGIQNC GV CN. ôýý1ú. 6JG OQUV QDÿ XKQWU J[RQVJGUKU VQ GZRNCKP CNN QH VJGUG QDUGTXCVKQPU KU VJCV S. carlsbergensis/S. pastorianus TGRTGUGPVU QPG J[DTKFKUCVKQP GXGPV CPF S. bayanus %$S 38ý CPQVJGT. 6JG[ JCXG KP EQOOQP VJG IGPQOKE UGqWGPEGU TGRTGUGPVGF D[ VJG ÿCA IGPGUþ YJKEJ UVKNN JCXG PQV DGGP HQWPF CNQPG KP CP[ [GCUV URGEKGU. ñGEGPV GZRGTKOGPVCÿ VKQP CV õUCMC ùPKXGTUKV[ KPFKECVGU VJCV VJG S. bayanus UVTCKPU +(õ53û CPF +(õ1û48 OKIJV DG ÉRWTGÌ EGNN NKPGUþ VJG QPN[ IGPQOKE EQPVGPV QH YJKEJ EQTTGURQPFU VQ VJG PQPÿS. cerevisiaeÿNKMG IGPQOG QH S. carlsbergensis DTGYKPI [GCUV üñCKPKGTK GV CN. OCPWUETKRV KP RTGRCTCVKQPú. (QT VJG RWTRQUG QH DTGGFKPI DTGYGT•U [GCUV VJG qWGUVKQP QH VJG EQR[ PWODGT QH GCEJ XGTUKQP QH C IKXGP EJTQOQUQOG QT EJTQOQUQOCN TGIKQP KU jWUV CU KORQTVCPV CU VJG qWGUVKQP QH VJG J[DTKF PCVWTG üUGG CNUQ %JCRVGT ôú. 6JKU qWGUVKQP JCU DGGP CRRTQCEJGF HQT VJG ILV2 NQEWU KP C S. carlsbergensis NCIGT RTQFWEVKQP UVTCKP ü)jGTÿ OCPUGP GV CN. 1û88î -KGNNCPFÿ$TCPFV GV CN. 1û8ûú. #U RCTV QH C UVWF[ QH VJG EQPUGÿ qWGPEGU QH GNKOKPCVKPI ILV2 IGPG HWPEVKQP KP NCIGT DTGYKPI [GCUVþ FGNGVKQPU QH VJG VYQ YKNF V[RG CNNGNGU YGTG EQPUVTWEVGF in vitro. ùUKPI VJG VYQÿUVGR IGPG TGÿ RNCEGOGPV VGEJPKqWG üSEJGTGT CPF &CXKU 1û7ûú CPF VJG DCEVGTKCN )418 üIGPGVKEKPú TGUKUVCPEG OCTMGT RTQXKFGF YKVJ C [GCUV RTQOQVGT ü;QEWO 1û85úþ [GCUVU YGTG QDÿ VCKPGF YJKEJ ECTTKGF QPG QT VJG QVJGT FGNGVKQP CNNGNG QH ILV2 KPUVGCF QH VJG YKNF V[RG CNNGNG. SQWVJGTP CPCN[UKU QH VJGUG UVTCKPU UJQYGF VJCV VJG RCTVKEWNCT NCIGT DTGYKPI [GCUV EQPVCKPU VYQ EQRKGU QH GCEJ QH VJG VYQ XGTUKQPU QH VJG ILV2 IGPG. )GPGTCNKUCVKQP QH VJKU HKPFKPI YQWNF OGCP VJCV NCIGT DTGYKPI [GCUV KU CNÿ NQVGVTCRNQKFþ DWV TGEGPV UVWFKGU UWIIGUV C RGPVCUQOKE EQPVGPV QH VJG HIS4ÿ EQPVCKPKPI EJTQOQUQOG +++ TGIKQPU ü*QHHOCPP ôýýýú. (WTVJGTþ S. carlsbergensis EJTQOQUQOGU QH VJTGG UK\GU J[DTKFKUG YKVJ C EJTQOQUQOG ï RTQDG ü%CUG[ 1û8÷Dú QT YKVJ C EJTQOQUQOG +++ RTQDG ü2GFGTUGP 1û8÷Dúþ CPF TGEGPVN[ CPQVJGT NCIGT DTGYKPI UVTCKP QH S. carlsbergensis YCU HQWPF PQV VQ EQPVCKP C PQPÿS. cerevisiaeÿ NKMG XGTUKQP QH VJG ATF2 IGPG ü;QUJKOQVQ GV CN. 1ûûûú. 6JWUþ GXGP VJQWIJ S. carlsbergensis NCIGT [GCUV JCU C VQVCN &0# EQPVGPV CRRTQZKOCVGN[ EQTTGURQPFKPI VQ VGVTCRNQKF[ ü*QHHOCPP ôýýýúþ KV CRRGCTU VQ DG KTTGIWNCT KP KVU EJTQOQUQOG UGVÿ WR CPF RTQDCDN[ CPGWRNQKF HQT UQOG EJTQOQUQOGUþ QT EJTQOQUQOG TGIKQPU. $GCTKPI KP OKPF VJG IGPQOKE EQORNGZKV[ QH S. carlsbergensis DTGYGT•U [GCUVþ C HWTVJGT qWGUVKQP CTKUGU: #TG VJG VYQ XGTUKQPU QH CP[ IKXGP IGPG GZRTGUUGF GqWCNN[ QT FKHHGTGPVKCNN[! &Qþ G.I.þ VTCPUETKRVKQP HCEVQTU QPN[ TGEQIPKUG VJGKT JQOQNQIQWU EQIPCVG UGqWGPEGU QT FQ VJG[ CNUQ TGEQIPKUG VJGKT JQOQGQNQIQWU EQWPVGTRCTVU! %QWNF UOCNN FKHHGTGPEGU KP VJGKT VCTIGV CHHKPKVKGU CHHGEV ÉFQYPUVTGCOÌ EGNNWNCT DKQÿ EJGOKUVT[ CPF VJWU HGTOGPVCVKQP RGTHQTOCPEG! +P QVJGT YQTFU: *QY FQ VJG JQÿ OQGQNQIQWU IGPQOG RCTVU EQÿQRGTCVG KP VGTOU QH IGPG GZRTGUUKQP! &WG VQ VJG NCEM QH NCTIGÿUECNG IGPQOKE UGqWGPEG FCVC HQT S. carlsbergensisþ VJGTG JCXG DGGP PQ INQDCN UVWFKGU QH VTCPUETKRVKQP QH VJG S. carlsbergensisÿURGEKHKE IGPGU. *QYGXGTþ 0QTVJGTP CPCN[UKU QH VJG BAP2 JQOQGQNQIWGU UVTQPIN[ KPFKECVGF FKHHGTGPVKCN GZÿ RTGUUKQP QH VJGUG ü-QFCOC GV CN. ôýý1úþ CPF TGXGTUG VTCPUETKRVKQPÿEQWRNGF 2%ñ UJQYGF FKHHGTGPVKCN FGTGRTGUUKQP QH VJG UGVU QH MET2 CPF MET14 JQOQGQNQIWGU ü,QJCPPGUGP CPF *CPUGP ôýýôú. SWEJ QDUGTXCVKQPU OC[ DG UGGP CU KPFKECVKQPU VJCV
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
14û
VJKU DTGYKPI [GCUV KU KP VJG CEVWCN RTQEGUU QH URGEKCVKQPþ YJKEJ OC[ TGUWNV KP UQOG IGPGU DGEQOKPI TGFWPFCPV CPF UKNGPEGFþ CPF QVJGTU ICKPKPI PGY HWPEVKQPU. 6Q UWOOCTKUGþ S. carlsbergensis DTGYGT•U [GCUV CRRGCTU VQ DG C NCTIGN[ CNÿ NQVGVTCRNQKF URGEKGU J[DTKFþ YJKEJ EQPVCKPU RCTVU QH VYQ FKXGTIGF IGPQOGUþ QPG FGTKXGF HTQO S. cerevisiae CPF QPG YJKEJ KU FGTKXGF HTQO CP WPMPQYP CPEGUVQT. SGZWCN TGRTQFWEVKQP KP VJKU QTICPKUO KU KORCKTGFþ RGTJCRU FWG VQ VJG OQUCKE UVTWEÿ VWTG QH UQOG EJTQOQUQOGUþ RGTJCRU FWG VQ OWVCVKQPU KP IGPGU KORQTVCPV HQT URQTWNCVKQPþ GVE. %NCUUKECN IGPGVKE CPCN[UKU YKVJ VGVTCFU KU VJGTGHQTG PQV RQUUKDNGþ C HCEV YKVJ NQPI TGEQIPKUGF ITCXG EQPUGqWGPEGU HQT DTGGFKPI. *QYGXGTþ QPG OC[ URGEWNCVG YJGVJGT VJG NQY GHHKEKGPE[ QH UGZWCN TGRTQFWEVKQP JCU EQPVTKDWVGF VQ OCKPVCKPKPI C TGNCVKXGN[ JKIJ IGPGVKE UVCDKNKV[ QH C [GCUV RQRWNCVKQP YKVJ UCVKUHCEÿ VQT[ EJCTCEVGTKUVKEU.
5.3 *QY VQ DTGGF DTGYGT•U [GCUV SGNGEVKQP HQT VJG [GCUV EGNNU DGUV CFCRVGF VQ HGTOGPVCVKQP EQPFKVKQPU FWTKPI JKUVQT[ JCU WPFQWDVGFN[ TGUWNVGF KP IGPGVKE EJCPIGUþ YJKEJ RTQXKFGF VJG DTGYKPI [GCUV YKVJ KVU DCUKE CDKNKVKGU VQ RTQFWEG DGGT. *QYGXGTþ VQ HWTVJGT KORTQXG [GCUV EJCTCEÿ VGTKUVKEUþ FKTGEVGF DTGGFKPI KU PGEGUUCT[. $TGGFKPI CPF KORTQXGOGPV QH CP[ QTICPÿ KUO OCKPN[ TGNKGU QP VJTGG RTKPEKRNGU: ETQUUÿJ[DTKFKUCVKQPþ OWVCIGPGUKUþ QT IGPGVKE VTCPUHQTOCVKQP üUGG %JCRVGT ôú. 9JKNG VJG HQTOGT VYQ OGVJQFU JCXG DGGP GOÿ RNQ[GF HQT EGPVWTKGUþ IGPGVKE VTCPUHQTOCVKQP KU QPN[ PQY DGKPI RGTHGEVGF CU C VGEJÿ PQNQI[þ DWV CNN ECP DG WUGF VQ DTGGF DTGYGT•U [GCUV. +P URKVG QH VJG IGPGVKECNN[ EQORNKECVGF PCVWTG QH S. carlsbergensis NCIGT [GCUVþ ENCUUKECN DTGGFKPI KU RQUUKDNG. 6JWUþ VJG KUQNCVKQP QH NCIGT [GCUV URQTG UGITGICPVU YKVJ VJG CDKNKV[ VQ OCVG JCU DGGP FGUETKDGF ü)jGTOCPUGP CPF SKIUICCTF 1û81ú. SWEJ EGNN NKPGU YGTG HQWPF VQ DG CNNQRNQKF CPF WUWCNN[ FKRNQKF ü0KNUUQPÿ6KNNITGP GV CN. 1û8÷î 2GVGTUGP GV CN. 1û87î *CPUGP CPF -KGNNCPFÿ$TCPFVþ 1ûû÷Cî 1ûû÷Dî ,QJCPÿ PGUGP CPF *CPUGP ôýýôî *CPUGP GV CN. ôýýôúþ CNVJQWIJ CPGWRNQKF[ JCU CNUQ DGGP FGVGEVGF ü*QHHOCP ôýýýú. 6JGKT EQORNKECVGF IGPGVKE UVTWEVWTG KU PQV UWTRTKUKPIþ CU VJG[ CTG FGTKXGF HTQO CP KTTGIWNCT CNNQVGVTCRNQKFþ DWV VJKU OC[ CEVWCNN[ DG DGPGHKÿ EKCN HQT DTGGFKPI RWTRQUGU. *[DTKFU HQTOGF HTQO CP CTTC[ QH UWEJ OCVGTU HQTO C qWKVG JGVGTQIGPGQWU RQRWNCVKQP QH RQVGPVKCN DTGYKPI UVTCKPU ü)jGTOCPUGP CPF SKIUICCTF 1û81úþ UQOG QH YJKEJ EQWNF DG DGVVGT UWKVGF VQ PGY DTGYKPI EQPFKVKQPU. 6JG WUG QH OCVGTU HTQO QVJGT [GCUVU ü$KNKPUMK GV CN. 1û87î $KNKPUMK CPF %CUG[ 1û8ûú EQWNF HWTVJGT CNNQY VJG KPVTQFWEVKQP QH FGUKTGF EJCTCEVGTU HTQO C PQPÿ DTGYKPI [GCUV KPVQ YGNN RGTHQTOKPI DTGYKPI [GCUVU. (WTVJGTþ TGEGUUKXG OWVCPVU QH VJG CNNQFKRNQKF EGNN NKPGU ECP DG QDVCKPGF ü)jGTOCPUGP 1û83úþ UQOGVJKPI VJCV KU PQV RQUUKDNG YKVJ VJG RCTGPVCN CNNQVGVTCRNQKFU. ùRQP ETQUUKPIþ C HGY QH VJGUG OCVKPIÿ RTQHKEKGPV UVTCKPU HQTOGF J[DTKFU VJCV RGTHQTOGF CNOQUV NKMG VJG RCTGPVCN NCIGT [GCUV KP RKNQV HGTOGPVCVKQP GZRGTKOGPVU ü)jGTOCPUGP CPF SKIUICCTF 1û81ú. SWEJ RCKTU QH OCVGTU CTG QH ITGCV XCNWG HQT VCTIGVGF DTGGFKPI RTQjGEVUþ CU WUWCNN[ QPN[ QPG EQR[ QH GCEJ V[RG QH CP[ IKXGP IGPG KU RTGUGPV. õVJGT OGVJQFU HQT DTGGFKPI QH NCIGT [GCUV YJKEJ FQ PQV TGN[ QP IGPGVKE VTCPUHQTOCVKQP KPENWFG ÉTCTG OCVKPIÌ QH
15ý
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
EJCTCEVGTKUGF JCRNQKF [GCUVU YKVJ DTGYKPI [GCUVU ü6WDD GV CN. 1û81úþ CPF RTQVQRNCUV HWUKQPþ CXQKFKPI VJG qWGUVKQP QH VJG OCVKPI V[RG CNVQIGVJGT üñWUUGNN CPF SVGYCTV 1û7ûî ùTCPQ GV CN. 1ûû3CþDú. 6JG VGEJPKqWGU FGUETKDGF UQ HCT KORNKECVG VJG TCVJGT WPRTGFKEVCDNG VTCPUHGT QH OCP[ IGPGUþ CPF YJKNG VJKU OC[ DG CP CFXCPVCIG KP UQOG ECUGUþ C ITGCVGT FGITGG QH RTGEKUKQP KU WUWCNN[ PGGFGF KP FKTGEVGF DTGGFKPI RTQjGEVU. /QFGTP TGEQODKPCPV IGPG VGEJPQNQI[ RTQXKFGU VJG [GCUV DTGGFGT YKVJ VJG OGCPU VQ KPCEVKXCVG QT CFF RCTVKEWNCT IGPGU VQ VJG DTGYKPI [GCUV. õTKIKPCNN[þ IGPGVKE VTCPUHQTOCVKQP QH DTGYÿ KPI [GCUV TGNKGF QP VJG WUG QH ôµOÿDCUGF RNCUOKFU ü)jGTOCPUGP 1û83î *GPFGTUQP GV CN. 1û85ú. 6JGUG RNCUOKFU TGUWNV KP JKIJ GZRTGUUKQP FWG VQ C JKIJ EQR[ PWODGTþ DWV EGNNWNCT RNCUOKF KPJGTKVCPEG KU KPEQORNGVG FWG VQ OKVQVKE NQUU üUGG G.I. ñQÿ OCPQU GV CN. 1ûûôú. +PVGITCVKQP QH &0# UGqWGPEGU KPVQ VJG DTGYKPI [GCUV IGPQOGþ QP VJG QVJGT JCPFþ TGUWNVU KP C TCVJGT UVCDNG IGPGVKE KPJGTKVCPEGþ FGRGPFKPI QP VJG GZCEV OGVJQF GORNQ[GF. 6JGUG OGVJQFU FGRGPF QP TGEQODKPCVKQP DGVYGGP JQÿ OQNQIQWU UGqWGPEGU QP VJG [GCUV EJTQOQUQOG CPF VJG KPVGITCVKQP RNCUOKFþ CU YGNN CU VJG RTGUGPEG QH C UGNGEVKQP OCTMGT. 6JG EJQKEG QH VGEJPKqWG OQUVN[ FGRGPFU QP VTCPUHQTOCDKNKV[ QH VJG [GCUV UVTCKP KP qWGUVKQPþ CPF YJGVJGT KPVTQFWEVKQPþ KPCEVKXCÿ VKQPþ QT CNVGTCVKQP QH C IGPG KU VJG RWTRQUG. #P CFXCPVCIG QH ÉUKORNGÌ KPVGITCVKQP QH C EQORNGVG RNCUOKF EQPVCKPKPI [GCUV UGqWGPEGU KU VJG UKORNKEKV[ QH VJG RTQEGÿ FWTGþ DWV UKPEG CFjQKPKPI JQOQNQIQWU UGqWGPEGU CTG RTGUGPV CHVGT KPVGITCVKQPþ OKÿ VQVKE UVCDKNKV[ KU PQV EQORNGVG üUG G.I. ñQOCPQU GV CN. 1ûûôú. SKPINGÿUVGR KPVGITCVKQP QH C &0# HTCIOGPVþ EQPVCKPKPI [GCUV UGqWGPEGU CV DQVJ VGTOKPK üñQVJUVGKP 1ûû1úþ KU CNUQ UVTCKIJVHQTYCTFþ DWV YKNN WUWCNN[ TGUWNV KP VJG RGTOCPGPV KPUGTVKQP CPF VJWU GZRGPFKVWTG QH C IGPGVKE OCTMGT. 6JG VYQÿUVGR UWDUVKVWVKQP RTQEGFWTGþ QTKIKPCNN[ FGXKUGF D[ SEJGTGT CPF &CXKU ü1û7ûúþ KU OQTG EWODGTUQOGþ DWV QP VJG QVJGT JCPF TGUWNVU KP C RGTOCPGPV CPF UVCDNG IGPGVKE EJCPIGþ CPF NGCXGU PQ TGFWPFCPV &0#. 6JG VGEJPKqWG TGNKGU QP C UKPING KPKVKCN TGEQODKPCVKQP GXGPVþ CPF KV OC[ DG WUGF HQT TGOQXKPI IGPGU QT RCTVU QH IGPGUþ HQT KPVGITCVKQP QH JQOQNQIQWU IGPGU YKVJ CNÿ VGTGF UGqWGPEGþ QT HQT KPVGITCVKQP QH JGVGTQNQIQWU IGPGU üKP VJKU ECUG JQOQNQIQWU [GCUV &0# UGqWGPEGU PGGF VQ GODTCEG VJG JGVGTQNQIQWU IGPGú. *QOQNQIQWU TGÿ EQODKPCVKQP VCMGU RNCEG VJTQWIJ QPG GPF QH VJG IGPG QH KPVGTGUV üQT VJTQWIJ QPG QH VJG HTCIOGPVU GODTCEKPI C JGVGTQNQIQWU IGPG ECUUGVVGúþ VJWU KPVGITCVKPI VJG EQOÿ RNGVG PQPÿTGRNKECVKPI RNCUOKF. # FQOKPCPV UGNGEVKQP OCTMGT KFGPVKHKGU RTQRGT KPVGÿ ITCPVUþ YJKEJ PQY JCXG EQRKGU QH VJG &0# UGqWGPEGU VJTQWIJ YJKEJ TGEQODKPCÿ VKQP VQQM RNCEGþ GODTCEKPI VJG IGPG QT IGPGU QH KPVGTGUV. # UGEQPFCT[ TGEQODKPCVKQP GXGPV FKUVCN VQ VJG RTKOCT[ UKVG QH TGEQODKPCVKQP CPF VQ VJG JGVÿ GTQNQIQWU QT CNVGTGF JQOQNQIQWU IGPG UGqWGPEGU TGUWNVU KP VJG GZEKUKQP QH RNCUOKF UGqWGPEGUþ KPENWFKPI VJG UGNGEVKQP OCTMGT. õPG YGNNÿMPQYP FKUCFXCPVCIG YKVJ VJG VYQÿUVGR RTQEGFWTG WUGF VQ DG VJCV VJG UGCTEJ HQT ÉNQQRÿQWVUÌþ VJG TGUWNVU QH VJG UGEQPFCT[þ TCTG TGEQODKPCVKQP GXGPVþ YCU qWKVG NCDQTKQWU. %QOOQPN[ WUGF EQWPVGTÿUGNGEVKQP HQT URGEKHKE PWVTKGPV TGqWKTGOGPVUþ UWEJ CU ura3 ü$QGMG GV CN. 1û84ú KU PQV UWKVCDNG YJGP RCTGPVCN UVTCKPU CTG RTQVQVTQRJKE. ñGEGPVN[þ JQYGXGTþ VJG KPENWUKQP QH UWKVCDNG EQWPVGTÿUGNGEVKQP OCTMGTU QP KPVGITCVKQP RNCUOKFU JCU DGGP FGUETKDGF ü-CYCJCVC GV CN. 1ûûûî õNGUGP GV CN. ôýýýî #MCFC GV CN. ôýýôú GPCÿ DNKPI GCU[ UGNGEVKQP HQT ÉNQQRÿQWVUÌ. #U NCIGT [GCUV KU RQN[RNQKFþ KV YKNN QHVGP DG PGEGUUCT[ VQ KPVGITCVG C IKXGP IGPG QP VYQ QT OQTG JQOQNQIQWU EJTQOQUQOGUþ KP
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
151
QTFGT VQ CXQKF ÉTGRCKTÌ QH VJG CNVGTCVKQP D[ OKVQVKE TGEQODKPCVKQP. *QYGXGTþ KV KU PQV PGEGUUCT[ VQ KPENWFG KPVGITCVKQP KP VJG FKXGTIGPV EJTQOQUQOGUþ UKPEG VJG HTGÿ qWGPE[ QH OKVQVKE TGEQODKPCVKQP DGVYGGP VJG JQOQGQNQIQWU &0# UGqWGPEGU KU XGT[ NQY üñGUPKEM GV CN. 1û8ûî *QHHOCPP ôýýýú. +P ECUG QPG YCPVU VQ TGOQXG CNN CEVKXKV[ QH C YKNFÿ V[RG [GCUV IGPGþ VJG EQRKGU QP VJG FKXGTIGPV EJTQOQUQOGU OWUV CNUQ DG KPCEVKXCVGF. ñGICTFNGUU QH VJG VTCPUHQTOCVKQP OGVJQFþ IGPGVKE OCTMGTU CTG PGGFGF HQT UGNGEÿ VKQP. #U DTGYKPI CPF QVJGT KPFWUVTKCN [GCUVU FQ PQV EQPVCKP CWZQVTQRJ[ OCTMGTUþ UQÿECNNGF ÉFQOKPCPVÌ üK.G. FQOKPCPV QXGT YKNFÿV[RGú OCTMGTU CTG GZRGFKGPV. ©CTN[ GZRGTKOGPVU GORNQ[GF VJG EQRRGT TGUKUVCPEGÿEQPHGTTKPI CUP1 OCTMGT ü*GPFGTUQP GV CN. 1û85ú. *QYGXGTþ VJG CEqWKTGF TGUKUVCPEG FGRGPFU QP JKIJÿEQR[ PWODGT üôµOú GZRTGUUKQPþ CPF VJGTGHQTG CUP1 KU PQV VJG EJQKEG HQT KPVGITCVKQP. #P QRVKQP HQT VJKU KU VJG SMR1 UWNHQOGVWTQP OGVJ[N üS/úÿTGUKUVCPV ILV2 OWVCPV CNNGNGþ YJKEJ ECWUGU FQOKPCPV S/ TGUKUVCPEG D[ GPEQFKPI C HQTO QH CEGVQJ[FTQZ[ CEKF U[PVJCUG TGUKUVCPV VQ VJG JGTDKEKFG ü(CNEQ CPF &WOCU 1û85î %CUG[ GV CN. 1û88úþ DWV VJG OCTMGT JCU VJG FKUCFXCPVCIG VJCV URQPVCPGQWU TGUKUVCPEG OC[ CTKUG KP [GCUV EGNNU. 6JG AUR1ÿ% OCTMGTþ YJKEJ IKXGU TGUKUVCPEG VQ CWTGQDCUKFKP #þ KU CNUQ DCUGF QP C [GCUV IGPGþ DWV UKPEG AUR1ÿ% KU RCTVN[ U[PVJGVKEþ CRRGCTCPEG QH URQPVCPGQWU TGUKUVCPEG KP [GCUV KU DGNKGXGF VQ DG NGUU QH C RTQDNGO ü*CUJKFCÿõMCFQ GV CN. 1ûû8î *CPUGP GV CN. +PEQTRQTCVKQP QH VJG FQOKPCPV hphþ nat CPF AUR1 IGPG OCTMGTU CPF VJG lac< TGRQTVGT U[UVGO KP VJG ECUUGVVGÿDCUGF R;% RNCUOKF U[UVGO. ùUG KP Saccharomyces cerevisiaeþ Saccharomyces carlsbergensis, CPF Saccharomyces kluyveri. Manuscript in preparationú. +P CP[ GXGPVþ DCEVGTKCN TGUKUVCPEG IGPGU GZÿ RTGUUGF HTQO [GCUV RTQOQVGTU CRRGCT VQ DG VJG DGUV TQWVKPG UGNGEVKQP VQQNU HQT WUG KP KPFWUVTKCN [GCUVUþ CPF WUGF YKVJ VJG VYQÿUVGR UWDUVKVWVKQP RTQEGFWTGþ DCEVGTKCN &0# UGqWGPEGU YKNN GXGPVWCNN[ DG FKURQUGF QH. õPG YKFGN[ WUGF OCTMGT KU VJG IGÿ PGVKEKP ü)418ú TGUKUVCPEG OCTMGT ü;QEWO 1û8÷úþ CPF QVJGT QRVKQPU KPENWFG VJG IN[RJQUCVGþ PQWTUGQVJTKEKPþ J[ITQO[EKPþ CPF CWTGQDCUKFKP OCTMGTU ü-WP\G GV CN. 1û8ûî )QNFUVGKP CPF /E%WUMGT 1ûûûî õNGUGP GV CN. ôýýýú. 6JWUþ C qWKVG FKXGTUG CTTC[ QH OGVJQFU GZKUVU HQT FKTGEVGF IGPGVKE OQFKHKECVKQP QH DTGYKPI [GCUVU. $GNQYþ YG YKNN FGUETKDG [GCUV DTGGFKPI GHHQTVU KP UGNGEVGF CTGCU QH EWTTGPV KPVGTGUV: ECTDQJ[FTCVG HGTOGPVCVKQPþ RTQFWEVKQP QH HNCXQWT EQORQPGPVUþ DGGT OCVWTCVKQPþ CPF HNCXQWT UVCDKNKV[. $GUKFGU VJGUG VQRKEUþ YG YKNN DTKGHN[ FGÿ UETKDG GHHQTVU VQ KORTQXG DGGT HKNVTCVKQP D[ KPVTQFWEVKQP QH !ÿINWECPQN[VKE GP\[OGU KPVQ DTGYGT•U [GCUVþ CPF VQ KORTQXG UGFKOGPVCVKQP EJCTCEVGTKUVKEU QH XCTKQWU [GCUV UVTCKPU.
5.4 6CTIGVU CPF CRRTQCEJGU KP DTGGFKPI QH DTGYGT•U [GCUV 5.4.1. (GGFKPI VJG DGCUV: ECTDQJ[FTCVG HGTOGPVCVKQP 9JGP RTQFWEKPI DGGTþ VJG OCKP CKO KU VQ EQPUWOG CU OWEJ ECTDQJ[FTCVG UWDUVTCVG CU RQUUKDNGþ CU qWKEMN[ CU RQUUKDNG. #RRTQZKOCVGN[ ô5ó QH YQTV ECTDQJ[FTCVGU CTG PQPÿHGTOGPVCDNGþ OCKPN[ FGZVTKPU QH XCT[KPI NGPIVJþ YJKNG VJG TGOCKPKPI 75ó CTG
15ô
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
HGTOGPVCDNGþ CPF EQPUKUV QH VJG RQN[ÿ CPF QNKIQUCEEJCTKFGU OCNVQUGþ INWEQUGþ UWÿ ETQUGþ HTWEVQUGþ CPF OCNVQVTKQUG. &WTKPI HGTOGPVCVKQPþ OCNVQUG CPF OCNVQVTKQUG WVKNK\CVKQP FQGU PQV UVCTV WPVKN CRRTQZ. 5ýó QH VJG YQTV INWEQUG JCU DGGP WUGF üSVGYCTV GV CN. 1û83ú. 6JKU KU FWG RCTVN[ VQ INWEQUG TGRTGUUKQP QH VJG IGPGU HQT WRÿ VCMG CPF WVKNK\CVKQP QH OCNVQUG CPF OCNVQVTKQUGþ CPF RCTVN[ VQ INWEQUGÿOGFKCVGF KPÿ CEVKXCVKQP QH VJG VTCPURQTVGT RTQVGKPU üHQT CP GZVGPUKXG QXGTXKGY QH VJG IGPGVKEU QH OCNVQUG WVKNK\CVKQP KP DCMGT•U CPF DTGYGT•U [GCUVþ UGG %JCRVGT 3ú. 6JKU RJGPQOGÿ PQP OC[ DG QH RCTVKEWNCT KORQTVCPEG KP JKIJÿITCXKV[ QT UWRGTÿJKIJÿITCXKV[ HGTOGPÿ VCVKQPUþ KP YJKEJ JKIJ COQWPVU QH INWEQUG CTG RTGUGPV. +V JCU DGGP TGRQTVGF VJCV CNG DTGYKPI [GCUV UVTCKPU VJCV YGTG TGUKUVCPV VQ VJG PQPÿOGVCDQNKUCDNG INWEQUG CPCÿ NQIWG ôÿFGQZ[INWEQUG EQWNF OGVCDQNK\G OCNVQUG CPF OCNVQVTKQUG KP VJG RTGUGPEG QH INWEQUG üSVGYCTV GV CN. 1û85ú. +P C OQTG VCTIGVGF CRRTQCEJþ KV YCU HQWPF VJCV QXGTÿ GZRTGUUKQP QH C OCNVQUG VTCPURQTVGT IGPG üMALTú HTQO C IGPG RTQOQVGT PQV TGÿ RTGUUGF D[ INWEQUGþ FWTKPI JKIJ ITCXKV[ HGTOGPVCVKQPþ TGUWNVGF KP C UKIPKHKECPVN[ KPETGCUGF URGGF QH OCNVQUG TGOQXCN HTQO VJG YQTVþ CEEQORCPKGF D[ C EQTTGURQPFÿ KPI FGETGCUG KP VJG VQVCN HGTOGPVCVKQP VKOG ü-QFCOC GV CN. 1ûû5ú. 9JCV CDQWV VJG FGZVTKPU VJCV EQORTKUG VJG ô5ó PQPÿHGTOGPVCDNG YQTV ECTDQJ[ÿ FTCVGU! (KTUVþ VJG FGZVTKPU TGRTGUGPV CP WPWUGF GVJCPQN RTGEWTUQTþ DWV RGTJCRU OQTG KORQTVCPVN[ VJG[ RQUG C RTQDNGO YJGP VJG DTGYGT•U IQCN KU C NQYÿECNQTKG DGGTþ C RTQFWEV V[RG VJCV KU KPETGCUKPIN[ RQRWNCT KP OCP[ RCTVU QH VJG YQTNF. &GZVTKPU CTG DTCPEJGF QNKIQUCEEJCTKFG DTGCMFQYP RTQFWEVU QH VJG OCNV UVCTEJþ EQPVCKPKPI DQVJ "ÿ1þ4ÿ CPF "ÿ1þ÷ÿIN[EQUKFKE NKPMCIGU. )NWEQCO[NCUGU YKVJ "ÿ1þ4 J[FTQN[VKE CEVKXKV[ ECP ECVCN[\G VJG TGNGCUG QH INWEQUG HTQO VJG PQPÿTGFWEKPI GPFU QH VJG RQN[UCEEJCÿ TKFGUþ CPF C UWDUGV QH VJGUG GP\[OGU GXGP RQUUGUU UQOG FGDTCPEJKPI "ÿ1þ÷ J[FTQÿ N[VKE CEVKXKV[. "ÿ#O[NCUGUþ QP VJG QVJGT JCPFþ ECVCN[\G GPFQCO[NQN[VKE ENGCXCIG QH "ÿ1þ4 NKPMCIGU CV C TCVJGT TCPFQO DCUKUþ TGUWNVKPI KP VJG TGNGCUG QH QNKIQUCEEJCÿ TKFGU. 9JGP RTGUGPV VQIGVJGTþ VJG FKHHGTGPV V[RGU QH CO[NQN[VKE GP\[OGU YQWNF qWKEMN[ TGFWEG OQUV QH VJG TGOCKPKPI DGGT FGZVTKPU VQ HGTOGPVCDNG ECTDQJ[FTCVGUþ CPF KPFGGF OQUV NQYÿECNQTKG DGGT VQFC[ KU DGKPI OCFG D[ VJG CFFKVKQP QH ETWFG OKÿ ETQDKCN RTGRCTCVKQPU EQPVCKPKPI UWEJ GP\[OCVKE CEVKXKVKGU. +P CVVGORVU VQ QDVCKP VCTIGVGF UQNWVKQPU VQ VJKU RTQEGUU RTQDNGOþ OWEJ TGUGCTEJ JCU HQEWUGF QP EQPUVTWEVKPI DTGYKPI [GCUVU VJCV ECP UGETGVG CEVKXG HWPICN INWEQCÿ O[NCUGU. 6QIGVJGTþ VJG XCTKQWU GZRGTKOGPVU RGTHQTOGF JCXG UQNXGF OQUV QH VJG RTQDNGOU CUUQEKCVGF YKVJ VJG EQPUVTWEVKQP QH CP CO[NQN[VKE DTGYKPI [GCUV. +PKVKCN UVTCVGIKGU TGNKGF QP TCTG OCVKPI QT RTQVQRNCUV HWUKQP QH FKCUVCVKE YKNF [GCUVU YKVJ DTGYKPI UVTCKPUþ HQNNQYGF D[ DCEMETQUUKPI ü©OGKU 1û71î 6WDD GV CN. 1û81î (TGGÿ OCP 1û81úþ DWV CU QVJGT WPYCPVGF VTCKVU üG.I. VJG RTQFWEVKQP QH ÉRJGPQNKE QHHÿ HNCXQWTÌú YGTG QHVGP CUUQEKCVGF YKVJ VJG YKNF [GCUVUþ VJG GOGTIKPI OQFGTP OQÿ NGEWNCT DTGGFKPI VGEJPKqWGU DGECOG qWKVG C DTGCMVJTQWIJ KP VJKU CTGC QH TGUGCTEJ. )QQF TGUWNVU YKVJ FGZVTKP HGTOGPVCVKQP YGTG ICKPGF WUKPI JKIJÿEQR[ QT NQYÿ EQR[ RNCUOKF GZRTGUUKQP QH XCTKQWU INWEQCO[NCUG IGPGU ü/GCFGP CPF 6WDD 1û85î 2GTT[ CPF /GCFGP 1û88î òCMGTKC CPF *KPEJNKHHG 1û8ûî òCMGTKC GV CN. 1ûû÷î SCMCK GV CN. 1û8ûî 2CTM GV CN. 1ûûýî -KO GV CN. 1ûû4úþ DWV VJG KPVGITCVKXG CRRTQCEJGU ü;QEWO 1û8÷î LCPECUJKTG GV CN. 1û8ûî 2CTM GV CN. 1ûûýî )QRCN CPF *COOQPF 1ûûôú UGGO VQ DG OQTG RTQOKUKPI. 6JWUþ UKIPKHKECPV FGZVTKP HGTOGPVCVKQP CPF 1ýó KPETGCUG KP GVJCPQN RTQFWEVKQP YCU QDVCKPGF CHVGT OWNVKRNG KPVGITCVKQP QH S. cere-
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
Diacetyl
BAP2 etc.
153
2,3-Pentanedione
Threonine Leucine
Valine
Pyruvate
Isoleucine
*-Ketobutyrate ILV2
ILV2
*-Isopropylmalate
*-Acetolactate
Æ-Isopropylmalate
*,Æ-Dihydroxyisovalerate
*-Aceto-*-hydroxybutyrate ILV5
*-Ketoisocaproate
LEU4
ILV5
*,Æ-Dihydroxy-Æ-methylvalerate ILV3
ILV3
*-Ketoisovalerate
*-Keto-Æ-methylvalerate
Isovaleraldehyde
Isobutyraldehyde
*-Methylbutyraldehyde
Isoamyl alcohol
Isobutyl alcohol
ATF1, ATF2
Isoamyl acetate IAH1
Isoamyl alcohol
D-amyl alcohol (2-Methyl butanol)
ATF1, ATF2
Isobutyl acetate IAH1
Isobutyl alcohol
ATF1, ATF2
D-amyl acetate IAH1
D-amyl alcohol
(KI. 2. $KQU[PVJGUKU QH DTCPEJGF EJCKP COKPQ CEKFUþ CPF HQTOCVKQP QH FKCEGV[Nþ HWUGN CNEQÿ JQNUþ CPF GUVGTU D[ Saccharomyces [GCUVU. 0COGU QH VJG EQTTGURQPFKPI IGPGU CTG KPFKECVGF QPN[ KH OGPVKQPGF KP VJG VGZV.
visiae XCT. diastaticus STA1 üDEX2ú KPVQ VJG IGPQOKE T&0# IGPGU ü2CTM GV CN. 1ûûýú. +PUGTVKQP QH VJG Aspergillus niger )# IGPG KP CNN VJTGG IGPQOKE EQRKGU QH VJG DTGYKPI [GCUV HO IGPG TGUWNVGF KP C qWKVG KORTGUUKXG 5ýó FGETGCUG KP FGZVTKP EQPVGPV CPF ôýó KPETGCUG KP GVJCPQN RTQFWEVKQP ü;QEWO 1û8÷î )QRCN CPF *COOQPF 1ûûôú. 6JG A. niger INWEQCO[NCUG KU VJGTOQUVCDNGþ CPF JGPEG PQV KPCEÿ VKXCVGF D[ RCUVGWTK\CVKQPþ DWV VJKU RTQDNGO YCU CXQKFGF KP [GV CPQVJGT GZRGTKOGPV D[ VJG WUCIG QH C VJGTOQÿNCDKNG INWEQCO[NCUG üLCPECUJKTG GV CN. 1û8ûú. 5.4.2 (NCXQWT EQORQPGPVU: VQQ NKVVNG CPF VQQ OWEJ 9JCV CTG VJG EJGOKECN EQORQPGPVU VJCV OCMG WR DGGT HNCXQWT CPF YJGTG FQ VJG[ EQOG HTQO! 9JKNG OCP[ KORQTVCPV EQORQWPFU CTG FGTKXGF HTQO VJG DCTNG[ CPF JQRUþ C HGY ITQWRU QH [GCUV OGVCDQNKVGU VCMG RTQOKPGPV TQNGU KP HQTOCVKQP QH DGGT HNCXQWT. õPG UWEJ ITQWR EQPUKUVU QH VJG JKIJGT CNEQJQNU üCNUQ ECNNGF HWUGN CNEQJQNUú CPF VJGKT CEGVCVG GUVGTUþ YJKEJ CTG KPVGTOGFKCVGU KP VJG OGVCDQNKUO QH VJG DTCPEJGFÿ EJCKP COKPQ CEKFU NGWEKPGþ XCNKPGþ CPF KUQNGWEKPG üUGG (KI. ôú. 6JG EQORQWPF QH JKIJGUV KORCEV KU RTQDCDN[ KUQCO[N CEGVCVGþ QT 3ÿOGVJ[NDWV[N CEGVCVGþ YJKEJ OC[
154
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
KORCTV C FKUVKPEVKXG DCPCPC CPF RGCT HNCXQWT VQ DGGT CV VJG EQPEGPVTCVKQPU QHVGP RTQFWEGF D[ DTGYGT•U [GCUV. +UQCO[N CEGVCVG KU HQTOGF VJTQWIJ GUVGTKHKECVKQP QH 3ÿ OGVJ[N DWVCPÿ1ÿQNþ KUQCO[N CNEQJQNþ C EQORQWPF VJCV OC[ KVUGNH EQPVTKDWVG C UQNÿ XGPVÿNKMG HNCXQWT VQ DGGT. 6JG CNEQJQN KU HQTOGF HTQO KUQXCNGTCNFGJ[FGþ YJKEJ KP VWTP EQOGU HTQO "ÿMGVQKUQECRTQCVGþ VJG FKTGEV RTGEWTUQT HQT NGWEKPG DKQU[PVJGUKU. 6JG KUQCO[N CEGVCVG RTGEWTUQT "ÿMGVQKUQECRTQCVG KU HQTOGF KP VJTGG GP\[OCVKE UVGRU HTQO QH "ÿMGVQKUQXCNGTCVGþ CPF LEU4 GPEQFGU VJG HKTUV QH VJGUG GP\[OGUþ "ÿ KUQRTQR[NOCNCVG U[PVJCUG. 6JKU GP\[OG KU PQTOCNN[ HGGFDCEMÿKPJKDKVGF D[ NGWEKPGþ DWV NGWEKPGÿKPUGPUKVKXG XGTUKQPU QH VJG GP\[OG JCXG DGGP HQWPF KP OWVCPV [GCUVU TGUKUVCPV VQYCTFU VJG VQZKE NGWEKPG CPCNQIWG 5þ5þ5ÿVTKHNWQTQÿ&LÿNGWEKPG ü6(Lú üSCV[CPCTC[CPC GV CN. 1û÷8úþ CPF EQORCTCDNG 6(LÿTGUKUVCPV OWVCPVU QH sake [GCUV JCF KPETGCUGF RTQFWEVKQP QH KUQCO[N CNEQJQN CPF KVU CEGVCVG GUVGT ü#UJKFC GV CN. 1û87ú. +P CEEQTFCPEG YKVJ VJKUþ QXGTGZRTGUUKQP QH LEU4 HTQO OWNVKRNGþ EJTQOQÿ UQOCNN[ KPVGITCVGF EQRKGU KP CP S. cerevisiae sake [GCUV ECWUGF 6(LÿTGUKUVCPEG ü*KTCVC CPF *KTQK 1ûû1ú CPF KPETGCUGF RTQFWEVKQP QH KUQCO[N CNEQJQN ü*KTCVC GV CN. 1ûûôú. 9KVJ VJG UCOG UVTCVGI[þ OWVCPVU QH NCIGT DTGYKPI [GCUV YKVJ JKIJ RTQFWEÿ VKQP QH KUQCO[N CNEQJQN CPF KVU CEGVCVG GUVGT üLGG GV CN. 1ûû5ú YGTG QDVCKPGF. õP VJG QVJGT JCPFþ "ÿMGVQKUQECRTQCVG OC[ CNUQ DG HQTOGF D[ VTCPUCOKPCVKQP QH NGWÿ EKPG VCMGP WR D[ VJG [GCUVþ CPF C RQUKVKXG KPHNWGPEG QH QXGTGZRTGUUKQP QH VJG NGWÿ EKPG VTCPURQTVGT BAP2 QP KUQCO[N CNEQJQN RTQFWEVKQP JCU DGGP UGGP ü-QFCOC GV CN. ôýý1ú. 6YQ CNEQJQN CEGV[NVTCPUHGTCUG IGPGUþ ATF1 CPF ATF2þ CTG RTGUGPV KP Saccharomyces [GCUVU ü(WjKK GV CN. 1ûû4î 0CICUCYC GV CN. 1ûû8úþ CPF VJGKT EQTTGURQPFÿ KPI GP\[OGU CEEQWPV HQT OQUV QH VJG KUQCO[N CEGVCVG HQTOGFþ CU YGNN CU QVJGT CEGÿ VCVG GUVGTU. #P QDXKQWU YC[ VQ CNVGT KUQCO[N CEGVCVG RTQFWEVKQP YQWNF VJWU DG OCPKRWNCVKQP QH VJGUG IGPGU. +PFGGFþ QXGTGZRTGUUKQP QH ATF1 KP S. cerevisiae KPÿ ETGCUGFþ CPF FKUTWRVKQP QH VJG IGPG FGETGCUGFþ RTQFWEVKQP QH KUQCO[N CEGVCVG ü(WjKK GV CN. 1ûû÷ú. (KPCNN[þ KV OC[ DG RQUUKDNG VQ EQPVTQN VJG RTQFWEVKQP QH GUVGTU D[ CNVGTKPI VJG GUÿ VGTCUG CEVKXKV[. 6JWUþ KV YCU HQWPF VJCV KUQCO[N OQPQHNWQTQCEGVCVGÿTGUKUVCPV OWVCPV sake [GCUV JCF CP 85ó TGFWEVKQP KP GUVGTCUG CEVKXKV[ CPF C EQPEQOKVCPV 1.5 VQ ô.ýÿ HQNF KPETGCUG KP HQTOCVKQP QH KUQCO[N CPF KUQDWV[N CEGVCVG ü9CVCPCDG GV CN. 1ûû3úþ CPF VJCV VJG EQR[ PWODGT QH VJG KUQCO[N CEGVCVG GUVGTCUG IGPG IAH1 KP C sake [GCUV KPHNWGPEGF KUQCO[N CEGVCVG RTQFWEVKQP ü(WMWFC GV CN. 1ûû8ú. SWNRJWT EQORQWPFU HQTO CPQVJGT ITQWR QH KORQTVCPV DGGT HNCXQWT EQORQPGPVU. 6JG KORCEV QH GCEJ QH VJGUG EQORQWPFU KU WUWCNN[ EQPUKFGTGF VQ DG PGICVKXGþ DWV OCP[ QH VJGO OC[ YGNN VCMG RCTV KP VJG VQVCN PQTOCN DGGT HNCXQWT. *[FTQIGP UWNÿ RJKFGþ *ôSþ KU C EQORQWPF QH OCjQT KORQTVCPEGþ RTKOCTKN[ FWG VQ VJG WPRNGCUCPV HNCXQWT KV KORCTVUþ DWV CNUQ DGECWUG QH VJG VJKQNU DGNKGXGF VQ DG FGTKXGF HTQO KVþ OGVJCPGVJKQN CPF GVJCPGVJKQN ü6G\WMC GV CN. 1ûûôî 9CNMGT CPF SKORUQP 1ûû3ú üUGG (KI. 3ú. +PUKFG VJG [GCUV EGNNþ *ôS KU EQPFGPUGF VQ CEVKXCVGF JQOQUGTKPGþ C ETWEKCN TGCEVKQP VJCV GPUWTGU CUUKOKNCVKQP QH KPQTICPKE UWNRJWT KPVQ KORQTVCPV QTICPKE UWNÿ RJWT EQORQWPFU UWEJ CU E[UVGKPGþ OGVJKQPKPGþ INWVCVJKQPG CPF Sÿ CFGPQU[NOGVJKQPKPG. /GVCDQNKECNN[ WRUVTGCOþ *ôS KU VJG RTQFWEV QH UWNRJKVG TGFWEÿ VKQPþ CPF KP VWTPþ UWNRJKVG KU FGTKXGF VJTQWIJ VYQ KPVGTOGFKCVGU HTQO UWNRJCVG KQPU
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
155
Sulphate
Thiols
MET3 MET14 Homoserine
Sulphite
MET10
MET2 O-Acetyl-
homoserine
Transcriptional repression of MET genes
MET25 Homocysteine
Hydrogen sulphide Methionine
SAM
SAH
STR4
Cysteine
Glutathione
(KI. 3. SWNRJWT CUUKOKNCVKQP CPF DKQU[PVJGUKU QH UWNRJWTÿEQPVCKPKPI COKPQ CEKFU D[ Saccharomyces [GCUVU. 0COGU QH VJG EQTTGURQPFKPI UVTWEVWTCN IGPGU CTG KPFKECVGF QPN[ KH OGPÿ VKQPGF KP VJG VGZV. S#/þ SÿCFGPQU[N OGVJKQPKPGî S#*þ SÿCFGPQU[N JQOQE[UVGKPG.
VCMGP WR HTQO VJG UWTTQWPFKPIU. # HGY VCTIGVGF CRRTQCEJGU VQ EQPVTQNNKPI *ôS RTQFWEVKQP JCXG DGGP CVVGORVGF. 6JWUþ õOWTC et al. ü1ûû5ú KPETGCUGF VJG EQR[ PWODGT QH MET25þ VJG IGPG GPÿ EQFKPI JQOQE[UVGKPG U[PVJGVCUGþ CPF VJKU TGUWNVGF KP C 1ýÿHQNF TGFWEVKQP KP VJG COQWPV QH J[FTQIGP UWNRJKFG NKDGTCVGF VQ VJG ITQYVJ OGFKWOþ UWRRQUGFN[ FWG VQ GPJCPEGF *ôS EQPFGPUCVKQP. +P C UQOGYJCV UKOKNCT CRRTQCEJþ VJG STR4 (CYS4þ NSH5) IGPGþ GPEQFKPI E[UVCVJKQPKPG !ÿU[PVJCUGþ YCU QXGTGZRTGUUGF KP DTGYGT•U [GCUV ü6G\WMC GV CN. 1ûûôú. 6JKU GP\[OG EQPXGTVU JQOQE[UVGKPG KPVQ E[UVCVJKQPKPGþ CPF KV YCU J[RQVJGUK\GF VJCV QXGTGZRTGUUKQP QH VJG STR4 IGPG YQWNF ECWUG CP KPÿ ETGCUG KP JQOQE[UVGKPG U[PVJGUKU FWG VQ FKXGTUKQP QH JQOQE[UVGKPGþ TGUWNVKPI KP C FTQR KP *ôS CEEWOWNCVKQP. íWKVG CU GZRGEVGFþ *ôS CEEWOWNCVKQP YCU UWRRTGUUGF. 2CTV QH VJG TGCUQP HQT VJKUþ JQYGXGTþ OC[ DG VJCV VJG KPVTCEGNNWNCT EQPEGPVTCVKQP QH E[UVGKPG KU KPETGCUGFþ ECWUKPI GPJCPEGF TGRTGUUKQP QH VJG UWNRJWT CUUKOKNCVKQP IGPGUþ KPENWFKPI VJQUG NGCFKPI VQ *ôS HQTOCVKQP ü2CU\GYUMK CPF õPQ 1ûûôî õPQ GV CN. 1ûû÷î *CPUGP CPF ,QJCPPGUGP ôýýýú. (KPCNN[þ FKUTWRVKQP KP DTGYGT•U [GCUV QH VJG MET10 IGPGUþ GPEQFKPI RCTV QH VJG UWNRJKVG TGFWEVCUG GP\[OG ü*CPUGP GV CN. 1ûû4úþ TGUWNVGF KP C FTCOCVKE KPETGCUG KP UWNRJKVG RTQFWEVKQP CPF C PGCT GNKOKPCÿ VKQP QH UWNRJKFG RTQFWEVKQP ü*CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû5î 1ûû÷Dú.
15÷
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
6JG MG[ EQORQWPF KP VJG UOGNN QH EQQMGF UYGGV EQTPþ &/S üFKOGVJ[NUWNÿ RJKFGúþ KU CPQVJGT GZCORNG. 9JKNG DGKPI ENGCTN[ QDjGEVKQPCDNG CV JKIJ EQPEGPVTCÿ VKQPUþ VJG V[RKECN HNCXQWT QH UQOG NCIGT DTCPFU KU DGNKGXGF VQ DG RCTVN[ FGRGPFGPV QP UOCNN COQWPVU QH &/S. #U VJKU EQORQWPF KU QHVGP HQWPF KP EQPEGPVTCVKQPU jWUV CTQWPF KVU HNCXQWT VJTGUJQNFþ VJG RJ[UKQNQI[ QH KVU RTQFWEVKQP KU QH EQPEGTP. 9JKNG UQOG &/S EQOGU FKTGEVN[ HTQO VJG YQTVþ VJG OCjQTKV[ QH &/S HQWPF KP VJG HKPÿ KUJGF RTQFWEV TGRTGUGPVU C DTGYKPI [GCUV OGVCDQNKVG ü#PPGUU GV CN. 1û7ûî LGGOCPU GV CN. 1ûû3î *CPUGP 1ûûûî *CPUGP GV CN. ôýýôúþ TGUWNVKPI HTQO &/Sõ üFKOGVJ[Nÿ UWNRJQZKFGú TGFWEVKQP D[ VJG OGVJKQPKPG UWNRJQZKFG TGFWEVCUGþ GPEQFGF D[ MXR1 ü*CPUGPþ 1ûûûú. ùUKPI VJG VYQÿUVGR FGNGVKQP RTQEGFWTGþ C NCIGT DTGYKPI [GCUV YKVJ XKTVWCNN[ CDQNKUJGF &/Sõ TGFWEVCUG CEVKXKV[ YCU EQPUVTWEVGF. 6JKU [GCUV RTQFWEGF DGGT YJKEJ JCF C 5ýÿ8ýó NQYGT &/S EQPVGPVþ DWV YCU QVJGTYKUG PQTOCN ü*CPUGP GV CN. ôýýôú. 5.4.3. (NCXQWT UVCDKNKV[: C YC[ VQ KPETGCUGF UJGNH NKHG $GGT KU C UGPUKVKXG DGXGTCIG: KV PGGFU VQ DG MGRV EQQN VQ CXQKF HQTOCVKQP QH CIKPIÿ TGNCVGF QHHÿHNCXQWTU. õH VJGUG VôP ütransÿôÿPQPGPCNúþ FGTKXGF HTQO DCTNG[ HCVV[ CEKF RTGEWTUQTUþ KU VJG OQUV JCTOHWN. 6JG VCUVG VJTGUJQNF QH VôP KU NQYþ KP VJG PCPQITCOöNKVGT TCPIGþ CDQXG YJKEJ VJKU CNFGJ[FG KORCTVU C ECTFDQCTFÿ QT RCRGT[ÿ NKMG EJGOKECN HNCXQWT. # OCjQT VCUM QH VJG DTGYKPI KPFWUVT[ KU VJGTGHQTG VQ MGGR DGGT EQQN CV CNN VKOGUþ UQOGVJKPI VJCV KU qWKVG EQUVN[ CPF QHVGP FKHHKEWNVþ CV NGCUV HQT GZVGPFGF RGTKQFU. *QYGXGTþ PCVWTG JCU RTQXKFGF VJG DTGYKPI [GCUV YKVJ OGCPU VQ FCORGP VJKU RTQDNGOþ CU VôP CPF UWNRJKVG ü*Sõ3ÿú HQTO C HNCXQWTÿKPCEVKXG CFFWEV. #U OGPVKQPGF GCTNKGTþ UWNRJKVG KU C PCVWTCN RTQFWEV QH Saccharomyces [GCUV UWNRJWT CUUKOKNCVKQPþ CPF UQOG KU WUWCNN[ GZETGVGF üUGG CNUQ %JCRVGT 4ú. 6JG COQWPV QH UWNRJKVG RTQFWEGF FGRGPFU QP XCTKQWU HCEVQTU: VJG [GCUV UVTCKP QT KUQNCVG WUGFþ VJG YQTV EQPUVKVWVKQPþ VJG FKOGPUKQPU QH VJG HGTOGPVCVKQP XGUUGNþ VJG HGTOGPVCVKQP TGÿ IKOGþ GVE. ü$TGYGT CPF (GPVQP 1û8ýî 0QTFNøXþ 1û85î &WHQWT GV CN. 1û8ûî )[NNCPI GV CN. 1û8ûúþ CPF CEEQTFKPIN[ VJG COQWPV QH [GCUVÿRTQFWEGF UWNRJKVG KU PQV CNYC[U CFGqWCVG HQT HNCXQWT UVCDKNK\CVKQP. (QT VJKU TGCUQPþ UQOG GHHQTV JCU DGGP RWV KPVQ EQPUVTWEVKQP QH NCIGT DTGYKPI [GCUVU YKVJ KPETGCUGF UWNRJKVG RTQFWEVKQP. 6JG UVTCVGIKGU JCXG DGGP QH VYQ OCKP V[RGU: CDQNKUJOGPV QH VJG CDKNKV[ QH VJG [GCUV VQ VWTP UWNRJKVG KPVQ UWNRJKFGþ CPF KPETGCUG KP VJG OGVCDQNKE HNWZ HTQO UWNÿ RJCVG VQ UWNRJKVG üUGG (KI. 3ú. (QNNQYKPI VJG HKTUV UVTCVGI[þ CNN HQWT IGPQOKE EQRKGU QH MET10þ GPEQFKPI RCTV QH VJG UWNRJKVG TGFWEVCUGþ YGTG KPCEVKXCVGF KP S. carlsbergensis DTGYGT•U [GCUV D[ RCTVKCN FGNGVKQPþ WUKPI VJG VYQÿUVGR IGPG TGRNCEGOGPV RTQEGFWTG. 6JKU TGUWNVGF KP C FTCOCVKE üWR VQ 7ÿHQNFú KPETGCUG KP UWNRJKVG RTQFWEÿ VKQP ü*CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû5î 1ûû÷Dú. # UKFG GHHGEV QH VJKU IGPGVKE CNÿ VGTCVKQP YCU VJCV *ôS HQTOCVKQP YCU CDQNKUJGF. &KUTWRVKQP QH VJG MET2 IGPG KP DTGYGT•U [GCUV YCU CNUQ CEEQORNKUJGFþ CPF NGF VQ NCEM QH OÿCEGV[N JQOQUGTKPGþ C IGPGTCN FGTGRTGUUKQP QH VJG UWNRJCVG CUUKOKNCVKQP CPF VJGTGHQTG JKIJGT UWNRJKVG RTQFWEVKQP ü*CPUGP CPF -KGNNCPFÿ$TCPFV 1ûû5î 1ûû÷Cú. +P VJKU ECUGþ JQYGXGTþ *ôS RTQFWEVKQP YCU CNUQ KPETGCUGF.
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
157
#NQPI VJG NKPGU QH VJG UGEQPF V[RG QH UVTCVGI[þ VJG MET3 IGPG üGPEQFKPI #62 UWNRJWT[NCUGú CPF VJG MET14 IGPG üGPEQFKPI #2S MKPCUGú YGTG QXGTGZRTGUUGF KP S. cerevisiae CPF KP DTGYGT•U [GCUV ü-QTEJ GV CN. 1ûû1ú. 6JG IGPGU YGTG VTCPUETKDGF HTQO VJGKT PCVKXG RTQOQVGTU DWV YKVJ C JKIJN[ KPETGCUGF EQR[ PWODGTþ WUKPI ôµO RNCUOKFU. # EQPUKFGTCDNG KPETGCUG KP UWNRJKVG RTQFWEVKQP YCU QDUGTXGFþ PQV UWTÿ RTKUKPIN[ JKIJGUV KP UVTCKPU FGHKEKGPV KP UWNRJKVG TGFWEVKQP ümet5 OWVCPVUú. +P CEÿ EQTFCPEG YKVJ VJGUG TGUWNVUþ ,QJCPPGUGP ü1ûû4ú HQWPF CP KPETGCUG KP UWNRJKVG RTQÿ FWEVKQP KP C UWNRJKVG TGFWEVCUGÿFGHKEKGPV S. cerevisiae UVTCKPþ CHVGT QXGTGZRTGUUKQP QH CP KPVGITCVGF MET14 IGPG HTQO VJG UVTQPI TPI1 RTQOQVGT. ñGEGPVN[ VJKU UVTCVÿ GI[ YCU GNCDQTCVGF WRQPþ YJGP EQPEQOKVCPV QXGTGZRTGUUKQP QH MET14 CPF VJG IGPG HQT C UWNRJKVG GHHNWZ RWORþ SSU1þ NGF VQ C 1ýÿHQNF KPETGCUG KP UWNRJKVG UGETGÿ VKQP HTQO CP S. cerevisiae UVTCKP ü&QPCNKGU CPF SVCJN ôýýôú. 0QVYKVJUVCPFKPI VJGUG UWEEGUUHWN OGVCDQNKE GPIKPGGTKPI UVTCVGIKGUþ JKIJ RTQÿ FWEVKQP QH UWNRJKVG D[ DTGYGT•U [GCUV OC[ PQV DG DGPGHKEKCN CHVGT CNN. 6JWUþ KV YCU UWIIGUVGF VJCV UWNRJKVG RTQFWEGF FWTKPI HGTOGPVCVKQP EQWNF HQTO PQPÿ OGVCDQNKUCDNG CFFWEVU YKVJ WPYCPVGF ECTDQP[N EQORQWPFU CPF KP VJCV YC[ JKPFGT TGOQXCN QH VJGUG ü&WHQWT GV CN. 1û8ûú. #EEQTFKPI VQ VJKU RQKPV QH XKGYþ UWNRJKVG CFFGF CV VJG RQKPV QH DQVVNKPI OKIJV DG OQTG DGPGHKEKCN VQ VJG DGGT VJCP [GCUVÿ FGTKXGF UWNRJKVG. 6Q VGUV VJKU KFGCþ C OWVCPV DTGYGT•U [GCUV KPCEVKXCVGF KP CNN HQWT EQRKGU QH MET14 YCU EQPUVTWEVGF ü,QJCPPGUGP GV CN. 1ûûûú. 6JG QWVEQOG QH HGTÿ OGPVCVKQP GZRGTKOGPVU YKVJ VJKU [GCUV YCU VJCV UWNRJKVG RTQFWEGF FWTKPI VJG OCKP HGTOGPVCVKQP OKIJV CEVWCNN[ TGUWNV KP C JKIJGT DGGT EQPVGPV QH CEGVCNFGJ[FGþ YJKEJ KU CP QHHÿHNCXQWT CV JKIJ EQPEGPVTCVKQPU. (WTVJGTOQTGþ VJKU GZRGTKOGPV UJQYGF VJCV UWNRJKVG CFFGF CV DQVVNKPI YCU CU GHHKEKGPV CU [GCUVÿFGTKXGF UWNRJKVG KP RTQVGEVKQP CICKPUV VôPþ CPF VJWUþ VQ CXQKF GZEGUUKXG CEGVCNFGJ[FG CEEWOWNCVKQPþ C [GCUV YKVJÿ QWV UWNRJKVG RTQFWEVKQP OC[ UQOGVKOGU EQPUVKVWVG C IQQF UQNWVKQP. 5.4.4 &KCEGV[N CPF OCVWTCVKQP: JQY VQ URGGF VJKPIU WR &KCEGV[N KORCTVU C DWVVGTÿNKMG HNCXQWT VJCV KU WPYCPVGF KP NCIGT DGGTþ CPF KV FGXGNÿ QRU KP DGGT FWG VQ C UNQY PQPÿGP\[OCVKE QZKFCVKXG FGECTDQZ[NCVKQP QH VJG [GCUV OGVCDQNKVG "ÿCEGVQNCEVCVG ü"ÿ#Lú. 6JG OCKP DGPGHKV QH VJG UQÿECNNGF UGEQPFCT[ HGTOGPVCVKQPþ QT OCVWTCVKQP RGTKQFþ KU VQ CNNQY VJKU EQPXGTUKQP VQ VCMG RNCEG CPF VQ CNNQY TGOCKPKPI [GCUV VQ TGOQXG VJG FKCEGV[N. +V ECP DG GEQPQOKECNN[ qWKVG GZRGÿ FKGPV VQ FGETGCUG VJG OCVWTCVKQP RGTKQFî VQ FQ VJCVþ QPG PGGFU VQ FGETGCUG HQTOCÿ VKQP QH "ÿ#L QT URGGF WR KVU TGOQXCN. "ÿ#L KU CP KPVGTOGFKCVG KP VJG XCNKPG DKQU[Pÿ VJGVKE RCVJYC[ üUGG (KI. ôúî ILV2 GPEQFGU CEGVQJ[FTQZ[ CEKF U[PVJCUGþ YJKEJ EQPXGTVU ô OQNGEWNGU QH R[TWXCVG VQ ½ÿ#L CPF ECTDQP FKQZKFG. /QUV "ÿ#L FQGU PQV NGCM QWV QH VJG [GCUV EGNN CPF KU EQPXGTVGF VQ "þ!ÿFKJ[FTQZ[ÿKUQXCNGTCVG D[ VJG CEVKQP QH C TGFWEVQKUQOGTCUG GPEQFGF D[ ILV5. òCNKPG KU U[PVJGUK\GF KP VYQ HWTÿ VJGT GP\[OCVKE UVGRUþ VJG HKTUV QH YJKEJ KU GPEQFGF D[ ILV3. &KCEGV[N VCMGP WR D[ VJG [GCUV EGNN FWTKPI VJG OCVWTCVKQP RGTKQF KU GP\[OCVKECNN[ EQPXGTVGF VQ CEGVQKP CPF HWTVJGT VQ ôþ3ÿDWVCPGFKQN. 6YQ [GCUVÿDCUGF CRRTQCEJGU VQ FGETGCUG VJG OCVWTCÿ VKQP VKOG JCXG DGGP KPXGUVKICVGFþ Kú C FGETGCUG KP VJG TCVG QH "ÿCEGVQNCEVCVG HQTOCÿ VKQP üILV2úþ CPF KKú CP KPETGCUG KP VJG TCVG QH KVU TGOQXCN üILV5ú. # JGVGTQNQIQWU
158
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
CRRTQCEJþ CNUQ FGUETKDGF DGNQYþ KU VJG KPVTQFWEVKQP QH C IGPG HQT C DCEVGTKCN "ÿ CEGVQNCEVCVG FGECTDQZ[NCUG ü"ÿ#L&%ú HQT VJG FKTGEV EQPXGTUKQP QH "ÿ#L KPVQ CEGÿ VQKP. SQOG QH VJG UVTCVGIKGU GZRNQKVGF JCXG DGGP qWKVG UWEEGUUHWN. õPG GCTN[ CRRTQCEJ YCU VJG EQORNGVG TGOQXCN QH CEGVQJ[FTQZ[ CEKF U[PVJCUG CEVKXKV[ KP DTGYGTU• [GCUVþ D[ VJG WUG QH in vitro EQPUVTWEVGF ilv2 FGNGVKQPU ü)jGTÿ OCPUGP GV CN. 1û88ú. 6JKU UVTCVGI[ VWTPGF QWVþ JQYGXGTþ PQV VQ DG RTCEVKECNþ CU XCNKPG WRVCMG KP VJG FGNGVCPVU QH S. carlsbergensis VWTPGF QWV VQ DG KPUWHHKEKGPV ü%. )jGTOCPUGPþ WPRWDNKUJGFþ UGG -KGNNCPFÿ$TCPFV GV CN. 1ûûýú. # NGUU TCFKECN CRÿ RTQCEJ OCFG WUG QH VJG CHQTGOGPVKQPGF KPJKDKVQT UWNHQOGVWTQP OGVJ[N üS/ú üUGG UGEVKQP 5.3ú. S/ÿTGUKUVCPV [GCUV OWVCPVU KPENWFG FQOKPCPV CNNGNGU QH ILV2 ü(CNEQ CPF &WOCU 1û85úþ CPF DCUGF QP VJKU QDUGTXCVKQPþ %. )jGTOCPUGP üWPRWDNKUJGFþ UGG -KGNNCPFÿ$TCPFV GV CN. 1û8ûú FGXKUGF C UVTCVGI[ VQ QDVCKP RCTVKCNN[ DNQEMGF ilv2 OWVCPVU KP NCIGT DTGYKPI [GCUV. 6JKU CRRTQCEJ TGNKGF QP VJG KUQNCVKQP QH URQPVCPGÿ QWU S/ÿTGUKUVCPV OWVCPVU QH CNNQFKRNQKF OGKQVKE NCIGT [GCUV RTQIGP[þ HQNNQYGF D[ ùò OWVCIGPGUKUþ KUQNCVKQP QH S/ÿUGPUKVKXG RUGWFQÿTGXGTVCPVU üECPFKFCVGU HQT JCXÿ KPI QPN[ QPG TGOCKPKPI HWPEVKQPCN ILV2 IGPG NGHVúþ TGRGVKVKQP QH OWVCIGPGUKUþ CPF KUQNCVKQP QH EQNQPKGU YKVJ RCTVKCN QT HWNN TGqWKTGOGPV HQT KUQNGWEKPG CPF XCNKPG. ñGÿ UWNVKPI OWVCPVU YGTG OCVGF VQ TGEQPUVKVWVG CNNQVGVTCRNQKF DTGYKPI [GCUVþ CPF EJCTÿ CEVGTKUVKEU QH UWEJ UVTCKPU KPFKECVG VJCV WUGHWN RTQFWEVKQP UVTCKPU OC[ DG QDVCKPGF D[ VJKU UVTCVGI[. 6JG UGEQPF [GCUVÿDCUGF UVTCVGI[ TGNKGU QP KPETGCUGF EQPXGTUKQP QH "ÿ#L KPVQ "þ!ÿFKJ[FTQZ[ KUQXCNGTCVG D[ KPETGCUKPI VJG COQWPV QH VJG TGURQPUKDNG GP\[OGþ GPEQFGF D[ ILV5 ü)jGTOCPUGP GV CN. 1û88ú. òCTKQWU CRRTQCEJGU JCXG TGNKGF QP JKIJÿEQR[ RNCUOKF GZRTGUUKQP QT IGPQOKE KPVGITCVKQP QH PCVKXG ILV5 IGPGU KP DTGYKPI [GCUVþ D[ YJKEJ WR VQ ÷ýó TGFWEVKQP KP FKCEGV[N RTQFWEVKQP JCU DGGP CEÿ EQORNKUJGF üòKNNCPWGDC GV CN. 1ûûýî )QQUGPU GV CN. 1ûû3î /KVJKGWZ CPF 9GKUU 1ûû5ú. õXGTGZRTGUUKQP QH ILV5 YCU TGRQTVGF VQ TGFWEG FKCEGV[N UWHHKEKGPVN[ VQ OCMG OCVWTCVKQP QH VJG DGGT UWRGTHNWQWU. +PVTQFWEVKQP QH DCEVGTKCN "ÿ#L&% IGPGU KPVQ DTGYKPI [GCUV KU VJG UVTCVGI[ VJCV JCU DGGP IKXGP VJG OQUV CVVGPVKQP. +PKVKCNN[þ GZRTGUUKQP HTQO ôµOÿDCUGF RNCUOKFU YCU VJG OGVJQF QH EJQKEGþ CPF WUKPI [GCUV ADH1 RTQOQVGTÿFKTGEVGF VTCPUETKRVKQP QH VJG Enterobacter aerogenes "ÿ#L&% IGPGþ CP 8÷ó FGETGCUG KP FKCEGV[N RTQFWEVKQP YCU QDVCKPGF üSQPG GV CN. 1û87î SJKOK\W GV CN. 1û8ûú. +P GZVGPUKQPU QH VJGUG GZRGTKOGPVUþ E. aerogenes QT Klebsiella terrigena "ÿ#L&% GZRTGUUKQP YCU FKTGEVGF HTQO GKVJGT VJG ADH1 QT VJG PGK1 RTQOQVGT üSWKJMQ GV CN. 1ûûýúþ CPF KP RKNQV UECNG VGUVKPI QH UQOG QH VJG EQPUVTWEVGF [GCUV UVTCKPU VJG PGGF HQT NCIGTKPI YCU GNKOKPCVGF. 6Q CXQKF RTQDNGOU YKVJ RNCUOKF UVCDKNKV[þ VJG UCOG V[RGU QH EQPÿ UVTWEVU YGTG UVCDN[ KPVGITCVGF KPVQ VJG PGK1 QT ADH1 NQEK KP RQN[RNQKF DTGYGT•U [GCUVþ TGUWNVKPI KP C EQORCTCDNG TGFWEVKQP KP FKCEGV[N RTQFWEVKQP ü$NQOqXKUV GV CN. 1ûû1ú. +P CP CVVGORV VQ OKOKE JKIJÿEQR[ RNCUOKF GZRTGUUKQPþ VJG E. aerogenes "ÿ #L&% IGPG YCU KPVGITCVGF KP OQTG VJCP ôý EQRKGU KP VJG DTGYGT•U [GCUV IGPQOGþ VCMKPI CFXCPVCIG QH VJG JKIJÿEQR[ PWODGT CV VJG RDN1 NQEWU ü(WjKK GV CN. 1ûûýú. 6JG UVTCVGI[ YCU FGXKUGF KP UWEJ C YC[ VJCV WPPGEGUUCT[ DCEVGTKCN &0# UGqWGPEGU YGTG NQUV VJTQWIJ UGEQPFCT[ TGEQODKPCVKQP GXGPVU. 6JG "ÿ#L&% CEVKXKV[ YCU UGGP VQ EQTTGNCVG VQ IGPG EQR[ PWODGTþ CPF RKNQV UECNG VGUVU UJQYGF C UKIPKHKECPV FGÿ ETGCUG KP FKCEGV[N RTQFWEVKQP YKVJ UQOG QH VJG UVTCKPU. 6Q CFFTGUU HWVWTG CRRTQXCN
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
15û
KUUWGUþ VJG "ÿ#L&% IGPG HTQO VJG HQQFÿITCFG QTICPKUO Acetobacter aceti UUR. xylinum JCU CNUQ DGGP VGUVGF ü;COCPQ GV CN. 1ûû4Cþ Dî 1ûû5î 6CFC GV CN. 1ûû5ú. $TGYÿ KPI [GCUV UVTCKPU YKVJ QXGTGZRTGUUKQP QH VJKU IGPG UJQYGF C TGFWEVKQP KP FKCEGV[N RTQFWEVKQP D[ OQTG VJCP 7ýóþ YKVJ VJG TGUWNV VJCV VJG DGGT EQWNF DG RTQFWEGF CV C UKIPKHKECPVN[ UJQTVGPGF VKOG. 5.4.5 SGFKOGPVCVKQP CPF HKNVTCVKQP 0QV QPN[ KU KV KORQTVCPV VJCV VJG [GCUV YKVJKP TGCUQPCDNG VKOG ECP EQPXGTV VJG YQTV EQPUVKVWGPVU KPVQ C DCNCPEGF OKZVWTG QH EQORQWPFU YKVJ VJG FGUKTGF VCUVGþ CTQOC CPF UJGNH NKHGþ DWV VJG [GCUV OWUV CNUQ OGGV FGOCPFU TGNCVGF VQ HNWKF OGEJCPKE CUÿ RGEVU QH OCMKPI DGGT. &WTKPI VJG HKTUV RCTV QH VJG OCKP HGTOGPVCVKQPþ EQPXGEVKQP KP NCTIG DGGT HGTOGPVCVKQP VCPMU KU XKIQTQWUþ CPF XKTVWCNN[ CNN [GCUV KU KP UWURGPUKQP. #V VJG GPF QH VJG OCKP HGTOGPVCVKQPþ QP VJG QVJGT JCPFþ C UWHHKEKGPV COQWPV QH [GCUV OWUV JCXG UGVVNGF VQ DG JCTXGUVGF CPF RKVEJGF KPVQ HTGUJ YQTV VQ UVCTV WR HGTÿ OGPVCVKQP KP QPG QT OQTG PGY VCPMU. *QYGXGTþ WPNGUU CEGVQNCEVCVG CPF FKCEGV[N JCXG CNTGCF[ DGGP UWHHKEKGPVN[ EQPXGTVGF KPVQ CEGVQKP CPF DWVCPGFKQN CV VJG VKOG QH [GCUV JCTXGUVþ C UWKVCDNG PWODGT QH [GCUV EGNNU OWUV TGOCKP KP UWURGPUKQP VQ OGFKÿ CVG VJKU EQPXGTUKQP FWTKPI C UWDUGqWGPV NCIGTKPI RGTKQF. &GXKCVKQPU HTQO VJKU RCVÿ VGTP ECWUG RTQDNGOU. 2TGOCVWTG UGFKOGPVCVKQP OC[ NGCF VQ UYGGV PQV HWNN[ HGTÿ OGPVGF DGGT CPFöQT GZEGUUKXG COQWPVU QH FKCEGV[N. õP VJG QVJGT JCPFþ EGPVTKHWIGU CTG TGqWKTGF VQ JCTXGUV UWHHKEKGPV [GCUV VQ EQPVKPWG TWPPKPI VJG HGTOGPVCVKQP RNCPV KH PQV GPQWIJ [GCUV UGFKOGPVU. +P CFFKVKQPþ NCTIG COQWPVU QH UWURGPFGF [GCUV EGNNU KP VJG OCVWTGF DGGT ECP EQPUVKVWVG C JKIJ CPF EQUVN[ NQCF QP VJG HKNVTCVKQP WPKV. SGFKOGPVCVKQP QH [GCUV KU JKIJN[ FGRGPFGPV QP VJG CDKNKV[ QH VJG EGNNU VQ CFJGTG VQ QPG CPQVJGTþ VQ HNQEEWNCVG. #EEQTFKPIN[þ HNQEEWNCVKQP UJQWNF DG UWHHKEKGPVþ DWV PQV GZEGUUKXGþ CPF UJQWNF JCXG RTQRGT VKOKPI. 6JG OQUV KORQTVCPV OGEJCPKUO HQT HNQEEWNCVKQP QH DTGYGT•U [GCUV KU VJG ECNEKWOÿFGRGPFGPV KPVGTCEVKQP DGVYGGP VJG NGEVKP FQOCKP QH C HNQEEWNKP OQNGEWNG QH QPG EGNN CPF ECTDQJ[FTCVGþ RTGUWOCDN[ QH C PGKIJDQWTKPI EGNN. (NQEEWNGPEG QH KPFWUVTKCN [GCUVU KU QPN[ DTKGHN[ FGUETKDGF JGTGþ DWV KV KU EQXGTGF KP UGXGTCN TGEGPV TGXKGYU üSVTCVHQTF 1ûûôî 6GWPKUUGP CPF SVGGPÿ UOC 1ûû5î &GqWKP ôýý1î 2QNCKPC ôýýô CPF %JCRVGT 4ú. 6JG OQUV GZVGPUKXGN[ FGÿ UETKDGF [GCUV HNQEEWNKP JCU DGGP UJQYP VQ DG NQECVGF CV VJG EGNN UWTHCEG ü$KFCTF GV CN. 1ûû5ú CPF KU GPEQFGF D[ VJG S. cerevisiae FLO1 IGPG üSVGYCTV GV CN. 1û7÷î 6Gÿ WPKUUGP GV CN. 1ûû3î 9CVCTK GV CN. 1ûû4ú. -QDC[CUJK GV CN. ü1ûû8ú OCFG WUG QH EJKÿ OGTCU YKVJ C JQOQNQIQWU IGPG HTQO NCIGT DTGYKPI [GCUV üLg-FLO1ú VQ FGVGTOKPG UKVGU KP VJG NGEVKP FQOCKP KPXQNXGF KP UWICT TGEQIPKVKQPþ UKPEG VJG JQOQNQIQWU HNQEEWNKPU FKHHGT KP VJG RCVVGTP QH KPJKDKVKQP QH HNQEEWNGPEG D[ OQPQJGZQUGU. 6JG 3ÿ& UVTWEVWTG QH VJG NGEVKP FQOCKP QH VJG HNQEEWNKP GPEQFGF D[ Lg-FLO1 KU DGKPI FGVGTOKPGF ü)TQGU GV CN. ôýýôú. #P KORQTVCPV EJCNNGPIG KP VJG KORTQXGOGPV QH HNQEEWNCVKQP EJCTCEVGTKUVKEU QH KPFWUVTKCN [GCUVU KU VQ CEJKGXG CP GZRGFKGPV VKOKPI QH HNQEEWNGPEG. 6Q VJKU GPFþ òGTUVTGRGP GV CN. üôýýýú RNCEGF VJG FLO1 IGPG WPFGT VJG EQPVTQN QH C RTQOQVGT VWTPGF QP KP VJG UVCVKQPCT[ RJCUGî VJG[ KPFGGF CEJKGXGF CP QPUGV QH HNQEEWNCVKQP NCVG KP VJG HGTOGPVCVKQP RTQHKNG.
1÷ý
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
#PQVJGT KORQTVCPV CURGEV QH HNWKF OGEJCPKEU KU XKUEQUKV[ QH YQTV CPF DGGTþ YJKEJ ECP DG GZEGUUKXG KH ¾ÿINWECPU HTQO KPUWHHKEKGPVN[ OQFKHKGF OCNV CTG RTGUGPV. 6JKU ECP ECWUG RTQDNGOU DQVJ KP VJG DTGY JQWUG CPF KP HKNVTCVKQP QH VJG DGGT. +P VJG NCVVGT ECUGþ C DTGYGT•U [GCUV VJCV UGETGVGU C RNCPV ü,CEMUQP GV CN. 1û8÷úþ HWPICN ü2GPVVKNk GV CN. 1û87CþDú QT DCEVGTKCN ü/GNFICCTF GV CN. 1ûû5ú ¾ÿINWECPCUG OC[ TGOÿ GF[ VJG RTQDNGO.
5.5 $TGYGT•U [GCUV KP VJG RQUV-IGPQOKE GTC ©XKFGPVN[þ HWPEVKQPCN IGPGVKEU YKVJ CP[ QTICPKUO IGVU C VTGOGPFQWU DQQUV YJGP VJG EQORNGVG IGPQOG UGqWGPEG QH VJCV QTICPKUO DGEQOGU CXCKNCDNG. Saccharomyces cerevisiae YCU VJG HKTUV GWMCT[QVG VQ DG EQORNGVGN[ UGqWGPEGF ü)QHHGCW GV CN. 1ûû÷úþ CPF VJG CXCKNCDKNKV[ QH VJG FCVC KP RWDNKE FCVCDCUGU qWKEMN[ DGECOG CP KPÿ XCNWCDNG VQQN HQT CP[QPG FQKPI OQNGEWNCT DTGGFKPI QH Saccharomyces URGEKGU. #P[ S. cerevisiaeÿNKMG IGPG EQWNF PQY DG KPCEVKXCVGF D[ FGNGVKQP QT FKUTWRVKQPþ RTQDGU EQWNF DG FGUKIPGF HQT VJG FGVGTOKPCVKQP QH VJG EQPVGPV QH CP[ IKXGP IGPGþ CPFþ KORQTVCPVN[þ VJG UGqWGPEG CNNQYGF TGUGCTEJGTU GZVGPUKXG RTQDKPI QH CURGEVU QH NKHG KP Saccharomyces [GCUVU D[ INQDCN CPCN[UGU QH IGPG CEVKXKV[þ K.G. ÉVJG VTCPÿ UETKRVQOGÌ üUGG CNUQ %JCRVGT 11ú. 9JKNG OWNVKVWFGU QH VTCPUETKRVQOG UVWFKGU JCXG DGGP RGTHQTOGF YKVJ S. cerevisiaeþ VJG UCOG MKPF QH CPCN[UGU YKVJ DTGYKPI [GCUV JCXG DGGP UVCNNGF D[ VJG HCEV VJCV QPN[ TQWIJN[ JCNH QH VJG IGPQOG UGqWGPEG QH VJKU [GCUV KU MPQYP. *QYGXGTþ C HGY UVWFKGU QH VJG S. cerevisiaeÿNKMG ÉJCNHÌ QH VJG S. carlsbergensis VTCPUETKRVQOG JCXG DGGP RGTHQTOGF ü2CPQWVUQRQWNQW GV CN. ôýý1î õNGUGP GV CN. ôýýôî 2WIJ GV CN. ôýýôþ ,COGU GV CN. ôýý3ú. $QVJ UVWFKGU FGUETKDG VJG CEÿ VWCN IGPG GZRTGUUKQP FWTKPI CEVWCN RTQFWEVKQPÿUECNG NCIGT DGGT HGTOGPVCVKQPUþ CPF OC[ RTQXKFG XCNWCDNG KPUKIJV KP VJG KPVTCEGNNWNCT RJ[UKQNQI[ FWTKPI VJG HGTOGPVCVKQP RTQEGUU. SKIPKHKECPVN[þ VJG QDVCKPGF FCVC OC[ CNUQ UWRRN[ VJG OQNGEWNCT DTGGFGT QH DTGYGT•U [GCUV YKVJ C NCTIG COQWPV QH IGPG RTQOQVGTU HQT VJG WUG KP ÉVCKNQTGFÌ GZÿ RTGUUKQP QH IGPGU KP DTGYKPI [GCUVU. õPG OWUV PQV HQTIGVþ JQYGXGTþ VJG J[DTKF PCVWTG QH S. carlsbergensisî K.G. VJGTG KU C YJQNG QVJGT ÉJCNHÌ IGPQOG KP VJGTGþ YJKEJ OC[ DGJCXG qWKVG FKHHGTGPVN[ KP VGTOU QH RCVVGTPU QH IGPG VTCPUETKRVKQP. 6JWUþ VJG UGÿ qWGPEG QH VJG PQPÿS. cerevisiaeÿNKMG RCTV QH VJG DTGYGT•U [GCUV IGPQOG KU PGEGUUCT[ VQ HWNN[ CRRTGEKCVG VJG DGJCXKQWT QH VJKU [GCUV KP KPFWUVTKCN DGGT RTQFWEVKQPþ CPF VQ HWNN[ GZRNQKV VJG RQUUKDKNKVKGU QH ETGCVKPI C ÉEWUVQOK\GFÌ DTGYKPI [GCUV. ©ZEKVKPIN[þ FCVC QH VJKU MKPF OC[ UQQP DG CXCKNCDNGþ CU C EQORNGVG UGqWGPEKPI QH C NCIGT DTGYKPI [GCUV JCU PQY DGGP RGTHQTOGF ü0CMCQ GV CN. &TCHV IGPQOG UGqWGPEG QH VJG NCIGT DTGYKPI [GCUVþ OCPWUETKRV KP RTGRCTCVKQPú.
5.ÿ %QPENWFKPI TGOCTMU +P VJKU EJCRVGTþ YG JCXG FGUETKDGF HWPEVKQPCN IGPGVKEU QH DTGYGT•U [GCUVþ GORJCÿ UK\KPI NCIGT [GCUVþ RJGPQV[RGU TGNCVGF VQ RGTHQTOCPEG DCUGF QP OGVCDQNKUO KP
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
1÷1
RTCEVKECN HGTOGPVCVKQPþ CPF QRVKQPU VJCV MPQYNGFIG KP VJG HKGNF JCXG IKXGP VQ IGÿ PGVKE KORTQXGOGPV. 9G JCXG FGUETKDGF JQY OCP[ RTCEVKECN CURGEVU QH DGGT HGTOGPVCVKQP ECP DG CRÿ RTQCEJGF D[ UVTCKP KORTQXGOGPVþ KPENWFKPI ECTDQJ[FTCVG WVKNK\CVKQPþ HNCXQWT EQPÿ VTQNþ HNCXQWT UVCDKNKV[þ CPF OCVWTCVKQP VKOG. 9G JCXG WPFGTNKPGF VJG KORQTVCPEG QH WPFGTUVCPFKPI VJG IGPGVKE UVTWEVWTG QH VJG QTICPKUO VJCV KU VQ DG KORTQXGFþ OQTG RCTVKEWNCTN[ VJG J[DTKF PCVWTG CPF VJG PWODGT QH EJTQOQUQOG UGVU KP NCIGT DTGYÿ KPI [GCUV. /QUV QH VJG CRRTQCEJGU VQ IGPGVKE KORTQXGOGPV KPXQNXG IGPG VGEJPQNQI[. 2TQUÿ RGEVU QH YJGP FKTGEV WUG QH IGPG VGEJPQNQI[ KP HQQF CPF DGXGTCIG RTQFWEVKQP YKNN DGEQOG IGPGTCNN[ CEEGRVGF D[ VJG OCjQTKV[ QH EQPUWOGTU KU C qWGUVKQP QH VJG JCPÿ FNKPI QH OKUEQPEGRVKQPUþ KPENWFKPI KOCIKPGF UCHGV[ RTQDNGOUþ CPF PQV JCPFNKPI QH TGCN UCHGV[ RTQDNGOU. 0GXGTVJGNGUUþ JQY VJG KUUWG KU KPVTQFWEGF KU QH KORQTVCPEG. +H C DGGT KU OCTMGVGF YJKEJ KU RTQFWEGF WUKPI C ÉIGPGVKECNN[ OQFKHKGFÌ [GCUVþ YJKEJ UCXGU RTQFWEVKQP EQUVU YKVJ PQ EJCPIG KP qWCNKV[þ VJG CXGTCIG EQPUWOGT YKNN JCXG NKVVNG OQVKXCVKQP VQ DW[ VJG DGGT. +V OWUV DG DQTPG KP OKPF VJCV C XGT[ NCTIG RCTV QH VJG RTKEG QH VJG DGGT IGPGTCNN[ EQTTGURQPFU VQ EQUVU KP DQVVNKPIþ VTCPURQTVþ FKUVTKDWÿ VKQPþ JCPFNKPI KP VJG UVQTGþ CPF OCTMGVKPI. 6JWUþ GXGP KH C IKXGP OQFKHKECVKQP [KGNFU EQPUKFGTCDNG UCXKPIU KP PGV RTQFWEVKQP EQUVUþ CPF GXGP KH VJG UCXKPIU CTG HWNN[ VTCPUHGTTGF VQ VJG EQPUWOGTþ UJGöJG OKIJV YGNN HKPF VJG UCXKPIU QH NKVVNG UKIÿ PKHKECPEG. 6JG RTQURGEVU HQT CEEGRVCPEG YKNN DG DGVVGT KH VJG OQFKHKECVKQP TGUWNVU KP C ENGCT KORTQXGOGPV KP UQOG qWCNKV[ RCTCOGVGT QT GPXKTQPOGPVCN KORCEV. 6JG NCEM QH WPFGTUVCPFKPI CPF MPQYNGFIG QH DCUKE DKQNQIKECN RTKPEKRNGU KP VJG IGPGTCN RWDNKE EWTTGPVN[ NKOKVU VJG WUG QH IGPG VGEJPQNQI[ FKTGEVN[ KP HQQF CPF DGXGTCIG RTQFWEVKQP KP OQUV RCTVU QH VJG YQTNF. 9G DGNKGXG VJCV VJG QPN[ NQPIÿ VGTO GHHKEKGPV YC[ VQ RTQOQVG C TCVKQPCN CVVKVWFG VQYCTF IGPG VGEJPQNQI[ KP VJG IGPGTCN RWDNKE KU IQQF UEKGPEG GFWECVKQPþ KPENWFKPI GFWECVKQP KP DKQNQI[þ KP RTKÿ OCT[ CPF UGEQPFCT[ UEJQQNU. 6JKU YQWNF QH EQWTUG DGPGHKV OCP[ QVJGT HKGNFU VJCP VJG WUG QH IGPG VGEJPQNQI[. )GPGVKEU KP KVUGNH EQPVCKPU C UVTQPI CURGEV QH DKQNQIKECN HWPEVKQPþ UKPEG KV TGUVU QP RJGPQV[RKE FKHHGTGPEGU DGVYGGP OWVCPVU CPF VJG YKNF V[RG. +P QWT GPVJWUKCUO CDQWV PGY INQDCN VGEJPKqWGU UWEJ CU IGPQOKEUþ VTCPUETKRVQOKEUþ RTQVGQOKEU CPF OGVCDQNQOKEUþ KV KU KORQTVCPV VQ OCKPVCKP VJG ENCUUKECN XKGY QH MGGRKPI HQEWU QP DKQNQIKECN HWPEVKQPþ QH EQWTUGþ CV VJG UCOG VKOG VCMKPI CFXCPVCIG QH VJG QRRQTVWPKÿ VKGU IKXGP D[ INQDCN RGTURGEVKXGU. +P HCEVþ YG HKPF KV EQPEGKXCDNG VJCV KH YGþ UEKGPÿ VKUVU KPENWFGFþ JCF DGGP OQTG HQEWUGF QP HWPEVKQPCNKV[ QH IGPGVKE EJCPIGU VJCP QP VJG VGEJPKqWGU D[ YJKEJ VJG[ CTG CEJKGXGFþ VJGP VJG IGPGTCN RWDNKE YQWNF JCXG JCF CP GCUKGT VKOG WPFGTUVCPFKPI YJCV YG CTG UC[KPI. +V KU C EJCNNGPIG HQT CNN UEKGPVKUVUþ KPENWFKPI VJQUG YQTMKPI QP DGGT DTGYKPIþ VQ EQOOWPKECVG VQ QWT HGNNQY EKVK\GPU KP WPFGTUVCPFCDNG VGTOU YJCV YG FQþ TCVJGT VJCP HCNNKPI KPVQ VJG VGORVCVKQP QH KOÿ RTGUUKPI VJGO YKVJþ CV VKOGUþ CP GZCIIGTCVGF RKEVWTG QH QWT RTQHGUUKQPCN ECRCDKNKÿ VKGU.
1÷ô
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
#EMPQYNGFIGOGPVU 9G VJCPM &T. ;. -QFCOCþ &T. S. ñCKPKGTKþ &T. ;. -CPGMQþ CPF ;. 0CMCQ HQT CNÿ NQYKPI WU VQ TGHGT VQ WPRWDNKUJGF TGUWNVU.
þGHGTGPEGU #DDQVV /Sþ 2WIJ 6# CPF 2TKPING #6 ü1ûû3ú $KQVGEJPQNQIKECN CFXCPEGU KP DTGYKPI. +P:)WORþ $*þ GF:$GGT CPF 9KPG RTQFWEVKQP #PCN[UKUþ %JCTCEVGTK\CVKQP CPF 6GEJPQNQIKECN #FXCPEGU.þ #OGTKECP %JGOKECN SQEKGV[þ 9CUJKPIVQP &%þ ùS# RR 15ýÿ #MCFC 18ýñþ *KTQUCYC +þ -CYCJCVCþ /þ *QUJKFCþ * CPF 0KUJK\CYCþ ; üôýýôú SGVU QH KPVGITCVÿ KPI RNCUOKFU CPF IGPG FKUTWRVKQP ECUUGVVGU EQPVCKPKPI KORTQXGF EQWPVGTÿUGNGEVKQP OCTMGTU FGUKIPGF HQT TGRGCVGF WUG KP DWFFKPI [GCUV. ;GCUV 1û:3û3ÿ4ýô #PFGTUGP 6*þ *QHHOCPP Lþ )TKHQPG ñþ 0KNUUQPÿ6KNNITGP 6 CPF -KGNNCPFÿ$TCPFV /% üôýýýú $TGYKPI [GCUV IGPGVKEU. ©$% /QPQITCRJ ô8þ (CEJXGTNCI *CPU %CTNþ 0·TPDGTI RR 14ýÿ 147 #PPGUU $,þ $COHQTVJ %9 CPF 9CKPYTKIJV 6 ü1û7ûú 6JG OGCUWTGOGPV QH FKOGVJ[N UWNRJQZKFG KP DCTNG[ CPF OCNV CPF KVU TGFWEVKQP VQ FKOGVJ[N UWNRJKFG D[ [GCUV. , +PUV $TGY 85:34÷ÿ34û #UJKFC Sþ +EJKMCYC ©þ SWIKPCOK - CPF +OC[CUW S ü1û87ú +UQNCVKQP CPF CRRNKECVKQP QH OWÿ VCPVU RTQFWEKPI UWHHKEKGPV KUQCO[N CEGVCVGþ C UCMG HNCXQWT EQORQPGPV. #ITKE $KQN %JGO 51:ôý÷1ÿôý÷5 &G $CTTQU LQRGU /þ $GNNQP ,ñþ SJKTNG[ 0, CPF )CPVGT 2( üôýýôú ©XKFGPEG HQT OWNVKRNG KPÿ VGTURGEKHKE J[DTKFK\CVKQP KP Saccharomyces UGPUW UVTKEVQ URGEKGU. (©/S ;GCUV ñGU 1:3ô3ÿ331 $KFCTF (þ $QP[ /þ $NQPFKP $þ &GqWKP S CPF $CTTG 2 ü1ûû5ú 6JG Saccharomyces FLO1 HNQEEWNCVKQP IGPG GPEQFGU HQT C EGNN UWTHCEG RTQVGKP. ;GCUV 11:8ýûÿ8ôô $KNKPUMK %# CPF %CUG[ )2 ü1û8ûú &GXGNQROGPVU KP URQTWNCVKQP CPF DTGGFKPI QH DTGYGT•U [GCUV. ;GCUV 5:4ôûÿ438 $KNKPUMK %#þ ñWUUGNN + CPF SVGYCTV )) ü1û87ú %TQUUÿDTGGFKPI QH Saccharomyces cerevisiae CPF Saccharomyces uvarum ücarlsbergensisú D[ OCVKPI QH OGKQVKE UGITGICPVU:KUQNCVKQP CPF EJCTCEVGTK\CVKQP QH URGEKGU J[DTKFU. 2TQE ô1UV %QPIT ©WT $TGY %QPXþ 1û87þ /Cÿ FTKF RR 4û7ÿ5ý4 $NQOqXKUV -þ SWKJMQ /Lþ -PQYNGU , CPF 2GPVVKNk / ü1ûû1ú %JTQOQUQOCN KPVGITCVKQP CPF GZRTGUUKQP QH VYQ DCEVGTKCN "ÿCEGVQNCEVCVG FGECTDQZ[NCUG IGPGU KP DTGYGT•U [GCUV. #RRN ©PXKT /KETQDKQN 57:ô7û÷ÿô8ý3 $QGMG ,&þ LC%TQWVG ( CPF (KPM )ñ ü1û84ú # RQUKVKXG UGNGEVKQP HQT OWVCPVU NCEMKPI QTQVKFKPGÿ5•ÿRJQURJCVG FGECTDQZ[NCUG CEVKXKV[ KP [GCUV:5ÿHNWQTQÿQTQVKE CEKF TGUKUVCPEG. /QN )GP )GPGV 1û7:345ÿ34÷ $TGYGTþ ,& CPF (GPVQPþ /S ü1û8ýú 6JG HQTOCVKQP QH UWNRJWT FKQZKFG FWTKPI HGTOGPVCVKQP. 2TQE 1÷VJ %QPX +PUV $TGYþ #WUVTCNKC CPF 0GY
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
1÷3
%CUCTGIQNC Sþ 0IW[GP *òþ LCRCVJKVKU )þ -QV[M # CPF )CKNNCTFKP % üôýý1ú #PCN[UKU QH VJG EQPUVKVWVKQP QH VJG DGGT [GCUV IGPQOG D[ 2%ñ UGqWGPEKPI CPF UWDVGNQOGTKE UGqWGPEG J[DTKFK\CVKQP. +PV , S[UV ©XQN /KETQDKQN 51:1÷ý7ÿ1÷18 %CUG[þ )2 ü1û8÷Cú %NQPKPI CPF CPCN[UKU QH VYQ CNNGNGU QH VJG ILV3 IGPG HTQO Saccharomyces carlsbergensis. %CTNUDGTI ñGU %QOOWP 51:3ô7ÿ341 %CUG[þ )2 ü1û8÷Dú /QNGEWNCT CPF IGPGVKE CPCN[UKU QH EJTQOQUQOGU ï KP Saccharomyces carlsbergensis. %CTNUDGTI ñGU %QOOWP 51:343ÿ3÷ô %CUG[ )2þ ïKCQ 9 CPF ñCPM )* ü1û88ú # EQPXGPKGPV FQOKPCPV UGNGEVKQP OCTMGT HQT IGPG VTCPUHGT KP KPFWUVTKCN UVTCKPU QH Saccharomyces [GCUV:SMRI GPEQFGF TGUKUVCPEG VQ VJG JGTDKEKFG UWNHQOGVWTQP OGVJ[N. , +PUV $TGY û4:û3ÿû7 %QTTCP *S ü1û75ú # *KUVQT[ QH $TGYKPI. &CXKF ì %JCTNGUþ 0GYVQP #DDQVVþ ù&GqWKP S üôýý1ú 6JG RQVGPVKCN QH IGPGVKE GPIKPGGTKPI HQT KORTQXKPI DTGYKPIþ YKPGÿOCMKPI CPF DCMKPI [GCUVU. #RRN /KETQDKQN $KQVGEJPQN 5÷:577ÿ588 &QPCNKGU ù©$ CPF SVCJN ù üôýýôú +PETGCUKPI UWNRJKVG HQTOCVKQP KP SCEEJCTQO[EGU EGTGÿ XKUKCG D[ QXGTGZRTGUUKQP QH MET14 CPF SSU1. ;GCUV 1û:475ÿ484 &WHQWT ,ÿ2þ %CTRGPVKGT $þ -WNCMWODC /þ XCP *CGEJV ,ÿL CPF &GXTGWZ # ü1û8ûú #NVGTCVKQP QH Sõô RTQFWEVKQP FWTKPI HGTOGPVCVKQP. 2TQE ôôPF %QPIT ©WT $TGY %QPXþ 1û8ûþ <·TKEJ RR 331ÿ338 ©OGKU %ÿ% ü1û71ú # PGY J[DTKF [GCUV HQT VJG HGTOGPVCVKQP QH YQTV FGZVTKPU. #O SQE $TGY %JGO 2TQE ôû:58ÿ÷ô (CNEQ S% CPF &WOCU -S ü1û85ú )GPGVKE CPCN[UKU QH OWVCPVU QH Saccharomyces cerevisiae TGUKUVCPV VQ VJG JGTDKEKFG UWNHQOGVWTQP OGVJ[N. )GPGVKEU 1ýû:ô1ÿ35 (TGGOCP ñ( ü1û81ú %QPUVTWEVKQP QH DTGYKPI [GCUVU HQT RTQFWEVKQP QH NQY ECTDQJ[FTCVG DGGTU. 2TQE 18VJ %QPIT ©WT $TGY %QPXþ 1û81þ %QRGPJCIGP RR 4û7ÿ5ý4 (WjKK 6þ -QPFQ -þ SJKOK\W (þ SQPG *þ 6CPCMC ,ÿ+ CPF +PQWG 6 ü1ûûýú #RRNKECVKQP QH C TKDQÿ UQOCN &0# KPVGITCVKQP XGEVQT KP VJG EQPUVTWEVKQP QH C DTGYGT•U [GCUV JCXKPI "ÿ CEGVQNCEVCVG FGECTDQZ[NCUG CEVKXKV[. #RRN ©PXKT /KETQDKQN 5÷:ûû7ÿ1ýý3 (WjKK 6þ 0CICUCYC 0þ +YCOCVUW #þ $QICMK 6þ 6COCK ; CPF *COCEJKþ / ü1ûû4ú /QNGEWNCT ENQPKPIþ UGqWGPEG CPCN[UKUþ CPF GZRTGUUKQP QH VJG [GCUV CNEQJQN CEGV[NVTCPUHGTCUG IGPG. #RRN ©PX /KETQDKQN ÷ý:ô78÷ÿô7ûô (WjKK 6þ ;QUJKOQVQ * CPF 6COCK ; ü1ûû÷ú #EGVCVG GUVGT RTQFWEVKQP D[ SCEEJCTQO[EGU EGTÿ GXKUKCG NCEMKPI VJG ATF1 IGPG GPEQFKPI VJG CNEQJQN CEGV[NVTCPUHGTCUG. , (GTOGPV $KQÿ GPI 81:538ÿ54ô (WMWFC -þ ;COCOQVQ 0þ -K[QMCYC ;þ ;CPCIKWEJK 6þ 9CMCK ;þ -KVCOQVQ -þ +PQWG ; CPF -KOWTC # ü1ûû8ú $CNCPEG QH CEVKXKVKGU QH CNEQJQN CEGV[NVTCPUHGTCUG CPF GUVGTCUG KP Saccharomyces cerevisiae KU KORQTVCPV HQT RTQFWEVKQP QH KUQCO[N CEGVCVG. #RRN ©PXKTQP /KETQDKQN ÷4:4ý7÷ÿ4ý78 )jGTOCPUGP % ü1û83ú /WVCIGPGUKU CPF IGPGVKE VTCPUHQTOCVKQP QH OGKQVKE UGITGICPVU QH NCÿ IGT [GCUV. %CTNUDGTI ñGU %QOOWP 48:557ÿ5÷5 )jGTOCPUGPþ % ü1ûû1ú:%QORCTKUQP QH IGPGU KP Saccharomyces cerevisiae CPF Saccharomyces carlsbergensis. 2J& VJGUKUþ ùPKXGTUKV[ QH %QRGPJCIGPþ 1ûû1þ %QRGPJCIGP )jGTOCPUGP % CPF SKIUICCTF 2 ü1û81ú %QPUVTWEVKQP QH C J[DTKF DTGYKPI UVTCKP QH Saccharomyces carlsbergensis D[ OCVKPI QH OGKQVKE UGITGICPVU. %CTNUDGTI ñGU %QOOWP 4÷:1ÿ11 )jGTOCPUGP %þ 0KNUUQPÿ6KNNITGP 6þ 2GVGTUGP ,)Lþ -KGNNCPFÿ$TCPFV /%þ SKIUICCTF 2 CPF *QNODGTI S ü1û88ú 6QYCTFU FKCEGV[NÿNGUU DTGYGTU [GCUV +PHNWGPEG QH ilv2 CPF ilv5 OWÿ VCVKQPU. , $CUKE /KETQDKQN ô8:175ÿ183 )QHHGCW et al ü1ûû÷ú LKHG YKVJ ÷ýýý IGPGU. SEKGPEG ô74:5÷ôÿ5÷7
1÷4
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
)QNFUVGKP #L CPF /E%WUMGT ,* ü1ûûûú 6JTGG PGY FQOKPCPV FTWI TGUKUVCPEG ECUUGVVGU HQT IGPG FKUTWRVKQP KP Saccharomyces cerevisiae. ;GCUV 15:1541ÿ1553 )QQUUGPU ©þ &GDQWTI #þ òKNNCPWGDC -& CPF /CUUEJGNGKP %# ü1ûû3ú &GETGCUGF FKCEGV[N RTQFWEVKQP KP NCIGT DTGYKPI [GCUV D[ KPVGITCVKQP QH VJG ILV5 IGPG. 2TQE ô4VJ %QPIT ©WT $TGY %QPXþ 1ûû3þ õUNQ RR ô51ÿô58 )QRCN %ò CPF *COOQPF ,ñ/ ü1ûûôú ùUG QH IGPGVKECNN[ÿOQFKHKGF [GCUVU HQT DGGT RTQFWEÿ VKQP. 2TQE 5VJ +PVN $TGY 6GEJ %QPHþ 1ûûôþ *CTTQICVG RR ôû7ÿ3ý÷ )TQGU /þ 6GKNWO -þ õNGUGP -þ 2QWNUGP (/ CPF *GPTKMUGP # üôýýôú 2WTKHKECVKQPþ ET[UVCNKÿ \CVKQP CPF RTGNKOKPCT[ ïÿTC[ FKHHTCEVKQP CPCN[UKU QH VJG ECTDQJ[FTCVGÿDKPFKPI FQOCKP QH HNQEEWNKPþ C EGNNÿCFJGUKQP OQNGEWNG HTQO Saccharomyces carlsbergensis. #EVC %T[UV &58:ô135ÿô137 )[NNCPIþ *þ 9KPIGþ / CPF -QTEJþ % ü1û8ûú ñGIWNCVKQP QH Sõô HQTOCVKQP FWTKPI HGTOGPVCÿ VKQP. 2TQE ôôPF %QPIT ©WT $TGY %QPXþ 1û8ûþ <·TKEJ RR 347ÿ354 *COOQPF ,ñ/ ü1ûû5ú )GPGVKECNN[ÿOQFKHKGF DTGYKPI [GCUVU HQT VJG ô1UV EGPVWT[. 2TQITGUU VQ FCVG. ;GCUV 11:1÷13ÿ1÷ô7 *CPUGP ©% ü1883ú ñGEJGTEJGU UWT NC RJ[UKQNQIKG GV NC OQTRJQNQIKG FGU HGTOGPVU CNEQQNKqWGU ò. /oVJQFGU RQWT QDVGPKT FGU EWNVWTGU RWTGU FG SCEEJCTQO[EGU GV FG OKMTQQTICPKUOGU CPCNQIWGU. %QORV ñGPF 6TCX LCD %CTNUDGTI ô:ûôÿ1ý5 *CPUGP ©% ü1ûý8ú ñGEJGTEJGU UWT NC RJ[UKQNQIKG GV NC OQTRJQNQIKG FGU HGTOGPVU CNEQQNKqWGU ï+++. 0QWXGNNGU oVWFGU UWT FGU NGXWTGU FG DTCUUGTKG i HGTOGPVCVKQP DCUUG. %QORV ñGPF 6TCX LCD %CTNUDGTI 7:17ûÿô17 *CPUGP , ü1ûûûú +PCEVKXCVKQP QH MXR1 CDQNKUJGU HQTOCVKQP QH FKOGVJ[N UWNRJKFG HTQO FKÿ OGVJ[N UWNHQZKFG KP Saccharomyces cerevisiae. #RRN ©PX /KETQDKQN ÷5:3û15ÿ3û1û *CPUGP , CPF ,QJCPPGUGP 2( üôýýýú %[UVGKPG KU GUUGPVKCN HQT VTCPUETKRVKQPCN TGIWNCVKQP QH VJG UWNRJWT CUUKOKNCVKQP IGPGU KP S. cerevisiae. /QN )GP )GPGV ô÷3:535ÿ54ô *CPUGP , CPF -KGNNCPFÿ$TCPFV / ü1ûû4ú Saccharomyces carlsbergensis EQPVCKPU VYQ HWPEÿ VKQPCN MET2 CNNGNGU UKOKNCT VQ JQOQNQIWGU HTQO S cerevisiae CPF S monacensis. )GPG 14ý:33ÿ4ý *CPUGP , CPF -KGNNCPFÿ$TCPFV /% ü1ûû5ú )GPGVKE EQPVTQN QH UWNRJKVG RTQFWEVKQP KP DTGYGT•U [GCUV. 2TQE ô5VJ %QPIT ©WT $TGY %QPX 1ûû5þ $TWUUGNU RR 31ûÿ3ô8 *CPUGP , CPF -KGNNCPFÿ$TCPFV /% ü1ûû÷Cú +PCEVKXCVKQP QH MET2 KP DTGYGT•U [GCUV KPÿ ETGCUGU VJG NGXGN QH UWNRJKVG KP DGGT. , $KQVGEJPQN 5ý:75ÿ87 *CPUGP , CPF -KGNNCPFÿ$TCPFV /% ü1ûû÷Dú +PCEVKXCVKQP QH MET10 KP DTGYGT•U [GCUV URGÿ EKHKECNN[ KPETGCUGU Sõô HQTOCVKQP FWTKPI DGGT RTQFWEVKQP. 0CVWTG $KQVGEJPQN 14:1587ÿ 15û1 *CPUGP , CPF -KGNNCPFÿ$TCPFV /% ü1ûû÷Eú /QFKHKECVKQP QH DKQEJGOKECN RCVJYC[U KP KPFWUÿ VTKCN [GCUVU. , $KQVGEJPQN 4û:1ÿ1ô *CPUGP ,þ %JGTGUV * CPF -KGNNCPFÿ$TCPFV /% ü1ûû4ú 6YQ FKXGTIGPV MET10 IGPGUþ QPG HTQO Saccharomyces cerevisiae CPF QPG HTQO Saccharomyces carlsbergensisþ GPEQFG VJG " UWDWPKV QH UWNRJKVG TGFWEVCUG CPF URGEKH[ RQVGPVKCN DKPFKPI UKVGU HQT (#& CPF 0#&2*. , $CEVGTKQN 17÷:÷ý5ýÿ÷ý58 *CPUGP ,þ $TWWP Sòþ $GEJ L/ CPF )jGTOCPUGP % üôýýôú $TGYKPI [GCUV GZRTGUUKQP QH VJG MXR1 IGPG KU VJG OCjQT FGVGTOKPCPV HQT VJG EQPVGPV QH FKOGVJ[N UWNRJKFG KP DGGT. (©/S ;GCUV ñGU ô:137ÿ14û *CUJKFCÿõMCFQ 6þ õICYC #þ -CVQ + CPF 6CMGUCMQ - ü1ûû8ú 6TCPUHQTOCVKQP U[UVGO HQT RTQVQVTQRJKE KPFWUVTKCN [GCUVU WUKPI VJG AUR1 IGPG CU C FQOKPCPV UGNGEVKQP OCTMGT. (©$S LGVV 4ô5:117ÿ1ôô
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
1÷5
*GPFGTUQP ñ%#þ %QZ $S CPF 6WDD ñ ü1û85ú 6JG VTCPUHQTOCVKQP QH DTGYKPI [GCUVU YKVJ C RNCUOKF EQPVCKPKPI VJG IGPG HQT EQRRGT TGUKUVCPEG. %WTT )GPGV û:133ÿ138 *KPEJNKHHG © ü1ûû1ú SVTCKP KORTQXGOGPV QH DTGYKPI [GCUV. +P:2GDGTF[þ ,(þ %CVGPþ %©þ õIFGPþ ,© CPF $GPPGVVþ ,9þ GFU:#RRNKGF /QNGEWNCT )GPGVKEU QH (WPIK S[ORQUKWO QH VJG $TKVKUJ /[EQNQIKECN SQEKGV[ JGNF CV VJG ùPKXGTUKV[ QH 0QVVKPIJCOþ #RTKN 1ûûýþ $TKVKUJ /[EQNQIKECN SQEKGV[þ %CODTKFIGþ RR 1ôûÿ145 *KTCVC & CPF *KTQK 6 ü1ûû1ú )GPGU VJCV ECWUG QXGTRTQFWEVKQP QH KUQCO[N CNEQJQN D[ KPÿ ETGCUGF IGPGÿFQUCIG KP Saccharomyces cerevisiae. #ITKE $KQN %JGO 55:û1ûÿûô4 *KTCVC &þ #QMK Sþ 9CVCPCDG -þ 6UWMKQMC / CPF SW\WMK 6 ü1ûûôú SVCDNG QXGTRTQFWEVKQP QH KUQCO[N CNEQJQN D[ Saccharomyces cerevisiae YKVJ EJTQOQUQOGÿKPVGITCVGF OWNVKEQR[ LEU4 IGPGU. $KQUEK $KQVGEJ $KQEJGO 5÷:1÷8ôÿ1÷83 *QHHOCP L üôýýýú 6JG FGHGEVKXG URQTWNCVKQP QH NCIGT DTGYKPI [GCUV. 2J& VJGUKUþ ùPKXGTUKV[ QH %QRGPJCIGPþ %QRGPJCIGP *QNODGTIþ S ü1û8ôú )GPGVKE FKHHGTGPEGU DGVYGGP Saccharomyces carlsbergensis CPF S cerevisiae ++. ñGUVTKEVKQP GPFQPWENGCUG CPCN[UKU QH IGPGU KP EJTQOQUQOG +++. %CTNUDGTI ñGU %QOOWP 47:ô33ÿô44 *QWIJ ,Sþ $TKIIU &©þ SVGXGPU ñ CPF ;QWPI 69 ü1û8ôú /CNVKPI CPF $TGYKPI SEKGPEGþ òQNWOG ++ *QRRGF 9QTV CPF $GGT. %JCROCP CPF *CNNþ LQPFQPþ ù,CEMUQP ©#þ $CNCPEG )/ CPF 6JQOUGP -- ü1û8÷ú %QPUVTWEVKQP QH C [GCUV XGEVQT FKTGEVKPI VJG U[PVJGUKU CPF TGNGCUG QH DCTNG[ ü1.3þ 1.4úÿ¾ÿINWECPCUG. %CTNUDGTI ñGU %QOOWP 51:445ÿ458 ,COGU 6%þ %CORDGNN Sþ &QPPGNN[ & CPF $QPF ù üôýý3ú 6TCPUETKRVKQP RTQHKNG QH DTGYGT[ [GCUV WPFGT HGTOGPVCVKQP EQPFKVKQPU. , #RRN /KETQDKQN û4:43ôÿ448 ,QJCPPGUGP 2( ü1ûû4ú +PETGCUKPI VJG HNWZ KP VJG UWNRJWT CUUKOKNCVQT[ RCVJYC[ õXGTRTQFWEÿ VKQP QH MET3þ MET14 CPF MET16 KP Saccharomyces cerevisiae. / SE VJGUKUþ ùPKXGTÿ UKV[ QH %QRGPJCIGPþ %QRGPJCIGP ,QJCPPGUGP 2( CPF *CPUGP , üôýýôú &KHHGTGPVKCN VTCPUETKRVKQPCN TGIWNCVKQP QH IGPG JQÿ OQGQNQIWGU KP C HWPICN URGEKGU J[DTKF. (©/S ;GCUV ñGU 1:315ÿ3ôô ,QJCPPGUGP 2(þ 0[DQTI / CPF *CPUGP , ü1ûûûú %QPUVTWEVKQP QH S carlsbergensis DTGYGT•U [GCUV YKVJQWV RTQFWEVKQP QH UWNRJKVG. 2TQE ô7VJ %QPH ©WT $TGY %QPXþ %CPPGUþ RR ÷55ÿ ÷÷ô -CYCJCVC /þ #OCTK Sþ 0KUJK\CYC ; CPF #MCFC ñ ü1ûûûú # RQUKVKXG UGNGEVKQP HQT RNCUOKF NQUU KP Saccharomyces cerevisiae WUKPI ICNCEVQUGÿKPFWEKDNG ITQYVJ KPJKDKVQT[ UGÿ qWGPEGU. ;GCUV 15:1ÿ1ý -KGNNCPFÿ$TCPFV /% ü1ûû4Cú 0GY FGXGNQROGPVU KP VJG RTQFWEVKQP QH CNEQJQNKE DGXGTCIGU ÿ OQFKHKECVKQP QH GZKUVKPI DKQEJGOKECN RCVJYC[U KP [GCUV. +P:#NDGTIJKPC Lþ (TQPVCNK L CPF SGPUK 2þ GFU:©%$÷ 2TQE ÷VJ ©WT %QPIT $KQVGEJPQN ü(NQTGPEGúþ 1ûû3 ©NUGXKGT SEKÿ GPEG $òþ #OUVGTFCO RR 1ýû1ÿ1ýû5 -KGNNCPFÿ$TCPFV /% ü1ûû4Dú +PFWUVTKCN Saccharomyces [GCUVU +P:,QJPUVQPþ ,ñþ GF:/QNGEWNCT )GPGVKEU QH ;GCUV. # 2TCEVKECN #RRTQCEJ.þ õZHQTF ùPKXGTUKV[ 2TGUUþ õZÿ HQTFþ RR ô47ÿô5û -KGNNCPFÿ$TCPFV /%þ )jGTOCPUGP %þ 0KNUUQPÿ6KNNITGP 6 CPF *QNODGTI S ü1û8ûú ;GCUV DTGGFKPI. 2TQE ôôPF %QPIT ©WT $TGY %QPXþ 1û8ûþ <·TKEJ RR 37ÿ47 -KGNNCPFÿ$TCPFV /%þ )jGTOCPUGP %þ 6WNNKP Sþ 0KNUUQPÿ6KNNITGP 6þ SKIUICCTF 2 CPF *QNOÿ DGTI S ü1ûûýú )GPGVKE CPCN[UKU CPF DTGGFKPI QH DTGYGT•U [GCUV. +P:*GUNQV *þ &CXKGU ,þ (NQTGPV ,þ $QDKEJQP Lþ &WTCPF ) CPF 2GPCUUG Lþ GFU:2TQE ÷VJ +PVGTPCVKQPCN S[ORQÿ UKWO QP )GPGVKEU QH +PFWUVTKCN /KETQQTICPKUOU.þ XQN ++þ 1ûûýþ SQEKoVo (TCPnCKUG FG /Kÿ ETQDKQNQIKGþ SVTCUDQWTIþ RR 877ÿ885
1÷÷
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
-KGNNCPFÿ$TCPFV /%þ 0KNUUQPÿ6KNNITGP 6þ )jGTOCPUGP %þ *QNODGTI S CPF 2GFGTUGP /$ ü1ûû5ú )GPGVKEU QH DTGYKPI [GCUVU. +P:9JGCNUþ #©þ ñQUGþ #* CPF *CTTKUQPþ ,Sþ GFU:6JG ;GCUVU.þ ôPF GFPþ òQN ÷ #ECFGOKE 2TGUUþ LQPFQPþ ù-þ RR ôô3ÿô54 -KO -þ $CjU\CT )þ LGG S;þ -PWFUGP (þ /CVVQQP ,ñ ü1ûû4ú %NQPKPI QH C PGY CNNGNKE XCTKCPV QH C Saccharomyces diastaticus INWEQCO[NCUG IGPG CPF KVU KPVTQFWEVKQP KPVQ KPFWUVTKCN [GCUVU. #RRN $KQEJGO $KQVGEJ 44:1÷1ÿ185 -QDC[CUJK õþ *C[CUJK 0þ -WTQMK ñ CPF SQPG * ü1ûû8ú ñGIKQP QH FLO1 RTQVGKPU TGURQPUKÿ DNG HQT UWICT TGEQIPKVKQP. , $CEVGTKQN 18ý:÷5ý3ÿ÷51ý -QFCOC ;þ (WMWK 0þ #UJKMCTK 6 CPF SJKDCPQ ; ü1ûû5ú +ORTQXGOGPV QH OCNVQUG HGTOGPVCÿ VKQP GHHKEKGPE[: %QPUVKVWVKXG GZRTGUUKQP QH MAL IGPGU KP DTGYKPI [GCUV. , #O SQE $TGY %JGO 53:ô4ÿôû -QFCOC ;þ õOWTC (þ /K[CjKOC - CPF #UJKMCTK 6 üôýý1ú %QPVTQN QH JKIJGT CNEQJQN RTQFWEÿ VKQP D[ OCPKRWNCVKQP QH VJG BAP2 IGPG KP DTGYKPI [GCUV. , #O SQE $TGY %JGO 5û:157ÿ1÷ô -QTEJ %þ /QWPVCKP *#þ )[NNCPI *þ 9KPIG / CPF $TGJOGT 2 ü1ûû1ú # OGEJCPKUO HQT UWNÿ RJKVG RTQFWEVKQP KP DGGT CPF JQY VQ KPETGCUG UWNRJKVG NGXGNU D[ TGEQODKPCPV IGPGVKEU. 2TQE ô3TF %QPIT ©WT $TGY %QPXþ 1ûû1þ LKUDQP RR ôý1ÿôý8 -WP\G )þ $QFG ñþ ñKPVCNC * CPF *QHGOGKUVGT , ü1û8ûú *GVGTQNQIQWU IGPG GZRTGUUKQP QH VJG IN[RJQUCVG TGUKUVCPEG OCTMGT CPF KVU CRRNKECVKQP KP [GCUV VTCPUHQTOCVKQP. %WTT )GPGV 15:û1ÿû8 LCPECUJKTG 9©þ %CTVGT #6þ *QYCTF ,, CPF 9KNFG ñ, ü1û8ûú SWRGTCVVGPWCVKPI DTGYKPI [GCUV. 2TQE ôôPF %QPIT ©WT $TGY %QPXþ 1û8ûþ <·TKEJ RR 4û1ÿ4û8 LGG Sþ òKNNC - CPF 2CVKPQ * ü1ûû5ú ;GCUV UVTCKP FGXGNQROGPV HQT GPJCPEGF RTQFWEVKQP QH FGUKTCDNG CNEQJQNUöGUVGTU KP DGGT. , #O SQE $TGY %JGO 53:153ÿ15÷ LGGOCPU %þ &WRKTG S CPF /CETQP ,ÿ; ü1ûû3ú ñGNCVKQP DGVYGGP YQTV &/Sõ CPF &/S EQPEGPVTCVKQP KP DGGT. 2TQE ©WT $TGY %QPX %QPITþ õUNQþ R 7ýûÿ71÷ /GCFGP 2) CPF 6WDD ñS ü1û85ú # RNCUOKF XGEVQT U[UVGO HQT VJG IGPGVKE OCPKRWNCVKQP QH DTGYKPI UVTCKPU. 2TQE ôýVJ %QPIT ©WT $TGY %QPXþ 1û85þ *GNUKPMK RR ô1ûÿôô÷ /GNFICCTF /þ *CTVJKNN ,þ 2GVGTUGP $ CPF õNUGP õ ü1ûû5ú )N[ECP OQFKHKECVKQP QH C VJGTOQÿ UVCDNG TGEQODKPCPV ü1ÿ3þ 1ÿ4úÿ¾ÿINWECPCUG UGETGVGF HTQO Saccharomyces cerevisiae KU FGVGTOKPGF D[ UVTCKP CPF EWNVWTG EQPFKVKQPU. )N[EQEQPj , 1ô:38ýÿ3ûý /KVJKGWZ S/ CPF 9GKUU #S ü1ûû5ú 6CPFGO KPVGITCVKQP QH OWNVKRNG ILV5 EQRKGU CPF GNGÿ XCVGF VTCPUETKRVKQP KP RQN[RNQKF [GCUV. ;GCUV 11:311ÿ31÷ 0CICUCYC 0þ $QICMK 6þ +YCOCVUW #þ *COCEJK / CPF -WOCICK % ü1ûû8ú %NQPKPI CPF PWÿ ENGQVKFG UGqWGPEG QH VJG CNEQJQN CEGV[NVTCPUHGTCUG ++ IGPG üATF2ú HTQO Saccharomyces cerevisiae. -[QMCK 0Q 7 $KQUEK $KQVGEJPQN $KQEJGO ÷ô:185ôÿ1857 0KNUUQPÿ6KNNITGPþ 6þ )jGTOCPUGPþ %þ -KGNNCPFÿ$TCPFVþ /%þ 2GVGTUGPþ ,)L CPF *QNODGTIþ S ü1û81ú )GPGVKE FKHHGTGPEGU DGVYGGP Saccharomyces carlsbergensis CPF S cerevisiae #PCN[UKU QH EJTQOQUQOG +++ D[ UKPING EJTQOQUQOG VTCPUHGT. %CTNUDGTI ñGU %QOOWP 4÷:÷5ÿ7÷ 0KNUUQPÿ6KNNITGP 6þ )jGTOCPUGP %þ *QNODGTI Sþ 2GVGTUGP ,)L CPF -KGNNCPFÿ$TCPFV /% ü1û8÷ú #PCN[UKU QH EJTQOQUQOG ò CPF VJG ILV1 IGPG HTQO Saccharomyces carlsbergensis. %CTNUDGTI ñGU %QOOWP 51:3ýûÿ3ô÷ 0QTFNøXþ * ü1û85ú (QTOCVKQP QH UWNRJWT FKQZKFG FWTKPI DGGT HGTOGPVCVKQP. 2TQE ôýVJ %QPIT ©WT $TGY %QPXþ 1û85þ *GNUKPMK RR ôû1ÿôû8 õNGUGP -þ ,QJCPPGUGP 2(þ *QHHOCPP Lþ SøTGPUGP S$þ )jGTOCPUGP % CPF *CPUGP , üôýýýú 6JG R;% RNCUOKFUþ C UGTKGU QH ECUUGVVGÿDCUGF [GCUV RNCUOKF XGEVQTU RTQXKFKPI OGCPU QH EQWPVGTÿUGNGEVKQP. ;GCUV 1÷:1ý35ÿ1ý43
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
1÷7
õNGUGP -þ (GNFKPI 6þ )jGTOCPUGP % CPF *CPUGP , üôýýôú 6JG F[PCOKEU QH VJG Saccharomyces carlsbergensis DTGYKPI [GCUV VTCPUETKRVQOG FWTKPI C RTQFWEVKQPÿUECNG NCIGT DGGT HGTOGPVCVKQP. (©/S ;GCUV ñGU ô:5÷3ÿ573 õOWTC (þ SJKDCPQ ;þ (WMWK 0 CPF 0CMCVCPK - ü1ûû5ú ñGFWEVKQP QH J[FTQIGP UWNRJKFG RTQÿ FWEVKQP KP DTGYKPI [GCUV D[ EQPUVKVWVKXG GZRTGUUKQP QH MET25 IGPG. , #O SQE $TGY %JGO 53:58ÿ÷ô õPQ $ÿ+þ -KjKOC -þ +UJKK 0þ -CYCVQ 6þ /CVUWFC #þ 2CU\GYUMK #þ SJKPQFC S ü1ûû÷ú ñGIWNCÿ VKQP QH UWNRJCVG CUUKOKNCVKQP KP Saccharomyces cerevisiae. ;GCUV 1ô:1153ÿ11÷ô 2CPQWVUQRQWNQW -þ 9W ,þ *C[GUþ #þ $WVNGTþ 2 CPF õNKXGTþ S) üôýý1ú ;GCUV VTCPUETKRVQOG CPCN[UKU FWTKPI VJG DTGYKPI RTQEGUU. ;GCUV 18:S3ýý 2CTM %Sþ 2CTM ;,þ LGG ;*þ 2CTM -,þ -CPI *S CPF 2GM ù* ü1ûûýú 6JG PQXGN IGPGVKE OCÿ PKRWNCVKQP VQ KORTQXG VJG RNCUOKF UVCDKNKV[ CPF GP\[OG CEVKXKV[ KP VJG TGEQODKPCPV DTGYKPI [GCUV. /$## 6GEJ íWCTV ô7:11ôÿ11÷ 2CU\GYUMK #þ õPQ $ÿ+ ü1ûûôú $KQU[PVJGUKU QH UWNRJWT COKPQ CEKFU KP Saccharomyces cerevisiae:TGIWNCVQT[ TQNGU QH OGVJKQPKPG CPF SÿCFGPQU[NOGVJKQPKPG TGCUUGUUGF. %WTT )GPGV ôô:ô73ÿô75 2GFGTUGPþ /$ ü1û85ú &0# UGqWGPEG RQN[OQTRJKUOU KP VJG IGPWU Saccharomyces ++ #PCN[UKU QH VJG IGPGU RDN1þ HIS4þ LEU2 CPF Ty VTCPURQUCDNG GNGOGPVU KP %CTNUDGTIþ 6WDQTI CPF ôô $CXCTKCP DTGYKPI UVTCKPU. %CTNUDGTI ñGU %QOOWP 5ý:ô÷3ÿô7ô 2GFGTUGP /$ ü1û8÷Cú &0# UGqWGPEG RQN[OQTRJKUO KP VJG IGPWU Saccharomyces +++ ñGÿ UVTKEVKQP GPFQPWENGCUG HTCIOGPV RCVVGTPU QH EJTQOQUQOCN TGIKQPU KP DTGYKPI CPF QVJGT [GCUV UVTCKPU. %CTNUDGTI ñGU %QOOWP 51:1÷3ÿ183 2GFGTUGP /$ ü1û8÷Dú &0# UGqWGPEG RQN[OQTRJKUO KP VJG IGPWU Saccharomyces +ò *Qÿ OQNQIQWU EJTQOQUQOGU +++ KP Saccharomyces bayanusþ S carlsbergensisþ CPF S uvarum. %CTNUDGTI ñGU %QOOWP 51:185ÿôýô 2GFGTUGPþ /$ ü1ûû4ú /QNGEWNCT CPCN[UGU QH [GCUV &0# ÿ VQQNU HQT RWTG [GCUV OCKPVGPCPEG KP VJG DTGYGT[. , #O SQE $TGY %JGO 5ô:ô3ÿô7 2GPVVKNk /þ SWKJMQ /Lþ LGJVKPGP ùþ 0KMMQNC / CPF -PQYNGU ,-% ü1û87Cú %QPUVTWEVKQP QH DTGYGT•U [GCUVU UGETGVKPI HWPICN GPFQÿ¾ÿINWECPCUGU. %WTT )GPGV 1ô:413ÿ4ôý 2GPVVKNk /þ #PFTo Lþ SCNQJGKOQ /þ LGJVQXCCTC 2þ -PQYNGU ,-% ü1û87Dú ©ZRTGUUKQP QH VYQ Trichoderma reesei GPFQINWECPCUGU KP VJG [GCUV Saccharomyces cerevisiae. ;GCUV 3:175ÿ185 2GPVVKNk / CPF ©PCTK 6ÿ/ ü1ûû1ú )GPGVKE GPIKPGGTKPI QH KPFWUVTKCN [GCUVU. +P:%JGTGOKUKPQHHþ 20 CPF (GTTCPVGþ L/ GFU:$KQVGEJPQNQI[ ÿ %WTTGPV 2TQITGUU.þ òQN 1þ 6GEJPQOKE 2WDNKUJKPI %Qþ +PEþ LCPECUVGTþ ù- RR 173ÿôýô 2GTT[ % CPF /GCFGP 2 ü1û88ú 2TQRGTVKGU QH C IGPGVKECNN[ÿGPIKPGGTGF FGZVTKPÿHGTOGPVKPI UVTCKP QH DTGYGTU• [GCUV. , +PUV $TGY û4:÷4ÿ÷7 2GVGTUGP ,)Lþ 0KNUUQPÿ6KNNITGP 6þ -KGNNCPFÿ$TCPFV /%þ )jGTOCPUGP % CPF *QNODGTI S ü1û87ú SVTWEVWTCN JGVGTQ\[IQUKU CV IGPGU ILV2 CPF ILV5 KP Saccharomyces carlsbergensis. %WTT )GPGV 1ô:1÷7ÿ174 2QNCKPC , üôýýôú $TGYGT•U ;GCUV:)GPGVKEU CPF $KQVGEJPQNQI[ +P:#RRNKGF /[EQNQI[ CPF $KQVGEJPQNQI[. òQNWOG ô:#ITKEWNVWTG CPF (QQF 2TQFWEVKQP. ©NUGXKGT SEKGPEG $òþ #OUVGTFCOþ RR 1ÿ17 2WIJ 6þ &WPP $þ òGPVGKEJGT #þ /GV\PGT Sþ $QYGT 2þ $QPFTG %þ SGCDTQQMU ,þ ñ[FGT &þ $QVUVGKP & CPF $TQYP 2 üôýýôú )NQDCN CPCN[UKU QH [GCUV IGPG GZRTGUUKQP FWTKPI C DTGYGT[ HGTOGPVCVKQP. +P: #PPWCN /GGVKPI QH VJG #OGTKECP SQEKGV[ QH $TGYKPI %JGOÿ KUVUþ ôýýôþ #DUVTCEVUþ Qô
1÷8
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
ñGUPKEMþ /#þ SMCCPKNFþ / CPF 0KNUUQPÿ6KNNITGPþ 6 ü1û8ûú LCEM QH &0# JQOQNQI[ KP C RCKT QH FKXGTIGPV EJTQOQUQOGU ITGCVN[ UGPUKVK\GU VJGO VQ NQUU D[ &0# FCOCIG. 2TQE 0CVN #ECF SEK ùS# 8÷:ôô7÷ÿôô8ý ñQOCPQU /#þ SEQTGT %# CPF %NCTG ,, ü1ûûôú (QTGKIP IGPG GZRTGUUKQP KP [GCUV:# TGXKGY. ;GCUV 8:4ô3ÿ488 ñQVJUVGKP ñ ü1ûû1ú 6CTIGVVKPIþ &KUTWRVKQPþ ñGRNCEGOGPVþ CPF CNNGNG TGUEWG:+PVGITCVKXG &0# VTCPUHQTOCVKQP KP [GCUV. /GVJ ©P\[OQN 1û4:ô81ÿ3ý1 ñWUUGNN + CPF SVGYCTV )) ü1û7ûú SRJGTQRNCUV HWUKQP QH DTGYGT•U [GCUV UVTCKPU. , +PUV $TGY 85:û5ÿû8 SCMCK -þ (WMWK Sþ ;CDWWEJK Sþ #Q[CIK S CPF 6UWOWTC ; ü1û8ûú ©ZRTGUUKQP QH VJG Saccharomyces diastaticus STA1 IGPG KP DTGYKPI [GCUVU. , #O SQE $TGY %JGO 47:87ÿû1 SCOWGN & ü1ûû÷ú #TEJCGQNQI[ QH CPEKGPV ©I[RVKCP DGGT. , #O SQE $TGY %JGO 54:3ÿ1ô SCVC[CPCTC[CPC 6þ ùODCTIGT *© CPF LKPFGITGP ) ü1û÷8ú $KQU[PVJGUKU QH DTCPEJGFÿEJCKP COKPQ CEKFU KP [GCUV:ñGIWNCVKQP QH NGWEKPG DKQU[PVJGUKU KP RTQVQVTQRJKE CPF NGWEKPG CWZQVTQRJKE UVTCKPU. , $CEVGTKQN û÷:ôý18ÿôýô4 SEJGTGT S CPF &CXKU ñ9 ü1û7ûú ñGRNCEGOGPV QH EJTQOQUQOG UGIOGPVU YKVJ CNVGTGF &0# UGqWGPEGU EQPUVTWEVGF in vitro. 2TQE 0CVN #ECF SEK ùS# 7÷:4û51ÿ4û55 SJKOK\W (þ SQPG * CPF +PQWG 6 ü1û8ûú $TGYKPI RGTHQTOCPEG QH C IGPGVKECNN[ VTCPUHQTOGF [GCUV YKVJ CEGVQNCEVCVG FGECTDQZ[NCUG CEVKXKV[. /$## 6GEJ íWCTV ô÷:47ÿ5ý SQPG *þ -QPFQ -þ (WjKK 6þ SJKOW\W (þ 6CPCMC , CPF +PQWG 6 ü1û87ú (GTOGPVCVKQP RTQRGTÿ VKGU QH DTGYGT•U [GCUV JCXKPI "ÿCEGVQNCEVCVG FGECTDQZ[NCUG IGPG. 2TQE ô1UV %QPIT ©WT $TGY %QPXþ 1û87þ /CFTKF RR 545ÿ55ô SVGYCTV ))þ )CTTKUQP +(þ )QTKPI 6©þ /GNGI /þ 2KRCUVU 2 CPF ñWUUGNN + ü1û7÷ú $KQEJGOKECN CPF IGPGVKE UVWFKGU QP [GCUV HNQEEWNCVKQP. -GOKCÿ-GOK ü*GNUKPMKú 3:4÷5ÿ47û SVGYCTV ))þ ñWUUGNN + CPF SKNNU #/ ü1û83ú (CEVQTU VJCV EQPVTQN VJG WVKNK\CVKQP QH YQTV ECTÿ DQJ[FTCVGU D[ [GCUV. /$## 6GEJ íWCTV ôý:1ÿ8 SVGYCTV ))þ ,QPGU ñ CPF ñWUUGNN + ü1û85ú 6JG WUG QH FGTGRTGUUGF [GCUV OWVCPVU KP VJG HGTÿ OGPVCVKQP QH DTGYGT[ YQTV. 2TQE ôýVJ %QPIT ©WT $TGY %QPXþ 1û85þ *GNUKPMK RR ô43ÿ ô5ý SVTCVHQTF / ü1ûûôú ;GCUV HNQEEWNCVKQP: TGEQPEKNKCVKQP QH RJ[UKQNQIKECN CPF IGPGVKE XKGYÿ RQKPVU. ;GCUV 8:ô5ÿ38 SWKJMQ /ÿLþ $NQOqWKUV -þ 2GPVVKNk /þ )KUNGT ñ CPF -PQYNGU , ü1ûûýú ñGEQODKPCPV DTGYGT•U [GCUV UVTCKPU UWKVCDNG HQT CEEGNGTCVGF DTGYKPI. , $KQVGEJPQN 14:ô85ÿ3ýý 6CFC Sþ 6CMGWEJK 6þ SQPG *þ ;COCPQ Sþ SEJQHKGNF /#þ *COOQPF ,ñ/ CPF +PQWG 6 ü1ûû5ú 2KNQVÿUECNG DTGYKPI YKVJ KPFWUVTKCN [GCUVU YJKEJ RTQFWEG VJG "ÿCEGVQNCEVCVG FGÿ ECTDQZ[NCUG QH Acetobacter aceti UUR xylinum. 2TQE ô5TF %QPIT ©WT $TGY %QPXþ 1ûû5þ $TWUUGNUþ RR 3÷ûÿ37÷ 6COCK ;þ /QOOC 6þ ;QUJKOQVQ * CPF -CPGMQ ; ü1ûû8ú %QÿGZKUVGPEG QH VYQ V[RGU QH EJTQOQUQOG KP VJG DQVVQO HGTOGPVKPI [GCUVþ Saccharomyces pastorianus. ;GCUV 14:ûô3ÿû33 6COCK ;þ 6CPCMC -þ ùOGOQVQ 0þ 6QOK\WMC - CPF -CPGMQ ; üôýýýú &KXGTUKV[ QH VJG *õ IGPG GPEQFKPI CP GPFQPWENGCUG HQT OCVKPI V[RG EQPXGTUKQP KP VJG DQVVQO HGTOGPVKPI [GCUV Saccharomyces pastorianus. ;GCUV 1÷:1335ÿ1343 6GWPKUUGP #9ñ*þ *QNWD ©þ XCP FGT *WEJV ,þ XCP FGP $GTI ,# CPF SVGGPUOC *; ü1ûû3ú SGqWGPEG QH VJG QRGP TGCFKPI HTCOG QH VJG FLO1 IGPG HTQO Saccharomyces cerevisiae. ;GCUV û:4ô3ÿ4ô7
5 $TGYGT•U [GCUV: IGPGVKE UVTWEVWTG CPF VCTIGVU HQT KORTQXGOGPV
1÷û
6GWPKUUGP #9ñ* CPF SVGGPUOC *; ü1ûû5ú ñGXKGY: VJG FQOKPCPV HNQEEWNCVKQP IGPGU QH Saccharomyces cerevisiae EQPUVKVWVG C PGY UWDVGNQOGTKE IGPG HCOKN[. ;GCUV 11:1ýý1ÿ 1ý13 6G\WMC *þ /QTK 6þ õMWOWTC ;þ -KVCDCVCMG - CPF 6UWOWTC ; ü1ûûôú %NQPKPI QH C IGPG UWRRTGUUKPI J[FTQIGP UWNRJKFG RTQFWEVKQP D[ Saccharomyces cerevisiae CPF KVU GZÿ RTGUUKQP KP C DTGYKPI [GCUV. , #O SQE $TGY %JGO 5ý:13ýÿ133 6WDD ñSþ SGCTNG $#þ )QQFG[ #ñ CPF $TQYP #,2 ü1û81ú ñCTG OCVKPI CPF VTCPUHQTOCVKQP HQT EQPUVTWEVKQP QH PQXGN DTGYKPI [GCUVU. 2TQE 18VJ %QPIT ©WT $TGY %QPXþ 1û81þ %Qÿ RGPJCIGP RR 487ÿ4û÷ ùTCPQ 0þ SCJCTC * CPF -QUJKPQ S ü1ûû3Cú %QPXGTUKQP QH C PQPÿHNQEEWNGPV DTGYGT•U [GCUV VQ HNQEEWNGPV QPGU D[ GNGEVTQHWUKQP 1. +FGPVKHKECVKQP CPF EJCTCEVGTK\CVKQP QH VJG HWUCPVU D[ RWNUGF HKGNF IGN GNEVTQRJQTGUKU. , $KQVGEJPQN ô8:ô37ÿô47 ùTCPQ 0þ SCVQ /þ SCJCTC * CPF -QUJKPQ S ü1ûû3Dú %QPXGTUKQP QH C PQPÿHNQEEWNGPV DTGYGT•U [GCUV VQ HNQEEWNGPV QPGU D[ GNGEVTQHWUKQP ô. SOCNNÿUECNG DTGYKPI D[ HWUCPVU, $KQVGEJPQN ô8:ô4ûÿô÷1 òCMGTKC & CPF *KPEJNKHHG © ü1û8ûú #O[NQN[VKE DTGYKPI [GCUV:VJGKT EQOOGTEKCN CPF NGIKUNCÿ VKXG CEEGRVCDKNKV[. 2TQE ôôPF %QPIT ©WT $TGY %QPXþ 1û8ûþ <·TKEJ RR 475ÿ48ô òCMGTKC &þ $QZ 9þ $KTF L CPF /GNNQT , ü1ûû÷ú %JCTCEVGTKUCVKQP QH CO[NQN[VKE DTGYKPI [GCUV. , +PUV $TGY 1ýô:ô7ÿ3ô òCWIJCPÿ/CTVKPK # CPF -WTV\OCP %2 ü1û85ú &GQZ[TKDQPWENGKE CEKF TGNCVGFPGUU COQPI URGEKGU QH Sacharomyces sensu stricto. +PV , S[UV $CEVGTKQN 35:5ý8ÿ511 òCWIJCPÿ/CTVKPK # CPF /CTVKPK # ü1û87ú 6JTGG PGYN[ FGNKOKVGF URGEKGU QH Saccharomyces sensu stricto. #PVQPKG X LGGWYGPJQGM 53:77ÿ84 òGTUVTGRGP -þ $CWGT (þ /KEJKGNU %þ >FGNKPEMZ )þ &GNXCWZ ( CPF 2TGVQTKWU + üôýýýú KP:©$% /QPQITCRJ ô8þ ©$%ÿS[ORQUKWO ;GCUV 2J[UKQNQI[ # 0GY ©TC QH õRRQTVWÿ PKV[. (CEJXGTNCI *CPU %CTNþ 0·TPDGTIþ RR. 3ýÿ4ô òKNNCPWGDC -&þ )QQUUGPU © CPF /CUUEJGNGKP %# ü1ûûýú SWDVJTGUJQNF XKEKPCN FKMGVQPG NGXGNU KP NCIGT DTGYKPI [GCUV HGTOGPVCVKQPU D[ OGCPU QH ILV5 IGPG CORNKHKECVKQP. , #O SQE $TGY %JGO 48:111ÿ114 9CNMGT /& CPF SKORUQP 9, ü1ûû3ú 2TQFWEVKQP QH XQNCVKNG UWNRJWT EQORQWPFU D[ CNG CPF NCIGT DTGYKPI UVTCKPU QH SCEEJCTQO[EGU EGTGXKUKCG. LGVV #RRN /KETQDKQN 1÷:4ýÿ43 9CVCPCDG /þ 6CPCMC 0þ /KUJKOC * CPF 6CMGOWTC S ü1ûû3ú +UQNCVKQP QH UCMG [GCUV OWÿ VCPVU TGUKUVCPV VQ KUQCO[N OQPQHNWQTQCEGVCVG VQ KORTQXG KUQCO[N CEGVCVG RTQFWEVKXKV[. , (GTO $KQGPI 7÷:ôôûÿô31 9CVCTK ,þ 6CMCVC ;þ õICYC /þ SCJCTC *þ -QUJKPQ Sþ õPPGNC /Lþ #KTCMUKPGP ùþ ,CCVKPGP ñþ 2GPVVKNk / CPF -GTkPGP S ü1ûû4ú /QNGEWNCT ENQPKPI CPF CPCN[UKU QH VJG [GCUV HNQEEWNCÿ VKQP IGPG FLO1. ;GCUV 11:ô11ÿôô5 ;COCIKUJK * CPF õICVC 6 ü1ûûûú %JTQOQUQOCN UVTWEVWTGU QH DQVVQO HGTOGPVKPI [GCUVU. S[UV #RRN /KETQDKQN ôô:341ÿ353 ;COCPQ Sþ 6CPCMC , CPF +PQWG 6 ü1ûû4Cú %NQPKPI CPF GZRTGUUKQP QH VJG IGPG GPEQFKPI "ÿ CEGVQNCEVCVG FGECTDQZ[NCUG HTQO Acetobacter acetii UUR xylinum KP DTGYGT•U [GCUV. , $KQVGEJPQN 3ô:1÷5ÿ171 ;COCPQ Sþ -QPFQ -þ 6CPCMC , CPF +PQWG 6 ü1ûû4Dú %QPUVTWEVKQP QH C DTGYGT•U [GCUV JCXÿ KPI "ÿCEGVQNCEVCVG FGECTDQZ[NCUG IGPG HTQO Acetobacter acetii UUR xylinum KPVGITCVGF KP VJG IGPQOG , $KQVGEJPQN 3ô:173ÿ178 ;COCPQ Sþ 6QOK\WMC -þ SQPG *þ +OWTC /þ 6CMGWEJK 6þ 6CPCMC , CPF +PQWG 6 ü1ûû5ú $TGYKPI RGTHQTOCPEG QH C DTGYGT•U [GCUV JCXKPI "ÿCEGVQNCEVCVG FGECTDQZ[NCUG HTQO Acetobacter aceti UUR xylinum. , $KQVGEJPQN 3û:ô1ÿô÷
17ý
,øTIGP *CPUGP CPF /QTVGP %. -KGNNCPFÿ$TCPFV
;CTTQYþ & ü1û84ú SCEEJCTQO[EGU /G[GP GZ ñGGUU +P:0,9 -TGIGTÿXCP ñKjþ GF:6JG ;GCUVUþ C 6CZQPQOKE SVWF[.þ 3TF GFPþ ©NUGXKGT SEKGPEG 2WDNKUJGTUþ #OUVGTFCOþ RR 37ûÿ3û5 ;QEWOþ ññ ü1û85ú ©WTQRGCP 2CVGPV 0Q 853ý3÷ô5û ;QEWO ññ ü1û8÷ú )GPGVKE GPIKPGGTKPI QH KPFWUVTKCN [GCUVU. 2TQE $KQ ©ZRQ 8÷þ 1û8÷þ $QUÿ VQP RR 171ÿ18ý ;QUJKOQVQ *þ (WjKYCTC &þ /QOOC 6þ 6CPCMC -þ SQPG *þ 0CICUCYC 0 CPF 6COCK ; ü1ûûûú +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH VJG ATF2 IGPG GPEQFKPI CNEQJQN CEGV[NVTCPUÿ HGTCUG ++ KP VJG DQVVQO HGTOGPVKPI [GCUV Saccharomyces pastorianus. ;GCUV 15:4ýûÿ417
6 Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP -CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
#DUVTCEV (QT Kluyveromyces lactis DCUKE TGUGCTEJ, FGXGNQROGPV QH IGPGVKE VQQNU CPF KPFWUÿ VTKCN CRRNKECVKQP JCU IQPG JCPF KP JCPF QXGT VJG NCUV HKHVGGP [GCTU. 6JKU CTVKENG UWOOCTK\GU VJG EWTTGPV MPQYNGFIG YKVJ RCTVKEWNCT GORJCUKU QP IGPGVKEU CPF RJ[UKQNQI[ KP EQORCTKUQP VQ Saccharomyces cerevisiae. 6JG K. lactis IGPQOG UGÿ SWGPEKPI RTQLGEV KU ENQUG VQ EQORNGVKQP CPF JCU CNTGCF[ UJQYP VJCV, KP EQPVTCUV VQ S. cerevisiae, KPVGPUKXG IGPG FWRNKECVKQPU CTG NCEMKPI YJGTGCU C TGRGTVQKTG QH IGPGU PQV RTGUGPV KP S. cerevisiae EQPHGTU C DTQCFGT UWDUVTCVG URGEVTWO. /CTMGF FKHHGTÿ GPEGU GZKUV YKVJ TGURGEV VQ TGIWNCVKQP QH ECTDQP OGVCDQNKUO CPF VJG DCNCPEG DGÿ VYGGP HGTOGPVCVKQP CPF TGURKTCVKQP. K. lactis RTQXGF UWKVCDNG CU C JQUV HQT TGEQOÿ DKPCPV RTQVGKP RTQFWEVKQP FWG VQ NQY RTQVGCUG NGXGNU, JKIJ HQNFKPI CPF UGETGVKQP ECRCEKV[, CPF VJG CDUGPEG QH C %TCDVTGG GHHGEV. )GPGVKE CPCN[UKU CPF IGPGVKE OQFKÿ HKECVKQP ECP DG RGTHQTOGF CU KP S. cerevisiae CPF VJG CXCKNCDNG VQQNU CTG IGVVKPI KPÿ ETGCUKPIN[ GNCDQTCVG.
6.1 +PVTQFWEVKQP +PFWUVTKCN KPVGTGUV KP Kluyveromyces lactis KPKVKCNN[ HQEWUUGF QP GP\[OGU, KP RCTÿ VKEWNCT NCEVCUG, YJKEJ YGTG RWTKHKGF HTQO VJKU [GCUV CPF WUGF KP DKQEQPXGTUKQP RTQEGUUGU. 6JWU, HGTOGPVCVKQP VGEJPQNQI[ KU YGNN GUVCDNKUJGF CPF VJG )4#5 þgGPÿ GTCNN[ rGICTFGF aU sCHGý UVCVWU YCU CRRTQXGF VQ GP\[OGU RTGRCTCVKQPU HTQO K. lactis CNNQYKPI VJGO VQ DG WUGF CU HQQF CFFKVKXGU. #OQPI VJQUG CTG TGEQODKPCPV GPÿ \[OGU HTQO IGPGVKECNN[ GPIKPGGTGF UVTCKPU VJCV JCXG DGEQOG CXCKNCDNG CHVGT C DCVVGT[ QH IGPGVKE VQQNU JCF DGGP FGXGNQRGF. 0QYCFC[U, K. lactis DGNQPIU VQ VJG DGUVÿGUVCDNKUJGF PQPÿSaccharomyces [GCUVU HQT YJKEJ DCUKE CU YGNN CU CRRNKGF TGÿ UGCTEJ KU IQKPI JCPF KP JCPF. 1XGT VJG RCUV HKHVGGP [GCTU, Kluyveromyces TGUGCTEJ JCU DGPGHKVGF HTQO RWDNKE HWPFKPI CPF C UOCNN DWV CEVKXGN[ KPVGTCEVKPI UEKGPVKHKE EQOOWPKV[ VJCV OGGVU QPEG C [GCT CV XCT[KPI NQECVKQPU YKVJKP ©WTQRG. 9G YKUJ VQ FGFKECVG VJKU TGXKGY CTVKENG VQ *KTQUJK (WMWJCTC YJQ YCU C HQWPFGT CPF EGPVTCN OQVQT QH VJKU EQOOWPKV[ CPF YJQ UVKNN KU C XGT[ CEVKXG UWRRQTVGT QH Kluyveromyces TGUGCTEJ. 6JCPMU VQ /QPKSWG $QNQVKPÿ(WMWJCTC þ2CTKUý YJQ OCPCIGF VQ OCMG TGCN VJG K. lactis IGPQOG UGÿ SWGPEKPI RTQLGEV CV )oPQUEQRG þ(TCPEGý. K. lactis TGUGCTEJ YKNN UQQP GPVGT VJG RQUVÿIGPQOKE GTC, YJKEJ WPFQWDVGFN[, D[ EQORCTKUQPU YKVJ VJG TGHGTGPEG [GCUV S. Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
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-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
cerevisiae, YKNN RTQXKFG XCNWCDNG KPUKIJV KPVQ IGPQOG GXQNWVKQP, OQNGEWNCT RJ[UKÿ QNQI[, CPF IGPGVKEU. # PWODGT QH TGXKGY CTVKENGU QP K. lactis JCXG DGGP RWDNKUJGF TGEGPVN[ þ9oUQNQYUMKÿúQWXGN GV CN. üùùø÷ $TGWPKI GV CN. ûööö÷ 5EJCHHTCVJ CPF $TGWPKI, ûööö÷ $QNQVKPÿ(WMWJCTC GV CN. ûöööý CPF C OCPWCN QP GZRGTKOGPVCN OGVJQFU KP K. lactis TGUGCTEJ KU CXCKNCDNG þ9QNH GV CN. ûöö3ý. 6JKU EJCRVGT YKNN VJGTGHQTG HQEWU QP VJG OQUV TGEGPV NKVGTCVWTG GORJCUK\KPI KP RCTVKEWNCT FKHHGTGPEGU DGVYGGP K. lactis CPF S. cerevisiae.
6.2 )GPGVKEU 6.2.1 ÿCZQPQO[ QH Kluyveromyces UUR. CPF RJ[NQIGPGVKE TGNCVKQPUJKR VQ S. cerevisiae 6CZQPQOKE ENCUUKHKECVKQP QH Kluyveromyces species JCU C JKUVQT[ QH EJCPIGU. Kluyveromyces lactis DCUGF QP KVU CDKNKV[ VQ WVKNKUG NCEVQUG YCU KPKVKCNN[ ENCUUKHKGF CU Saccharomyces lactis. úCVGT QP, RJ[UKQNQIKECN ETKVGTKC, NKMG VJG RGVKVGÿPGICVKXG EJCTCEVGT, VJG %TCDVTGG GHHGEV, CPF VJG KPCDKNKV[ VQ ITQY CPCGTQDKECNN[, UGRCTCVGF VJG IGPWU Kluyveromyces HTQO Saccharomyces. /QNGEWNCT V[RKPI VGEJPKSWGU HKÿ PCNN[ UGV Kluyveromyces lactis CRCTV HTQO K. marxianus CPF K. fragilis KPVQ C UGRCÿ TCVG URGEKGU þ8CWIJCPÿ/CTVKPK GV CN. üùù3÷ -WTV\OCP CPF 4QDPGVV üùùõý. 6JG HKGNFU QH OKETQDKCN VCZQPQO[ JCXG DGGP TGXQNWVKQPK\GF UKPEG IGPQOKE UGÿ SWGPEGU DGECOG CXCKNCDNG. 6JG EQORNGVKQP QH VJG S. cerevisiae IGPQOG UGSWGPEG KP üùùø RTQXKFGF CP KPXCNWCDNG CPEJQT RQKPV HQT EQORCTCVKXG HWPICN IGPQOKEU CPF GWMCT[QVKE GXQNWVKQP. Kluyveromyces lactis CPF KVU TGNCVKXGU DGNQPI VQ C ITQWR HQT YJKEJ EQORCTCVKXG CRRTQCEJGU CTG RCTVKEWNCTN[ KPHQTOCVKXG. (KTUV KPUKIJV KPVQ VJG K. lactis IGPQOG QTICPKUCVKQP YCU RTQXKFGF D[ 1\KGTÿ-CNQIGTQRQWNQU GV CN. YJQ UGSWGPEGF ôôõ TCPFQO UGSWGPEG VCIU þ456Uý þ1\KGTÿ-CNQIGTQRQWNQU GV CN. üùùõý. 6JKU CRRTQCEJ YCU GZVGPFGF KP VJG )GPQNGXWTG + RTQLGEV, C HWPICN IGPQOG UGÿ SWGPEKPI KPKVKCVKXG QH GKIJV (TGPEJ TGUGCTEJ KPUVKVWVKQPU, KP YJKEJ ü3 JGOKCUEGVQÿ O[EVQWU [GCUVU KPENWFKPI K. lactis YGTG CPCN[UGF þ5QWEKGV GV CN. ûööö÷ úNQTGPVG GV CN. ûöööC÷ úNQTGPVG GV CN. ûöööD÷ $QNQVKPÿ(WMWJCTC GV CN. ûöööý. #DQWV ûôöö 14(U JCXG DGGP KFGPVKHKGF QH YJKEJ ûô GPEQFG PGY RTQVGKPU YKVJ PQ QTVJQNQIWGU KP S. cerevisiae þ$QNQVKPÿ(WMWJCTC GV CN. ûöööý. /QUV QH VJGUG OC[ TGRTGUGPV GP\[OGU KPXQNXGF KP OGVCDQNKE RCVJYC[U CNNQYKPI K. lactis VQ CUUKOKNCVG ECTDQP CPF PKVTQÿ IGP HTQO UQWTEGU VJCV CTG PQV WUGF D[ S. cerevisiae. +PVTQPU CTG UECTEG CU KP S. cerevisiae; UGSWGPEGU YKVJ UKOKNCTKV[ VQ VTCPURQUQPU 6[ü CPF 6[û JCXG DGGP HQWPF QP LWUV C HGY ©56U. 6JGTG KU C TGOCTMCDNG UKOKNCTKV[ KP IGPG QTFGT þU[PVGP[ý DGVYGGP VJG K. lactis CPF VJG S. cerevisiae IGPQOGU þ1\KGTÿ-CNQIGTQRQWNQU GV CN. üùùõ÷ -GQIJ GV CN. üùùõý. #DQWV ôùó QH CFLCEGPV IGPG RCKTU KP K. lactis QT K. marxianus CTG CNUQ CFÿ LCEGPV KP S. cerevisiae, CPF C HWTVJGT üøó QH Kluyveromyces PGKIJDQWTU ECP DG GZÿ RNCKPGF KP VGTOU QH VJG KPHGTTGF CPEGUVTCN IGPG QTFGT KP Saccharomyces RTKQT VQ VJG QEEWTTGPEG QH CP CPEKGPV YJQNGÿIGPQOG FWRNKECVKQP. /QTGQXGT, GZVGPUKXG IGPG
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
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FWRNKECVKQPU CTG PQV HQWPF KP VJG K. lactis IGPQOG FGURKVG VJG HCEV VJCV UQOG UKPÿ INGÿEQR[ IGPGU QH S. cerevisiae JCXG FWRNKECVGU KP K. lactis þ$QNQVKPÿ(WMWJCTC GV CN. ûöööý. 6JGUG UGSWGPEG GXCNWCVKQPU JCXG NGCF VQ VJG UWIIGUVKQP VJCV K. lactis FKÿ XGTIGF HTQO VJG S. cerevisiae NKPGCIG DGHQTG VJG RTQRQUGF IGPQOG FWRNKECVKQP QEÿ EWTTGF CPF OC[ VJWU DG ENQUGT VQ VJG VTWPM QH VJG RJ[NQIGPGVKE VTGG þ9QNHG CPF 5JKGNFU üùù7÷ -GQIJ GV CN. üùùõ÷ 5GQKIJG CPF 9QNHG üùùù÷ 2KUMWT ûööüý. /QTG FGHKPKVG RTQQH KU GZRGEVGF HTQO VJG )oPQNGXWTG ++ RTQLGEV EWTTGPVN[ KP RTQITGUU, YJKEJ KU GZRGEVGF VQ [KGNF VJG EQORNGVG K. lactis IGPQOG UGSWGPEG DGHQTG ûööò. 6.2.2 %JTQOQUQOGU CPF GZVTCEJTQOQUQOCN IGPGVKE GNGOGPVU 6.2.2.1 Centromeres and telomeres on the six -. NCEVKU chromosomes 6JG JCRNQKF IGPQOG QH K. lactis EQPUKUVU QH UKZ EJTQOQUQOGU TCPIKPI KP UK\G HTQO ü.ü þHQT VJG UOCNNGUV EJTQOQUQOG +ý VQ û.ø /DR þ/.$QNQVKPÿ(WMWJCTC, RGTUQPCN EQOOWPKECVKQPý. 6JG HQWTVJ EJTQOQUQOG EQPVCKPU VJG T&0# TGRGCVU CPF CRRGCTU UQOGVKOGU FKHHWUG CPF RQQTN[ UGRCTCVGF HTQO EJTQOQUQOG 8 KP RWNUGÿHKGNF IGN GNGEVTQRJQTGUKU. # PWODGT QH IGPGU OCRRGF D[ ENCUUKECN IGPGVKE OGVJQFU YGTG CUÿ UKIPGF VQ EJTQOQUQOGU D[ J[DTKFKUCVKQP þ9oUQNQYUMKÿúQWXGN CPF (WMWJCTC üùùôý. # OQTG GZVGPUKXG OCRRKPI QH $#% ENQPGU VQ KPFKXKFWCN EJTQOQUQOGU JCU TGEGPVN[ DGGP HKPKUJGF CU RCTV QH VJG )oPQNGXWTG ++ RTQLGEV. (KXG QH VJG UKZ EGPVTQOGTGU JCXG DGGP ENQPGF D[ VJGKT CDKNKV[ VQ EQPHGT UVCDNG VTCPUOKUUKQP VJTQWIJ OKVQUKU QH #45 EQPVCKPKPI &0# CU NQY EQR[ GRKUQOGU. %GPÿ VTQOGTKE XGEVQTU JCXG DGGP FGTKXGF HTQO VJG ENQPGF K. lactis EGPVTQOGTGU CPF UJQYP VQ DG UVCDN[ OCKPVCKPGF KP üÿû EQRKGU RGT EGNN. 6JG EGPVTQOGTGU CTG QTICPÿ KUGF KP C UKOKNCT YC[ CU KP S. cerevisiae CPF EQPVCKP VJG EQPUGTXGF GNGOGPVU %&©+, %&©++, CPF %&©+++ . 6JG #6ÿTKEJ %&©++ KU CDQWV VYKEG CU NQPI CU KP S. cerevisiae CPF VJWU VJG UK\G QH VJG EGPVTQOGTGU KU CDQWV ûôö DR. &GURKVG UKOKNCT QTICPKUCVKQP, K. lactis EGPVTQOGTGU CTG WPCDNG VQ EQPHGT EGPVTQOGTG HWPEVKQP KP S. cerevisiae CPF vice versa þ*GWU GV CN. üùùò CPF TGHGTGPEGU VJGTGKPý. 5RGEKGU URGEKHKEKV[ CRRGCTU VQ DG C EQOOQP HGCVWTG QH EGPVTQOGTG HWPEVKQP GXGP HQT ENQUGN[ TGNCVGF URGEKGU. 4Gÿ OCTMCDN[, EGPVTQOGTGU HTQO Kluyveromyces marxianus YGTG UJQYP VQ HWPEVKQP KP K. lactis þ+DQTTC CPF $CNN üùùòý. 6JG OQNGEWNCT DCUKU HQT URGEKGU URGEKHKEKV[ JCU PQV DGGP GNWEKFCVGF. 6JG NGPIVJ FKHHGTGPEG KP %&©++ KU WPNKMGN[ VQ DG TGURQPUKDNG þ*GWU GV CN. üùùò ý. *QYGXGT, VJGTG KU OWEJ TQQO HQT URGEKHKEKV[ KP VJG NCTIG RTQVGKP EQORNGZ VJCV HQTOU GXGP QP VJG UOCNN EGPVTQOGTGU QH DWFFKPI [GCUVU þ%NCTMG üùùõ÷ -KVCICYC CPF *KGVGT ûööüý. 6JG %&©+++ DKPFKPI EQORNGZ YJKEJ PWENGCVGU VJG MKPGVQEJQTG QH S. cerevisiae EQPUKUVU QH 0FEüöñ%DHû, %GR3, %VHü3, CPF 5MRü. KlSKP1 YCU ENQPGF D[ UWRRTGUUKQP QH C S. cerevisiae ctf13-ts OWVCPV CPF UJQYP VQ DG CP GUUGPVKCN IGPG, CU GZRGEVGF þ9KPMNGT GV CN. ûöööCý. #VVGORVU VQ KUQNCVG C NDC10/CBF2 JQOQÿ NQIWG D[ EQORNGOGPVCVKQP QH C cbf2-ts OWVCPV HCKNGF. +PUVGCF, OWNVKÿEQR[ UWRRTGUÿ UQTU GPEQFKPI WDKSWKVKP EQPLWICVKPI CPF WDKSWKVKP J[FTQN[UKPI GP\[OGU YGTG HQWPF þ9KPMNGT GV CN. ûöööDý. KlCBF1 GPEQFKPI VJG %&©+ DKPFKPI EQORQPGPV YCU CNUQ UJQYP VQ EQORNGOGPV VJG EQTTGURQPFKPI S. cerevisiae OWVCVKQP CPF vice
ü7ò
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
versa þ/WNFGT GV CN. üùù3÷ 9KPMNGT GV CN. ûöööC÷ 9KPMNGT GV CN. ûöööDý. CBF1 KU CP GUUGPVKCN IGPG KP K. lactis DWV PQV KP S. cerevisiae þ/WNFGT GV CN. üùù3ý. +V KU RQUUKÿ DNG VJCV VJG GUUGPVKCN HWPEVKQP KP K. lactis KU PQV TGNCVGF VQ EGPVTQOGTG HWPEVKQP DWV VQ UQOG QVJGT CURGEV QH VJG OWNVKHWPEVKQPCN %DHü RTQVGKP, YJKEJ KU CNUQ KPXQNXGF KP IGPG TGIWNCVKQP þ/GNNQT GV CN. üùùö÷ /WNFGT GV CN. üùùôDý. 6GNQOGTGU, VJG PCVWTCN GPFU QH GWMCT[QVKE EJTQOQUQOGU, CTG GUUGPVKCN HQT VJG RTQVGEVKQP QH EJTQOQUQOGU HTQO GPFÿVQÿGPF HWUKQP. 6JG K. lactis VGNQOGTGU JCXG DGGP CPCN[UGF GZVGPUKXGN[ KP VJG NCD QH ©NK\CDGVJ $NCEMDWTP þ/E©CEJGTP GV CN. ûöööý. +P EQPVTCUV VQ VJG KTTGIWNCT S. cerevisiae TGRGCVU, VJG[ EQPUKUV QH üô VCPFGO EQRKGU QP CXGTCIG QH C RGTHGEV ûôÿDR TGRGCV VJCV EQPVCKPU VJG üû DR EQPUGPUWU UGÿ SWGPEG HQT 4CRü, C EQPUGTXGF VGNQOGTG DKPFKPI HCEVQT þ(KI. üý. 5'-)#66#))6#6)6))6)6#%))#666-3' 3'----###%7##7%%#7#%#%%#%#7)%%7###%7-----5' (KI. 1. K. lactis 6GNQOGTKE 4GRGCV. 6JG ûô DR VGNQOGTKE TGRGCV KU UJQYP KP VJG WRRGT NKPG YKVJ VJG 4CRü DKPFKPI UKVG KP KVCNKEU. 6JG NQYGT NKPG TGRTGUGPVU VJG EQFKPI RCTV QH VGNQOGTKE 40# GPEQFGF D[ VJG TER1 IGPG, WPFGTNKPGF CTG HNCPMKPI FKTGEV TGRGCVU. #HVGT /E©CEJGTP GV CN. þûööûý.
6JG WPWUWCNN[ NQPI ûôÿDR TGRGCV QHHGTGF VJG RQUUKDKNKV[ VQ UGRCTCVG 4CRü DKPFÿ KPI HWPEVKQP HTQO HNCPMKPI GHHGEVU. $[ OWVCIGPGUKU QH VJG TER1 IGPG GPEQFKPI VJG 40# RQTVKQP QH VGNQOGTCUG VJG TGRGCV UGSWGPEG JCU DGGP CNVGTGF CPF VWTPQXGT QH VGNQOGTKE UGSWGPEGU YCU GZCOKPGF. 6JG TGUWNVU UWRRQTVGF VJG ETWEKCN TQNG QH 4CRü KP VGNQOGTG OCKPVGPCPEG CPF TGRGCV EQR[ PWODGT EQPVTQN þ/E©CEJGTP GV CN. ûööö÷ /E©CEJGTP GV CN. ûööûý. 6.2.2.2 Linear cytoplasmic DNA plasmids conferring the Killer phenotype +P UGXGTCN K. lactis UVTCKPU, VYQ FQWDNGÿUVTCPFGF NKPGCT &0# RNCUOKFU JCXG DGGP FGVGEVGF VJCV EQPHGT C ÉMKNNGTÌ RJGPQV[RG þ)WPIG GV CN. üùõüý. 6JGUG RNCUOKFU JCXG DGGP GZVGPUKXGN[ UVWFKGF KP UGXGTCN NCDU þ)WPIG üùõø÷ 5VCTM GV CN. üùùö÷ 5EJCHHTCVJ GV CN. üùùù÷ /GKPJCTFV GV CN. üùù7ý. 6JG[ FKURNC[ C UVTWEVWTCN QTICPKUCVKQP CNUQ HQWPF KP QVJGT HWPICN NKPGCT RNCUOKFU CPF CTG TGOKPKUEGPV QH XKTWUÿNKMG IGPGVKE GNGOGPVU. 6JG K. lactis RNCUOKFU CTG UVCDN[ OCKPVCKPGF KP VJG E[VQRNCUO YJGTG VJG[ CTG TGRNKECVGF CPF VTCPUETKDGF D[ RNCUOKF GPEQFGF RQN[OGTCUGU TGNCVGF VQ OKÿ VQEJQPFTKCN RQN[OGTCUGU. #P ÉWRUVTGCO EQPUGPUWU UGSWGPEGÌ þð%5ý VJCV KU PQV TGEQIPKUGF D[ PWENGCT 40# RQN[OGTCUGU FGHKPGU VJG RTQOQVGTU QH VJG RNCUOKF 14(U þ4QOCPQU CPF $Q[F üùõõý. ©CEJ RNCUOKF EQPVCKPU C NCTIG 14( þ6CDNG üý VJCV GPEQFGU C &0#ÿRQN[OGTCUG þ&02ý CU C HWUKQP RTQVGKP YKVJ C RNCUOKFÿURGEKHKE VGTOKPCNÿGPFÿDKPFKPI RTQVGKP þ62ý VJCV UGTXGU CU TGRNKECVKQP RTKOGT. 6JG NCTIGT RNCUOKF þMû÷ R)-úûý EQPVCKPU PKPG CFFKVKQPCN 14(U GPEQFKPI OCKPVGPCPEG HWPEVKQPU KPENWFKPI C RTQVGKP TGSWKTGF HQT TGRNKECVKQP KPKVKCVKQP, C UKPINGÿUVTCPFGF &0# DKPFKPI RTQVGKP CPF C RWVCVKXG JGNKECUG. 6JG UOCNNGT Mü þR)-úüý RNCUOKFU ECTT[ QPN[ ò 14(U, VJTGG QH YJKEJ CTG
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
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TGNCVGF VQ VJG MKNNGT RJGPQV[RG. 14(U û CPF ò GPEQFG VJG MKNNGT VQZKP QT É\[OQEKPÌ, C VTKOGTKE UGETGVGF RTQVGKP. 6JG !ÿ CPF "ÿUWDWPKVU QH VJG \[OQEKP CTG RTQEGUUGF HTQO CP !"ÿRTGEWTUQT þMü 14(ûý QH YJKEJ VJG !ÿ UWDWPKV KU 0ÿIN[EQU[NCVGF. 6JG #ÿ UWDWPKV þÄÿVQZKPý GPEQFGF D[ 14(ò KU VJG E[VQVQZKE FGVGTOKPCPV þ$WVNGT GV CN. üùùüCýþUGG 5GEVKQP 3.òý. 14(3 KU CP KOOWPKV[ FGVGTOKPCPV QH WPMPQYP OQNGEWNCT PCVWTG. 9JGTGCU VJG NCTIGT Mû RNCUOKF KU UGNHÿUWUVCKPKPI, Mü TGSWKTGU Mû HWPEVKQPU HQT OCKPVGPCPEG þ5EJCHHTCVJ GV CN. üùùôý. (QT C OQTG FGVCKNGF FGUETKRVKQP QH VJG RNCUOKF CPF VJG K. lactis \[OQEKP VJG TGCFGT KU TGHGTTGF VQ C TGEGPV TGXKGY þ5EJCHHTCVJ CPF $TGWPKI ûöööý. 6CDNG 1. 2TQVGKP EQFKPI IGPGU QP K. lactis MKNNGT RNCUOKFU PCOG MüñR)-úü: 14(ü 14(û 14(3 14(ò MûñR)-úû 14(ü 14(û 14(3 14(ò 14(ô 14(ø
HWPEVKQP
14(7 14(õ 14(ù 14(üö
402! ! ! TGRNKECVKQP KPKVKCVKQP þ/E0GGN CPF 6COCPQK üùùü÷ 6QOOCUKPQ üùùüý
Müÿ62ñ&02 <[OQEKPþ½¾ý KOOWPKV[ \[OQEKP þÄý ! Mûÿ62ñ&02 O40# ECRRKPI JGNKECUG 55$ 402
4GHGTGPEG þ-KVCFC CPF )WPIG üùõõý þ5VCTM CPF $Q[F üùõøý þ6QMWPCIC GV CN. üùõ7ý þ$WVNGT GV CN. üùùüD÷6QMWPCIC GV CN. üùõùý þ6CMGFC GV CN. üùùø÷6QOOCUKPQ GV CN. üùõõý þúCTUGP GV CN. üùùõý þ5VCTM GV CN. üùùöý þ5EJCHHTCVJ CPF /GCEQEM ûööüý þ5EJCHHTCVJ GV CN. üùùô÷9KNUQP CPF /GCEQEM üùõõý þ5EJCHHTCVJ GV CN. üùù7ý
5KPEG VJG MKNNGT RNCUOKFU CTG TGOCTMCDN[ KPFGRGPFGPV QH JQUV HWPEVKQPU, VJG[ OC[ HWPEVKQP CU XGEVQTU YKVJ C DTQCF JQUV TCPIG. 2NCUOKF URGEKHKE RTQOQVGTU JCXG DGGP WUGF VQ FTKXG GZRTGUUKQP QH IGPGVKE OCTMGTU CPF TGEQODKPCPV IGPGU þ5EJKEMGN GV CN. üùùøý. 6JGUG IGPGU YGTG KPVTQFWEGF KPVQ VJG MüñMû IGPQOG D[ C TGEQODKPCÿ VKQPÿDCUGF E[VQRNCUOKE UJWHHNKPI U[UVGO þ5EJCHHTCVJ GV CN. üùùùý. *QYGXGT, FGURKVG VJG JKIJ EQR[ PWODGT þ$ ôöý RTQVGKP NGXGNU YGTG RQQT EQORCTGF VQ PWENGCT GRKÿ UQOCN RNCUOKFU. ©XGP YJGP 40# NGXGNU YGTG EQORCTCDNG VQ PWENGCT GPEQFGF VTCPUETKRVU, RTQVGKP NGXGNU YGTG OWEJ NQYGT KPFKECVKPI RQQT VTCPUNCVKQP QH VJGUG 40#U þ4GNKGPG CPF 5CUPCWUMCU üùù7ý. 6.2.2.3 Circular nuclear plasmids #P CFXCPVCIG QH K. lactis IGPGVKEU CPF DKQVGEJPQNQI[ KU VJG CXCKNCDKNKV[ QH OWNVKÿ EQR[ GRKUQOCN XGEVQTU. 6JKU IQGU DCEM VQ VJG RKQPGGTKPI YQTM QH (WMWJCTC CPF EQÿ YQTMGTU YJQ UETGGPGF OCP[ [GCUV URGEKGU HQT ET[RVKE GRKUQOCN RNCUOKFU. 6JG[ HQWPF C ü.ø±OÿRNCUOKF, R-&ü, KP Kluyveromyces lactis var. drosophilarum þ%JGP GV CN. üùõøý. 6JKU RNCUOKF ECP DG UVCDN[ OCKPVCKPGF KP K. lactis XCT. lactis CPF JCU
ü7ø
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
UKPEG DGGP FGXGNQRGF KPVQ C UGTKGU QH E.coli K. lactis QT E.coli K. lactis S. cerevisiae UJWVVNG XGEVQTU þ%JGP GV CN. üùõù÷ $KCPEJK GV CN. üùùü÷9oUQNQYUMKÿ úQWXGN GV CN. üùùøý. 6JG QTICPKUCVKQP QH R-&ü KU UKOKNCT VQ VJCV QH VJG û±Oÿ&0# QH S. cerevisiae: 6JG TGRNKECVKQP QTKIKP QXGTNCRU YKVJ QPG QH VJG VYQ KPXGTVGF TGÿ RGCVU CPF VJTGG IGPGU CPF C EKUÿCEVKPI UVCDKNKV[ NQEWU CTG KPXQNXGF KP TGRNKECVKQP, RCTVKVKQPKPI CPF EQR[ PWODGT EQPVTQN QH VJG GRKUQOG þ$KCPEJK GV CN. üùùüý. 6.2.3 /KVQEJQPFTKCN &0# 6JG UK\G QH VJG OKVQEJQPFTKCN &0# KU CDQWV 3ù MD þ9oUQNQYUMKÿúQWXGN GV CN. üùùøý CPF UGSWGPEGF OKVQEJQPFTKCN EQPVKIU EQORTKUG CDQWV û7 MD þ$QNQVKPÿ (WMWJCTC GV CN. ûöööý. #OQPI VJG UGSWGPEGF IGPGU CTG COX1 þE[VQEJTQOG QZKÿ FCUG UWDWPKV üý, COX2 þUWDWPKV ûý, COX3 þUWDWPKV3ý, ATP9 þUWDWPKV ù QH VJG OKVQÿ EJQPFTKCN #62CUGý, VAR1 þTKDQUQOCN RTQVGKPý, VJG ûü5 T40# IGPG, CPF UGXGTCN V40# IGPGU. +PVTQP UGSWGPEGU JCXG DGGP KFGPVKHKGF KP VJG ATP9 CPF ûü5 T40# IGPG UGSWGPEGU. # PQPCPWENGQVKFG ô'ÿþ#ñ6ý6#6##)6#ÿ3' EQPUGPUWU UGSWGPEG TGEQIPK\GF D[ VJG [GCUV OKVQEJQPFTKCN 40# RQN[OGTCUGU KU RTGUGPV KP HTQPV QH VJG VTCPUETKRVKQP WPKVU þ1UKPIC GV CN. üùõûý. /KVQEJQPFTKCN &0# COQWPVU WR VQ üö ó QH VQVCN EGNNWNCT &0#, UWIIGUVKPI CP CXGTCIG EQR[ PWODGT QH 3ö þ9oUQNQYUMKÿúQWXGN GV CN. üùùøý. 0WOGTQWU )%ÿTKEJ ENWUVGTU VJCV CTG TGEQODKPCVKQP JQV URQVU KP S. cerevisiae CTG RTGUGPV KP VJG K. lactis DWV PQV KP VJG K. marxianus OKVQEJQPFTKCN &0# þ4CIPKPK CPF (WMWJCTC üùõõý. &GURKVG VJG HCEV VJCV K. lactis KU RGVKVGÿPGICVKXG OKVQEJQPFTKCN &0# ECP TGEQODKPG CPF IKXG TKUG VQ OV RGVKVGU KP EGTVCKP OWVCPV DCEMITQWPFU þUGG 5GEVKQP 3.3ý 6.2.4 /CVKPI V[RGU K. lactis FKRQKFU CTG WPUVCDNG CPF URQTWNCVG GXGP QP TKEJ OGFKWO. 6JG RKQPGGTKPI YQTM QH *CNXQTUQP KP VJG GCTN[ UKZVKGU NCKF VJG ITQWPF HQT VJG IGPGVKEU QH K. lactis D[ GUVCDNKUJKPI IGPGVKECNN[ OCTMGF UVTCKPU YKVJ FGHKPGF OCVKPI V[RG þ*GTOCP CPF 4QOCP üùøø÷ 9oUQNQYUMKÿúQWXGN GV CN. üùùøý. +UQNCVKQP QH CP !ûÿNKMG IGPG þ;WCP GV CN. üùù3ý UWIIGUVGF VJCV OCVKPIÿV[RGU CTG EQPVTQNNGF KP C UKOKNCT YC[ CU KP S. cerevisiae. +PFGGF, UKOKNCTKV[ YCU EQPHKTOGF D[ VJG EJCTCEVGTK\CVKQP QH HML$, HMRa CPF MAT NQEK þ#UVTQO GV CN. ûöööý. HML CPF HMR NQEK CTG UKNGPEGF KP C 5KTû CPF 5KTò FGRGPFGPV OCPPGT þ#UVTQO CPF 4KPG üùùõý DWV VJGTG CTG C PWODGT QH FKHHGTGPEGU CU EQORCTGF VQ S. cerevisiae. +P VJG HML$ NQEWU, DGUKFGU VJG VYQ IGPGU !ü CPF !û TGUGODNKPI VJG S. cerevisiae IGPGU, C VJKTF NQPI 14(, !3 YCU HQWPF þ#UVTQO GV CN. ûöööý. 6JKU IGPG KU RCTV QH MAT$ DWV KU CNUQ GZRTGUUGF HTQO HML$ KP ! UVTCKPU. &GNGVKQP QH VJG IGPG TGFWEGF OCVKPI GHHKEKGPE[ DWV VJG RTGUÿ GPEG QH ½3 CV GKVJGT HML QT MAT YCU UWHHKEKGPV HQT OCVKPI. 6JG RWVCVKXG õù7 COKPQ CEKFU QH !3 FKF PQV TGXGCN CP[ UKOKNCTKV[ VQ IGPGU KP VJG FCVC DCUG. 6JG HMRC NQEWU EQPVCKPU VYQ IGPGU Cü CPF Cû CPF KPFKTGEV GXKFGPEG UWIIGUVU VJCV KP FKRQKF EGNNU CP Cüñ!û EQORNGZ GZKUVU VJCV TGRTGUUGU JCRNQKF URGEKHKE IGPGU NKMG KP S. cerevisiae.
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
ü77
#UVTQO GV CN. þ#UVTQO GV CN. ûöööý CNUQ UJQYGF VJCV UKNGPEKPI QH HML YCU FWG VQ VJG RTGUGPEG QH QPG EKUÿCEVKPI UKNGPEGT HNCPMGF D[ C UGSWGPEG EQPHGTTKPI #45 HWPEVKQP. +V YCU RQUUKDNG VQ FGNGVG VJG #45 YKVJQWV CHHGEVKPI UKNGPEKPI EQPHKTOKPI VJCV FGURKVG VJG GXQNWVKQPCT[ EQPUGTXCVKQP QH VJKU CTTCPIGOGPV #45 HWPEVKQP KU PQV CP GUUGPVKCN RTGTGSWKUKVG HQT UKNGPEKPI. 5KNGPEKPI YCU UJQYP VQ DG FGRGPFGPV QP K. lactis 5KTû CPF VQ C NGUUGT GZVGPV QP 5KTò. 6JG RTGUGPEG QH VJGUG HCEVQTU ETGCVGU C URGEKCNK\GF EJTQOCVKP UVTWEVWTG CV VJG UKNGPEGTU CU UJQYP D[ VJG TGUKUVCPEG VQ &0# OGVJ[NCVKQP. +P CFFKVKQP, K. lactis sir2 OWVCPVU CTG J[RGTÿUGPUKVKXG VQ ECVKQPU NKMG 0Kûï CPF %Wûï CPF GVJKFKWO DTQOKFG þ#UVTQO GV CN. ûöööý. 6JG TGNCVKQP QH VJKU RJGPQV[RG VQ VJG MPQYP 5KTû HWPEVKQP KU PQV WPFGTUVQQF. #P HO GPFQPWENGCUG IGPG JCU PQV DGGP KFGPVKHKGF UQ HCT. +H KV GZKUVU, KV OC[ DG GZRTGUUGF CV C XGT[ NQY TCVG KP UQOG UVTCKPU FKURNC[KPI CP CRRCTGPV UVCDNG OCVKPIÿ V[RG, UKPEG KV JCU DGGP QDUGTXGF VJCV UQOG QH VJGUG UVTCKPU ECP UYKVEJ YKVJ C NQY HTGSWGPE[ þ
6.3 þJ[UKQNQI[ 6.3.1 %CTDQP CPF GPGTI[ OGVCDQNKUO 6.3.1.1 Introduction +P Kluyveromyces lactis, CU KP OQUV QTICPKUOU, IN[EQN[UKU CPF VJG VTKECTDQZ[NKE CEKF E[ENG CTG VJG OCKP RCVJYC[U HQT VJG WVKNK\CVKQP QH UWICTU CPF VJG IGPGTCVKQP QH GPGTI[ þ#62ý. 5KOKNCTN[, INWEQUG KU VJG OQUV RTGHGTTGF UWICT, DWV Kluyveromyces URGEKGU ECP ITQY QP C YKFG XCTKGV[ QH UWICTU CPF QVJGT ECTDQP UQWTEGU. #RCTV HTQO NCEVQUG, UVTCKPU QH VJKU URGEKGU ECP ITQY HQT KPUVCPEG QP EGNNQDKQUG, OGNG\KVQUG, Z[NKVQN, OCPPKVQN, NCEVCVG, UWEEKPCVG, DWVCPGÿû,3ÿFKQN þYYY.EDU.MPCY.PNñ[GCUVý, CPF UGXGTCN COKPQ CEKFU þ
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-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
ü7ù
(KI. ÿ. ;GCUV ECTDQP CPF GPGTI[ OGVCDQNKUO. 6JG HKIWTG TGRTGUGPVU C J[RQVJGVKECN [GCUV EGNN YKVJ HGCVWTGU HTQO S. cerevisiae CPF QVJGT [GCUVU. 6JG VJKEM NKPG YKVJ DNCEM FQVU TGRTGÿ UGPVU VJG EQOOQP IN[EQN[VKE VTWPM CPF VJG 6%# E[ENG. &CUJGF NKPGU FGPQVG TGCEVKQPU VJCV CTG WUGF FWTKPI ITQYVJ KP PQPÿHGTOGPVCDNG ECTDQP UQWTEGU. úKPGU YKVJ FQVU CPF FCUJGU KPÿ FKECVG VJCV, FGRGPFKPI QP VJG [GCUV IGPGTC, UQOG QH VJG FKUCEEJCTKFGU TGRTGUGPVGF OC[ DG J[FTQN[\GF QWVUKFG VJG EGNN QT VTCPURQTVGF CU UWEJ KPVQ KV CPF J[FTQN[\GF KP VJG E[VQRNCUO. %KTENGU KP OGODTCPGU KPFKECVG VTCPURQTV U[UVGOU QT RTQVGKPU NQECVGF KP UWEJ UVTWEVWTGU. úKPGU IQKPI VJTQWIJ VJG OKVQEJQPFTKCN OGODTCPG FQ PQV PGEGUUCTKN[ KORNKECVG VJCV VJG EQTÿ TGURQPFKPI OGVCDQNKVG HTGGN[ ETQUUGU KV. 6JG OCNKE GP\[OG KU PQV TGRTGUGPVGF. 0Q QTICPGNNGU DGUKFG VJG OKVQEJQPFTKQP CTG FTCYP. 6JG HQNNQYKPI PQPÿUVCPFCTF CDDTGXKCVKQPU KP RCVJYC[U FKHHGTGPV HTQO IN[EQN[UKU JCXG DGGP WUGF: ø2), øÿRJQURJQINWEQPCVG: 4WNô2, TKDWNQUG ôÿ RJQURJCVG: 4ôR, TKDQUG ôÿ RJQURJCVG÷ îWNô2, Z[NWNQUG ôÿRJQURJCVG÷ 572, UGFQJGRVWNQUG 7ÿ RJQURJCVG÷ ©ò2, GT[VJTQUG òÿRJQURJCVG÷)32, IN[EGTQN 3ÿRJQURJCVG÷ 2K, 'KPQTICPKE" RJQUÿ RJCVG÷ 1##, QZCNQCEGVCVG÷ í, WDKSWKPQPG÷ DEü, E[VQEJTQOGU÷ %[V QZ, E[VQEJTQOG QZKFCUG. 4GRTKPVGF HTQO (NQTGU GV CN. þûöööý YKVJ RGTOKUUKQP HTQO ©NUGXKGT.
KVU QRVKOWO ITQYVJ VGORGTCVWTG QH 33·%, VJG IGPGTCVKQP VKOG OKIJV GXGP DG TGÿ FWEGF VQ 7ô OKP þ5VGGPUOC GV CN. üùõõý. /QUV TGUWNVU FKUEWUUGF DGNQY TGHGT VQ VJKU UVTCKP CPF KVU FGTKXCVKXGU, WPNGUU UVCVGF QVJGTYKUG. /CP[ IGPGU KPXQNXGF KP INWEQUG OGVCDQNKUO JCXG DGGP HQWPF KP C UETGGP HQT OWVCPVU VJCV DGECOG UGPUKVKXG VQ VJG TGURKTCVKQP KPJKDKVQT CPVKO[EKP # KP VJG RTGUÿ GPEG QH JKIJ INWEQUG EQPEGPVTCVKQPU, VJG 4CIÿ RJGPQV[RG. 5GXGTCN PCVWTCN K. lactis KUQNCVGU CTG UGPUKVKXG VQ VJKU FTWI, YJGTGCU QVJGTU CTG TGUKUVCPV þ(GTTGTQ GV CN. üùõüý. 6JG TGUKUVCPV UVTCKPU RTQDCDN[ IGPGTCVG UWHHKEKGPV GPGTI[ HTQO HGTOGPVCVKQP VQ UWUÿ VCKP ITQYVJ. /WVCVKQPU KP IGPGU KPXQNXGF KP IN[EQN[UKU CPF CNEQJQNKE HGTOGPVCVKQP YKNN FKUCDNG VJKU ECRCEKV[. #V VJG NCVGUV EQWPV, QXGT VYGPV[ 4CIÿ EQORNGOGPVCVKQP ITQWRU JCXG DGGP HQWPF þ/. 9oUQNQYUMKÿúQWXGN , RGTUQPCN EQOOWPKECVKQPý CPF VJGUG KPENWFG IGPGU GPEQFKPI UVTWEVWTCN CU YGNN CU TGIWNCVQT[ RTQVGKPU KPXQNXGF KP IN[EQN[UKU CPF HGTOGPVCVKQP. 5GXGTCN QH VJGUG IGPGU YKNN RCUU D[ KP UWDUGSWGPV UGEÿ VKQPU QH VJKU EJCRVGT. 6.3.1.2 Glucose uptake +P EQPVTCUV VQ S.cerevisiae, KP YJKEJ C NCTIG HCOKN[ QH JGZQUG VTCPURQTVGTU KU RTGÿ UGPV, QPN[ HQWT IGPGU HQT VJG WRVCMG QH JGZQUGU JCXG DGGP HQWPF UQ HCT KP K. lactis. RAG1 GPEQFGU C WPKSWG QT VJG OCKP NQYÿCHHKPKV[ INWEQUG VTCPURQTVGT KP UQOG UVTCKPU þ2TKQT GV CN. üùù3C÷ 9oUQNQYUMKÿúQWXGN GV CN. üùùûC÷ 9oUQNQYUMKÿúQWXGN GV CN. üùùûD÷ )QHHTKPK GV CN. üùùöý, YJGTGCU KP QVJGT UVTCKPU VJKU NQEWU ECTKGU VYQ VCPFGON[ CTTCPIGF JGZQUG VTCPURQTVGT IGPGU, KHT1 CPF KHT2, YKVJ õòó UGÿ SWGPEG KFGPVKV[ GCEJ QH YJKEJ EQPHGTU C 4CIï RJGPQV[RG þ9GKTKEJ GV CN. üùù7ý. #NN VJTGG VTCPURQTVGT IGPGU JCXG JKIJ UGSWGPEG UKOKNCTKV[ CPF KV CRRGCTU VJCV RAG1 JCU CTKUGP D[ C FGNGVKQP GXGPV DGVYGGP VJG VYQ KHT IGPGU UWEJ VJCV RAG1 KU KFGPVKECN VQ KHT1 GZEGRV HQT VJG NCUV VJTGG COKPQ CEKFU, YJKEJ QTKIKPCVG HTQO KHT2. (TWEÿ VQUG CPF ICNCEVQUG CTG CNUQ VCMGP WR D[ VJGUG INWEQUG VTCPURQTVGTU þ9oUQNQYUMKÿ úQWXGN GV CN. üùùûC÷ 9GKTKEJ GV CN. üùù7ý. 5VTCKP %$5 û3øö KU C PCVWTCN 4CIÿ KUQNCVG
üõö
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
KP YJKEJ ITQYVJ QP INWEQUG KU UVTKEVN[ FGRGPFGPV QP TGURKTCVKQP FWG VQ C RQKPVÿ OWVCVKQP KP VJG RAG1 IGPG þ)QHHTKPK GV CN. üùõù÷)QHHTKPK GV CN. üùùöý. +P CFFKVKQP VQ VJG NQY CHHKPKV[ INWEQUG VTCPURQTVGTU, C JKIJ CHHKPKV[ VTCPURQTVGT GPEQFGF D[ VJG HGT1 IGPG KU RTGUGPV KP OCP[ UVTCKPU þ$KNNCTF GV CN. üùùøý. +P EQPÿ VTCUV VQ RAG1 CPF KHT1, YJKEJ JCXG VJG UCOG WRUVTGCO UGSWGPEGU CPF CTG KPÿ FWEGF D[ HGTOGPVCDNG ECTDQP UQWTEGU þ9oUQNQYUMKÿúQWXGN GV CN. üùùû÷ %JGP GV CN. üùùûD÷
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üõü
TWRVGF ECP ITQY QP INWEQUG, YJGTGCU VJG EQTTGURQPFKPI S. cerevisiae OWVCPVU ECP PQV þ*GKPKUEJ GV CN. üùù3ý. 6JGUG FCVC UWIIGUV VJCV KP K. lactis VJG RGPVQUG RJQUÿ RJCVG RCVJYC[ ECP D[RCUU VJGUG VYQ UVGRU QH IN[EQN[UKU. &KUTWRVKQP QH VJG IGPG HQT VTCPUCNFQNCUG þKlTAL1ý KP VJG Klpfk1 Klpfk2 FQWDNG OWVCPV KPFGGF CDQNKUJGU ITQYVJ þìCEQD[ GV CN. üùù3ý. 6TCPUCNFQNCUG VQIGVJGT YKVJ VTCPUMGVQNCUG GPEQFGF D[ VJG KlTKL1 IGPG þìCEQD[ CPF *GKPKUEJ üùù7ý EQPXGTVU TKDWNQUGÿôÿRJQURJCVG KPVQ HTWEVQUGÿøÿRJQURJCVG CPF 3ÿRJQURJQIN[EGTCNFGJ[FG. 6JG HKPFKPI VJCV OWVCVKQPU KP VJG RJQURJQIN[EGTCVG IGPG, KlPGK1 þ(QWTPKGT GV CN. üùùöý, HTQO VJG NQYGT RCTV QH IN[EQN[UKU, DNQEM ITQYVJ QP INWEQUG þ(NGGT, SWQVGF KP ìCEQD[ GV CN. üùù3ý UWRRQTVU VJG KFGC VJCV KP K. lactis VJG RGPVQUG RJQUÿ RJCVG RCVJYC[ ECP D[RCUU VJG HKTUV VYQ EQOOKVVGF UVGRU QH IN[EQN[UKU. 6JKU OGCPU VJCV, KP EQPVTCUV VQ S.cerevisiae, K. lactis JCU DQVJ C UWHHKEKGPVN[ CEVKXG RGPVQUG RJQURJCVG RCVJYC[ CPF VJG ECRCEKV[ VQ TGÿQZKFK\G VJG 0#&2* HQTOGF þ)QP\hNG\ 5KUQ GV CN. üùùøý. 1Z[IGP WRVCMG GZRGTKOGPVU EQPHKTOGF VJG RTGUGPEG QH C OKVQÿ EJQPFTKCN 0#&2* FGJ[FTQIGPCUG KP K. lactis, CP GP\[OG PQV HQWPF KP S.cerevisiae. 6JKU GP\[OG KU QTKGPVGF VQ VJG GZVGTPCN HCEG QH VJG KPPGT OKVQEJQPÿ FTKCN OGODTCPG CPF ECP EQWRNG VJG QZKFCVKQP QH E[VQUQNKE 0#&2* VQ VJG TGURKTCÿ VQT[ EJCKP þ1XGTMCOR GV CN. ûööûý. 6JG KPCDKNKV[ QH VJG rag2 OWVCPV VQ ITQY KP VJG RTGUGPEG QH CPVKO[EKP # þ)QHHTKPK GV CN. üùõùý KU PQV ECWUGF D[ CP KPCDKNKV[ VQ HQTO GVJCPQN CU RTGXKQWUN[ TGRQTVGF þ)QHHTKPK GV CN. üùùüý, DWV D[ C TGFQZ FGHGEV. 6JG 0#&2* HQTOGF KP VJG RGPVQUG RJQURJCVG RCVJYC[ ECP PQV DG TGÿQZKFK\GF YJGP TGURKTCVKQP KU DNQEMGF þ)QP\hNG\ 5KUQ GV CN. üùùøý. 6JG KPVGTEQPXGTUKQP QH FKJ[FTQZ[CEGVQPG RJQURJCVG CPF IN[EGTCNFGJ[FGÿ3ÿ RJQURJCVG KU ECVCN[UGF D[ VTKQUG RJQURJCVG KUQOGTCUG GPEQFGF D[ C UKPING IGPG, TPI1. +P EQPVTCUV VQ C UKOKNCT OWVCPV KP S.cerevisiae, C K. lactis tpi1 OWVCPV ECP UVKNN ITQY QP INWEQUG þ%QORCIPQ GV CN. üùùùý. 6JKU FKHHGTGPEG OKIJV CICKP DG ECWUGF D[ C OQTG RTQOKPGPV TQNG QH VJG RGPVQUG RJQURJCVG RCVJYC[ KP K. lactis, UKPEG INWEQUG OGVCDQNK\GF XKC VJKU RCVJYC[ YKNN HQTO QPN[ VYQ VJKTFU VJG COQWPV QH FKJ[FTQZ[CEGVQPG RJQURJCVG KP EQORCTKUQP VQ VJG FKTGEV RCVJYC[ XKC INWEQUG KUQOGTCUG CPF RJQURJQHTWEVQMKPCUG. 6.3.1.4 Pyruvate metabolism 6JG EQPXGTUKQP QH IN[EGTCNFGJ[FGÿ3ÿRJQURJCVG VQ R[TWXCVG QEEWTU KP C PWODGT QH UVGRU HQT YJKEJ FKHHGTGPEGU YKVJ QVJGT [GCUVU JCXG PQV DGGP TGRQTVGF. 2[TWXCVG, JQYGXGT, KU CP KORQTVCPV DTCPEJRQKPV KP OGVCDQNKUO. ðPFGT QZ[IGPÿNKOKVGF EQPFKÿ VKQPU 0#&* ECPPQV DG þEQORNGVGN[ý TGQZKFK\GF CPF CNEQJQNKE HGTOGPVCVKQP UVCTVU YKVJ VJG FGECTDQZ[NCVKQP QH R[TWXCVG VQ CEGVCNFGJ[FG, YJKEJ KU HWTVJGT TGFWEGF VQ GVJCPQN, UKOWNVCPGQWUN[ þTGýIGPGTCVKPI 0#&ï HTQO 0#&* þ(KI. 3ý. +PCEVKXCVKQP QH VJG R[TWXCVG FGJ[FTQIGPCUG EQORNGZ D[ FKUTWRVKQP QH VJG PDA1 IGPG GPEQFKPI VJG FGECTDQZ[NCVKPI ©ü½ UWDWPKV NGCFU VQ UGXGTG ITQYVJ FGHGEVU KP K. lactis, CNVJQWIJ ITQYVJ KU PQV EQORNGVGN[ DNQEMGF þ
üõû
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
0#&* R[TWXCVG
ü
CEGVCNFGJ[FG
0#& + û
GVJCPQN
0#&þ2) + 3
0#&þ2)*
CEGVCVG (KI. 3. (GTOGPVCVKQP. ü. R[TWXCVG FGECTDQZ[NCUG÷ û. CNEQJQN FGJ[FTQIGPCUG÷ 3. CNFGJ[FG FGJ[FTQIGPCUG.
YJGP DTQMGP FQYP. +PVGTGUVKPIN[, GVJCPQN HQTOCVKQP YCU GNGXCVGF KP VJG pda1 OWÿ VCPV. 6JKU UWIIGUVU VJGTGHQTG, VJCV KV KU VJG CEVKXKV[ QH VJG R[TWXCVG FGJ[FTQIGPCUG EQORNGZ, URGEKHKECNN[ KVU GHHKEKGPV EQORGVKVKQP YKVJ R[TWXCVG FGECTDQZ[NCUG HQT R[ÿ TWXCVG, YJKEJ TGPFGTU K. lactis C %TCDVTGG PGICVKXG [GCUV þUGG 5GEVKQP 3.ü.ôý. 6JG TGOCKPKPI ITQYVJ QH VJG pda1 OWVCPV KU ECWUGF D[ VJG UQÿECNNGF 2&* D[ÿ RCUU, YJKEJ HQTOU #E%Q# D[ VJG UGSWGPVKCN CEVKXKV[ QH R[TWXCVG FGECTDQZ[NCUG þ2FEý, CEGVCNFGJ[FG FGJ[FTQIGPCUG þ#NFý, CPF CEGV[NÿEQGP\[OG # U[PVJCUG þ#EUý. 5KPEG CV NGCUV 2FE CPF #EU CTG NQECVGF KP VJG E[VQUQN, VJG #E%Q# HQTOGF KU E[VQUQÿ NKE. +V CRRGCTU VJCV VTCPURQTV KPVQ OKVQEJQPFTKC KU NKOKVKPI UKPEG CFFKVKQP QH UOCNN COQWPVU QH úÿECTPKVKPG CNOQUV EQORNGVGN[ TGUVQTG ITQYVJ þ
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üõ3
QZ[IGP EQPEGPVTCVKQPU, C EQPFKVKQP TCRKFN[ QEEWTTKPI KP UJCMG HNCUM EWNVWTGU þ-KGTU GV CN. üùùõý. +P CGTQDKE UJCMG HNCUM EWNVWTGU ITQYP CV JKIJ INWEQUG EQPEGPVTCVKQPU, HQTOCVKQP QH GVJCPQN QPN[ UGVU KP CV JKIJ EGNNWNCT FGPUKVKGU YJGP QZ[IGP UWRRN[ DGEQOGU NKOKVKPI. +P EJGOQUVCV EWNVWTGU, GVJCPQN HQTOCVKQP YCU DGNQY VJG FGVGEÿ VKQP NKOKV KP C DTQCF TCPIG QH FKNWVKQP TCVGU þ-KGTU GV CN. üùùõ÷
üõò
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
VQUG RJQURJCVG RCVJYC[ þ$Q\\K GV CN. üùù7÷ 1XGTMCOR GV CN. ûööûý OGPVKQPGF RTGÿ XKQWUN[ KP VJKU EJCRVGT. 6JG RTGUGPEG QH HQWT CNEQJQN FGJ[FTQIGPCUGU KP CP QTICPKUO YKVJ C RTGFQOKÿ PCPVN[ TGURKTCVQT[ OGVCDQNKUO OKIJV TGHNGEV VJG PGEGUUKV[ VQ EQPVTQN KPVTCEGNNWNCT NGXGNU QH VJG JKIJN[ VQZKE CEGVCNFGJ[FG. ðPFGT NQY QZ[IGP EQPFKVKQPU, HGTOGPVCÿ VKQP UGVU KP VQ TGNKGXG VJG TGFQZ RTQDNGO D[ TGQZKFK\KPI 0#&* þ(KI. 3ý. 1PEG VJG GP\[OG R[TWXCVG FGECTDQZ[NCUG KU CEVKXG, UWHHKEKGPV CNEQJQN FGJ[FTQIGPCUG ECRCEÿ KV[ UJQWNF DG RTGUGPV VQ RTGXGPV VJG DWKNF WR QH VQQ JKIJ CEGVCNFGJ[FG EQPEGPVTCÿ VKQPU. /QTGQXGT, VJKU KFGC CNUQ TGSWKTGU VJCV CEGVCNFGJ[FG FGJ[FTQIGPCUG YJKEJ QZKFK\GU CEGVCNFGJ[FG VQ CEGVCVG KU MGRV WPFGT EQPVTQN, UKPEG VJKU TGCEVKQP YKNN GPÿ ITCXG VJG TGFQZ RTQDNGO D[ RTQFWEKPI 0#&þ2ý*. úKVVNG KU MPQYP CDQWV VJG þCEGVýCNFGJ[FG FGJ[FTQIGPCUGU KP K. lactis. 6.3.1.6 Acetate and ethanol consumption K. lactis ECP ITQY QP %ûÿECTDQP UQWTEGU. &WTKPI ITQYVJ QP GVJCPQN, CNN HQWT CNEQÿ JQN FGJ[FTQIGPCUGU CTG VTCPUETKDGF KP ECTDQPÿNKOKVGF EQPVKPWQWU EWNVWTGU þ
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üõô
(QT ITQYVJ QP GVJCPQN QT CEGVCVG, QZCNQCEGVCVG OWUV DG U[PVJGUK\GF VQ TGRNGPKUJ VJG 6%# E[ENG. 6JKU QEEWTU XKC VJG IN[QZ[NCVG E[ENG XKC VYQ URGEKHKE GP\[OGU, KUQEKVTCVG N[CUG CPF OCNCVG U[PVJCUG. /QTGQXGT, INWEQUGÿRJQURJCVGU CPF ð&2ÿ INWEQUG CTG TGSWKTGF CU DWKNFKPI DNQEMU KP EGNN YCNN DKQIGPGUKU, UVQTCIG QH ECTDQÿ J[FTCVGU, CPF IN[EQU[NCVKQP. 6JGUG OGVCDQNKVGU JCXG VQ DG U[PVJGUKUGF D[ INWÿ EQPGQIGPGUKU, YJKEJ TGSWKTGU VYQ URGEKHKE GP\[OGU VJCV ECVCN[UG VJG TGXGTUG TGCEÿ VKQPU QH VJG PQPÿTGXGTUKDNG UVGRU QH IN[EQN[UKU, HTWEVQUGÿDKUÿRJQURJCVCUG þ($2CUGý, CPF RJQURJQÿGPQNÿECTDQZ[MKPCUG þ2%-ý. úKVVNG KU MPQYP CDQWV VJG UVTWEVWTCN K. lactis IGPGU KPXQNXGF KP VJG IN[QZ[NCVG E[ENG CPF INWEQPGQIGPGUKU. 5QOG QH VJG TGNGXCPV IGPGU JCXG DGGP KUQNCVGF þ
üõø
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
KPVCEV 4CIõ. +P C rag8 OWVCPV, VJG NGXGN QH SCK1 O40# KU QPN[ òöó QH VJCV KP C YKNF V[RG UVTCKP þúGOCKTG GV CN. ûööûý. +P CFFKVKQP VQ FKTGEV VTCPUETKRVKQP TGIWNCVKQP D[ VJG RAG4 CPF RAG8 RTQFWEVU, INWEQUG OGVCDQNKUO OWUV DG KPVCEV HQT HWNN KPFWEVKQP QH VJG RAG1 IGPG. /WVCVKQPU KP VJG IGPGU HQT JGZQMKPCUG þRAG5ý CPF VJG INWEQUG VTCPURQTVGTU VJGOUGNXGU þRAG1/KHT1/KHT2ý CDQNKUJ VTCPUETKRVKQP HTQO VJG RAG1 QT KHT1 RTQOQVGTU þ%JGP GV CN. üùùû÷ 2TKQT GV CN. üùù3D÷ $CTKPCIC, üùùò÷ $TGWPKI GV CN. ûööö÷ /KNMQYUMK GV CN. ûööüý. #U GZRTGUUKQP QH VJG VTCPURQTVGT IGPGU VJGOUGNXGU KU UWDÿ LGEV VQ INWEQUG KPFWEVKQP CPFñQT INWEQUG TGRTGUUKQP, C EQPVTQN EKTEWKV KU ETGCVGF VJCV CNNQYU VJG EGNNU VQ CFLWUV INQDCN INWEQUG OGVCDQNKUO VQ VJG IN[EQN[VKE HNWZ CPF vice versa. *GPEG INWEQUG WRVCMG KU CWVQTGIWNCVGF CPF KP UVTCKPU YKVJ DQVJ KHT1 CPF KHT2 RTGUGPV, VJGUG IGPGU OWVWCNN[ EQPVTQN GCEJ QVJGT UWEJ VJCV VJG INWEQUG EQPÿ EGPVTCVKQP FGVGTOKPGU VJG GZRTGUUKQP NGXGN QH GCEJ QH VJG IGPGU þ/KNMQYUMK GV CN. ûööüý. 6JG FKHHGTGPEG YKVJ S. cerevisiae CRRGCTU VQ DG C TGUVTKEVKQP QH VJG IN[EQÿ N[VKE HNWZ WPFGT EQPFKVKQPU QH UWHHKEKGPV QZ[IGP UWRRN[ GXGP YJGP INWEQUG KU RTGÿ UGPV CV JKIJ EQPEGPVTCVKQPU KP VJG OGFKWO. 6JG EQPVTQN QH JGZQUG VTCPURQTVGT IGPG TGIWNCVKQP D[ QZ[IGP JCU PQV DGGP UVWFKGF, DWV OKIJV DG QH KORQTVCPEG KP VJKU TGÿ ICTF. )ETü, )ETû, CPF 4CRü CTG PGEGUUCT[ HQT HWNN KPFWEVKQP QH IN[EQN[UKU KP S. cerevisiae þðGOWTC CPF ìKICOK üùùû÷
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üõ7
CPF GAL83 YGTG KFGPVKHKGF. 6JG IGPGU FOG2 þKlSNF1ý CPF FOG1 þKlGAL83ý YGTG KUQNCVGF D[ VJGKT CDKNKV[ VQ CHHGEV fGTOGPVCVKXG CPF oZKFCVKXG gTQYVJ þ)QHHTKPK GV CN. üùùøý. 6JG OWVCPV RJGPQV[RG QH fog1 CPF VJG NCEM QH ETQUUÿJ[DTKFKUCVKQP YKVJ C FOG1 RTQDG KPFKECVG VJCV QPN[ QPG QH VJG UECHHQNF RTQVGKPU KU RTGUGPV KP K. lactis þ)QHHTKPK GV CN. üùùøý. 5KPEG KP VJGUG S. cerevisiae RTQVGKPU CRRGCT VQ EQPHGT UWDUVTCVG URGEKHKEKV[ VQ VJG 5PHü EQORNGZ CPFñQT CTG KPXQNXGF KP VCTIGVKPI VJG MKPCUG VQ URGEKHKE EQORCTVOGPVU, KV YKNN DG KPVGTGUVKPI VQ UGG JQY UKIPCNNKPI URGEKÿ HKEKV[ KU CUUWTGF KP K. lactis. $[ UWRRTGUUKQP QH C K. lactis fog2/Klsnf1 OWVCPV, VJG KlCAT8 IGPG JCU DGGP ENQPGF. +V EQWNF EQORNGOGPV C cat8 OWVCPV KP UKPING EQR[ þ)GQTKU GV CN. ûöööý. KLSIP4 YCU QDVCKPGF CU C OWNVKÿEQR[ UWRRTGUUQT QH C Klcat8 OWVCVKQP þì.ì. -TKLÿ IGT, 2J& VJGUKU *CNNGý. 5WTRTKUKPIN[, FGNGVKQP QH Klcat8 FQGU PQV CHHGEV GZRTGUUKQP QH VJG KlFBP1 CPF KlPCK1 IGPGU CPF ITQYVJ QP IN[EGTQN KU WPKORCKTGF þ)GQTKU GV CN. ûöööý. 6JWU, KP EQPVTCUV VQ S. cerevisiae INWEQPGQIGPGUKU KP K. lactis, KV KU PQV FGRGPFGPV QP -N%CVõ þì.ì. -TKLIGT, 2J& VJGUKU *CNNGý. *QYGXGT, -N%CVõ KU TGSWKTGF HQT ITQYVJ QP CEGVCVG, GVJCPQN, CPF NCEVCVG UKPEG VTCPUETKRVKQP QH DQVJ KlACS IGPGU þúQFK GV CN. ûööüý, VJG IN[QZ[NCVG E[ENG IGPGU KlICL1 þ)GQTKU GV CN. ûöööý CPF KlMLS1 þì.ì. -TKLIGT, 2J& VJGUKU *CNNGý, CPF VJG NCEVCVG VTCPURQTVGT IGPG KlJEN1 þúQFK GV CN. ûööüý CTG TGFWEGF KP C Klcat8 OWVCPV. #PQVJGT KORQTVCPV TGIWNCVQT[ OGEJCPKUO KU VJG *CRûñ3ñòñô EQORNGZ. +P S.cerevisiae, VJKU EQORNGZ CEVKXCVGU IGPGU GPEQFKPI EQORQPGPVU QH VJG TGURKTCVQT[ EJCKP CPF VJG 6%# E[ENG KP VJG RTGUGPEG QH PQPÿHGTOGPVCDNG ECTDQP UQWTEGU. *Qÿ OQNQIU QH HAP2, HAP3, CPF HAP4 JCXG DGGP KUQNCVGF HTQO K. lactis D[ JGVGTQNQÿ IQWU EQORNGOGPVCVKQP QH VJG EQTTGURQPFKPI PWNN OWVCPVU QH S. cerevisiae þ/WNFGT GV CN. üùùò÷ 0IW[GP GV CN. üùùô÷ $QWTICTGN GV CN. üùùùý. *QYGXGT, FKUTWRVKQP QH VJG K. lactis IGPGU FQGU PQV NGCF VQ VJG KPCDKNKV[ VQ ITQY QP IN[EGTQN, NCEVCVG, GVJCPQN, QT CEGVCVG KP EQORNGZ OGFKC, YJGTGCU KP OKPKOCN OGFKC Klhap2 CPF Klhap4 OWÿ VCPVU ITGY NGUU VJCP VJG YKNF V[RG UVTCKP þúQFK GV CN. üùùõ÷ $QWTICTGN GV CN. üùùùý. 6JGUG FCVC UWIIGUV VJCV KP K. lactis TGURKTCVKQP FQGU PQV TGSWKTG VJG *CRûñ3ñòñô EQORNGZ. +PFGGF, VTCPUETKRVKQP QH KlCYC1, GPEQFKPI E[VQEJTQOG c, KU PQV CHHGEVGF KP C Klhap3 OWVCPV, FGURKVG VJG RTGUGPEG QH C DKPFKPI UKVG HQT VJG *CR EQORNGZ KP VJG KlCYC1 RTQOQVGT þ(TGKTGÿ2KEQU GV CN. üùù3÷ /WNFGT GV CN. üùùò÷ (TGKTGÿ2KEQU GV CN. üùùô÷ 4COKN GV CN. üùùõý. 6JG TGURKTCVQT[ IGPGU KlCYC1, KlQCR7, CPF KlQCR8, VJG NCVVGT VYQ GPEQFG UWDWPKVU QH VJG bcü EQORNGZ, CTG PQV TGRTGUUGF D[ INWEQUG KP VJG RTGUGPEG QH C PQPÿHGTOGPVCDNG ECTDQP UQWTEG þ/WNFGT GV CN. üùùôCý CNVJQWIJ KlCYC1 O40# KU CDUGPV HTQO EGNNU ITQYP KP ô ó INWEQUG þ/WNFGT GV CN. üùùòý CPF ECTDQP ECVCDQNKVG, TGRTGUUKQP QH VJG IGPG JCU DGGP TGRQTVGF HQT CPQVJGT UVTCKP þ(TGKTGÿ2KEQU GV CN. üùùôý. Respiration and fermentation. /KETQCTTC[ FCVC JCXG UJQYP VJCV KP S. cerevisiae VTCPUETKRVKQP QH QXGT 3öö IGPGU KU CHHGEVGF D[ VJG RTGUGPEG QT CDUGPEG QH OQNGEWNCT QZ[IGP þ6GT úKPFG GV CN. üùùùý. 4GRTGUUKQP CPF CEVKXCVKQP QH UQÿECNNGF J[RQZKE QT CPCGTQDKE IGPGU KU OGFKCVGF D[ JGOG, VJG 5UPøÿ6WRü EQORNGZ CPF UGXGTCN TGIWNCÿ VQTU, KPENWFKPI HAP1, ROX1, UPC2, MOX1 CPF MOX2 þ6GT úKPFG CPF 5VGGPUOC ûöö3ý. *QOQNQIU QH VJGUG IGPGU JCXG PQV DGGP KFGPVKHKGF UQ HCT KP K. lactis. #Nÿ
üõõ
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
VJQWIJ K. lactis ECPPQV UWUVCKP ITQYVJ KP VJG EQORNGVG CDUGPEG QH OQNGEWNCT QZ[ÿ IGP, KV TGCEVU VQ NQY QZ[IGP EQPEGPVTCVKQPU. KlHEM1 CPF KlHEM13 GPEQFKPI VJG HKTUV CPF UKZVJ UVGR KP JGOG DKQU[PVJGUKU, TGURGEVKXGN[, KPETGCUG VTCPUETKRVKQP WPFGT J[RQZKE EQPFKVKQPU. +PVGTGUVKPIN[, KlHEM13 þ)QP\hNG\ÿ&QOKPIWG\ GV CN. ûöööý CPF VQ C NQYGT GZVGPV KlHEM1 þ)QP\hNG\ÿ&QOKPIWG\ GV CN. üùù7ý CTG TGRTGUUGF D[ JGOG, VJWU ETGCVKPI CP CWVQTGIWNCVQT[ NQQR. KlCYC1 CPF VJG E[VQEJTQOG cü IGPG KlCYT1 þ)DGNUMC GV CN. üùùøý, CTG VKIJVN[ EQPVTQNNGF D[ QZ[IGP CPF JGOG, YJKNG VYQ QVJGT TGURKTCVQT[ IGPGU, KlQCR7 CPF KlQCR8, CTG QPN[ OQFGTCVGN[ JKIJGT GZRTGUUGF KP JGOGÿUWHHKEKGPV QT PQTOQZKE EQPFKVKQPU þ)QP\hNG\ÿ&QOKPIWG\ GV CN. ûöööý. #V NQYÿQZ[IGP EQPEGPVTCVKQPU, HGTOGPVCVKQP UGVU KP, DWV JQY VJKU KU TGIWNCVGF KU PQV ENGCT. 6JG HKTUV UVGR KP VJG HGTOGPVCVKQPÿURGEKHKE RCVJYC[ KU RGTHQTOGF D[ R[ÿ TWXCVG FGECTDQZ[NCUG, YJKEJ KP K. lactis KU GPEQFGF D[ C UKPING IGPG þ$KCPEJK GV CN. üùùøý. #U FGUETKDGF RTGXKQWUN[ þUGG 3.ü.ôý VTCPUETKRVKQP TGIWNCVKQP QH VJG IGPG KU EQORNGZ. +V KU CWVQTGIWNCVGF, KPFWEGF D[ INWEQUG CPF NQY QZ[IGP EQPEGPVTCVKQPU CPF TGRTGUUGF D[ GVJCPQN. )NWEQUG KPFWEVKQP QH VJG IGPG TGSWKTGU KPVCEV IN[EQN[UKU UKPEG OWVCVKQPU KP INWEQUG WRVCMG þRAG1ý, JGZQMKPCUG þRAG5ý, CPF INWEQUG KUQOÿ GTCUG þRAG2ý CU YGNN CU KP TGIWNCVQTU QH VJGUG IGPGU þRAG8, KlGGS1, SCK1ý CDQNÿ KUJ QT TGFWEG VTCPUETKRVKQP KP INWEQUG OGFKC þúW[VGP GV CN. üùù3÷ $KCPEJK GV CN. üùùø÷ úGOCKTG GV CN. ûööûý.. # PWNN OWVCVKQP KP RAG3, VJG JQOQNQI QH VJG PDC2 IGPG þ2TKQT GV CN. üùùøý UVTQPIN[ TGFWEGU VTCPUETKRVKQP QH KlPDC1, CPF KlTHI4 CPF KlTHI5, VYQ IGPGU KPÿ XQNXGF KP VJKCOKPG DKQU[PVJGUKU 6JKCOKPGR[TQRJQURJCVG þ622ý KU C EQHCEVQT QH UGXGTCN GP\[OGU, KPENWFKPI R[TWXCVG FGECTDQZ[NCUG CPF VJG ©ü! UWDWPKV QH VJG R[ÿ TWXCVG FGJ[FTQIGPCUG EQORNGZ. 6TCPUETKRVKQP QH KlPDA1, GPEQFKPI VJG NCVVGT, KU RCTVN[ TGRTGUUGF D[ 4CI3 þ6K\\CPK GV CN. üùùõý. 6JGUG FCVC RQKPV VQ C TQNG QH 4CI3 KP VJG QPUGV QH HGTOGPVCVKQP D[ CEVKXCVKPI R[TWXCVG FGECTDQZ[NCUG CPF TGFWEKPI R[TWXCVG FGJ[FTQIGPCUG CEVKXKV[ þ6K\\CPK GV CN. üùùõý. 6.3.2 LCEVQUG WVKNK\CVKQP 6JG CDKNKV[ VQ WVKNKUG NCEVQUG CU C ECTDQP UQWTEG KU TCTG COQPI [GCUVU. 6JG IGPGVKEU CPF RJ[UKQNQI[ QH NCEVQUG OGVCDQNKUO KP K. lactis JCU VJGTGHQTG DGGP UVWFKGF GZÿ VGPUKXGN[. &GITCFCVKQP QH NCEVQUG KP OKNM HQT NCEVQUGÿKPVQNGTCPV JWOCPU DGNQPIGF VQ VJG GCTN[ CRRNKECVKQPU QH K. lactis KP DKQVGEJPQNQI[. 9JG[ KU C NCEVQUGÿTKEJ YCUVG RTQFWEV QH FCKT[ KPFWUVT[ YJKEJ KU WUGF CU C ECTDQP UQWTEG KP K. lactisÿ OGFKCVGF DKQEQPXGTUKQP RTQEGUUGU þ)JCN[ GV CN. üùù3÷ /CWNNW GV CN. üùùùý. úCEVQUG KU VCMGP WR D[ C URGEKHKE RGTOGCUG CPF J[FTQN[UGF KPVTCEGNNWNCTN[ D[ C "ÿICNCEVQUKFCUG KPVQ INWEQUG CPF ICNCEVQUG þ4KNG[ GV CN. üùõ7 CPF TGHGTGPEGU VJGTGKPý. úCEVQUG OGVCDQNKUO KU KPFWEKDNG D[ NCEVQUG QT ICNCEVQUG KP VJG OGFKWO. +PFWEVKQP D[ NCEVQUG TGSWKTGU UWICT WRVCMG CPF J[FTQN[UKU UKPEG KPVTCEGNNWNCT ICNCEÿ VQUG UGTXGU CU VJG KPFWEKPI OQNGEWNG þUGG DGNQYý. )CNCEVQUG KU OGVCDQNKUGF D[ VJG GP\[OGU QH VJG úGNQKT RCVJYC[, GPEQFGF D[ VJG IGPGU GAL1, GAL10, CPF GAL7, YJKEJ CTG EQÿTGIWNCVGF YKVJ VJG ¾ÿICNCEVQUKFCUG CPF NCEVQUG RGTOGCUG IGPGU LAC4 CPF LAC12 IGPGU þ4KNG[ CPF &KEMUQP, üùõòý.
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üõù
#P WPWUWCNN[ NCTIG û.ø MD FKXGTIGPV EQPVTQN TGIKQP UGRCTCVGU LAC4 CPF LAC12 CPF DKPFU TGIWNCVQT[ RTQVGKPU YJKEJ EQÿQTFKPCVGN[ EQPVTQN GZRTGUUKQP QH DQVJ IGPGU D[ EQORNGZ RTQVGKPÿRTQVGKP KPVGTCEVKQPU þúGQPCTFQ GV CN. üùõ7÷ ){FGEMG GV CN. üùùü÷ -TKLIGT CPF $TGWPKI, WPRWDNKUJGF FCVCý. 6JG TGIWNCVQT TGURQPUKDNG HQT VTCPUETKRVKQP CEVKXCVKQP KP TGURQPUG VQ ICNCEVQUG YCU UJQYP VQ DG VJG JQOQNQI QH )CNò, PCOGF úCEù QT -N)CNò þ4W\\K GV CN. üùõ7÷ 5CNOGTQP CPF ìQJPUVQP, üùõø÷ 9TC[ GV CN. üùõ7÷ $TGWPKI CPF -WIGT, üùõ7ý CPF HWTVJGT TGUGCTEJ KPVQ VJG TGIWNCÿ VKQP QH VJG K. lactis LAC/GAL TGIWNQP D[ VJKU OQNGEWNG ITGCVN[ DGPGHKVGF HTQO VJG UKOKNCTKV[ VQ VJG S. cerevisiae U[UVGO þEQOR. 4GGEG CPF 2NCVV, üùù7÷ 5EJCHHTCVJ CPF $TGWPKI, ûööö HQT TGEGPV TGXKGYUý. &GURKVG VJG HCEV VJCV Klgal4 OWVCPV ECP DG EQORNGOGPVGF D[ VJG S. cerevisiae GAL4 IGPG CPF vice versa þ4KNG[ GV CN. üùõ7÷ 5CNOGTQP CPF ìQJPUVQP, üùõøý, VJGTG KU C TGOCTMCDNG FGITGG QH URGEKGU URGEKHKEKV[ KP VJG VTCPUETKRVKQPCN UYKVEJ VJCV TGIWÿ NCVGU LAC/GAL IGPG GZRTGUUKQP. #EVWCNN[, VJG NCEM QH TGFWPFCPE[ KP VJG K. lactis IGPQOG RTQXKFGF C ENWG CU VQ JQY )CNò HWPEVKQP KU CEVKXCVGF KP TGURQPUG VQ ICNCEÿ VQUG þ/G[GT GV CN. üùùü÷
üùö
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
úCEVQUG CU QRRQUGF VQ ICNCEVQUG KU CP CDWPFCPV UWICT KP VJG PCVWTCN JCDKVCV QH K. lactis, YJKEJ OC[ GZRNCKP YJ[ S. cerevisiae CPF K. lactis JCXG FKXGTIGF KP VJG TGIWNCVKQP QH VJG LAC/GAL TGIWNQP. 5KPEG KPVTCEGNNWNCT ICNCEVQUG KU VJG KPFWEGT, VJG TGIWNCVKQP QH UWICT WRVCMG RNC[U CP KORQTVCPV TQNG KP VTCPUETKRVKQPCN EQPVTQN QH LAC/GAL IGPG GZRTGUUKQP. úCEVQUG WRVCMG D[ VJG NCEVQUG RGTOGCUG QEEWTU D[ RTQÿ VQP U[ORQTV, YJKEJ CNNQYU HQT KPVTCEGNNWNCT CEEWOWNCVKQP QH VJG UWICT þ&KEMUQP CPF $CTT üùõ3ý. )CNCEVQUG ECP DG VTCPURQTVGF D[ VJG UCOG RGTOGCUG DWV, KP CFFKÿ VKQP, QVJGT VTCPURQTVGTU DGNQPIKPI VQ VJG JGZQUG VTCPURQTVGT UWDHCOKN[ QH OCLQT HCÿ EKNKVCVQTU ECP OGFKCVG ICNCEVQUG WRVCMG FQYP C EQPEGPVTCVKQP ITCFKGPV. 6JG JGZQUG VTCPURQTVGT 4CIü KU KPFWEGF D[ XCTKQWU JGZQUGU KPENWFKPI ICNCEVQUG CPF OC[ DG QPG QH VJGUG þ2TKQT GV CN. üùù3Dý. +P CFFKVKQP, CU OGPVKQPGF CDQXG, C PGY ICNCEVQUG CPF INWEQUG VTCPURQTVGT IGPG, KHT3, JCU TGEGPVN[ DGGP ENQPGF. 5KPEG VJG LAC12 IGPG KU GZRTGUUGF CV C XGT[ NQY TCVG KP PQPÿKPFWEGF EGNNU þ){FGEMG GV CN. üùùüý, VJGUG QVJGT VTCPURQTVGTU OC[ RNC[ CP KORQTVCPV TQNG KP VJG KPKVKCVKQP QH KPFWEVKQP. 6JG IGPGU LAC4 CPF LAC12 JCXG DGGP VTCPUHGTTGF VQ S. cerevisiae CPF YGTG UJQYP VQ DG UWHHKEKGPV VQ CNNQY HQT ITQYVJ QP NCEVQUG þ4WDKQÿ6GZGKTC GV CN. ûööüý. *QYGXGT, OQUV S. cerevisiae VTCPUHQTOCPVU UJQYGF UNQY ITQYVJ TCVGU CPF RQQT GZRTGUUKQP QH VJG LAC IGPGU þ5TGGMTKUJPC CPF &KEMUQP üùõôý CPF VJG HGY VTCPUÿ HQTOCPVU VJCV ITGY HCUVGT EQPVCKPGF OWNVKRNG KPVGITCVGF EQRKGU QH LAC4-LAC12 IGPGU. 5KPEG 5E)CNòR ECP CEVKXCVG VJG LAC IGPGU KP K. lactis, VJG RQQT GZRTGUUKQP QH K. lactis LAC IGPGU KP S. cerevisiae KU RTQDCDN[ FWG VQ OGEJCPKUOU VJCV TGUVTKEV CEEGUU QH )CNòR VQ VJG DKPFKPI UKVGU KP VJG LAC4-LAC12 RTQOQVGT þúCPIJCOOGT CPF $TGWPKI, WPRWDNKUJGFý. 6.3.3 ÿJG RGVKVGýPGICVKXG RJGPQV[RG /GVCDQNKUO QH K. lactis KU RTGFQOKPCPVN[ TGURKTCVQT[ CPF VJG SWGUVKQP KU YJGVJGT VJKU [GCUV ECP ITQY KP VJG EQORNGVG CDUGPEG QH QZ[IGP. 9JGTGCU, KV KU IGPGTCNN[ CEEGRVGF VJCV K. lactis FQGU PQV ITQY WPFGT UVTKEV CPCGTQDKE EQPFKVKQPU þ5WDsM GV CN. üù7ò÷ 8KUUGT GV CN. üùùö÷ 6To\oIWGV GV CN. üùùùý, QVJGTU JCXG TGRQTVGF ITQYVJ þ©PVKCP CPF $CTPGVV üùõ3ý QT GXGP CDWPFCPV ITQYVJ þ)QP\hNG\ 5KUQ GV CN. üùùøý WPFGT UWEJ EQPFKVKQPU. &GURKVG VJKU EQPVTQXGTU[, KV JCU DGGP HKTON[ GUVCDNKUJGF VJCV K. lactis KU ÉRGVKVGÿ PGICVKXGÌ. +P S. cerevisiae UOCNN EQNQPKGU, RGVKVGU, CU QRRQUGF VQ ITCPFGU, CRRGCT URQPVCPGQWUN[ FWG VQ NCTIG FGNGVKQPU KP QT GXGP EQORNGVG NQUU QH VJG OKVQEJQPFTKCN IGPQOG. %QPUGSWGPVN[, TGURKTCVKQP KU NQUV CPF RGVKVGU ECPPQV ITQY QP PQPÿ HGTOGPVCDNG ECTDQP UQWTEGU. 6JG HTGSWGPE[ QH RGVKVGU ECP DG TCKUGF UKIPKHKECPVN[ D[ GZRQUWTG QH VJG EGNNU VQ KPVGTECNCVKPI CIGPVU UWEJ CU GVJKFKWO DTQOKFG. ©XGP VJG WUG QH UWEJ CIGPVU FQGU PQV NGCF VQ VJG CRRGCTCPEG QH UVCDNG RGVKVGU KP K. lactis. 6JKU KU UQOGYJCV UWTRTKUKPI UKPEG VJKU [GCUV KU CDNG VQ HGTOGPV CPF RTQFWEG GVJCPQN CPF VJG 4CIï UVTCKPU ECP ITQY KP VJG CDUGPEG QH TGURKTCVKQP YJGP VJKU KU DNQEMGF D[ CPVKO[EKP # þ9oUQNQYUMKÿúQWXGN GV CN. üùùûCý QT DNQEMGF D[ FGNGVKQP QH VJG WPKSWG KlCYC1 IGPG þ%JGP CPF %NCTMÿ9CNMGT üùù3ý. *QYGXGT, OWVCVKQPU KP GKVJGT VJG ½, ¾, QT Ä UWDWPKVU QH VJG (ü UWDWPKV QH VJG OKÿ VQEJQPFTKCN (ö(üÿ#62 U[PVJCUG OC[ ECWUG C RGVKVGÿRQUKVKXG RJGPQV[RG. ðRQP
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üùü
VTGCVOGPV YKVJ GVJKFKWO DTQOKFG, CP[ QH VJGUG OWVCPVU TGCFKN[ HQTOU Ðö RGVKVGU. *QY VJGUG OWVCVKQPU UWRRTGUU Ðö NGVJCNKV[ KU WPENGCT. 6JG (ö UWDWPKV FQGU PQV CRÿ RGCT VQ RNC[ C TQNG CPF VJG CDGTTCPV (ü UWDWPKV ECP UVKNN DG HQTOGF KP VJG CDUGPEG QH (ö þ%JGP GV CN. üùùõ CPF TGHGTGPEGU VJGTGKPý. 6JG OWVCPVU CTG TGURKTCVQT[ EQORGÿ VGPV, DWV NQUG VJKU CDKNKV[ KP VJG RTGUGPEG QH CP QVJGTYKUG ET[RVKE OWVCVKQP þNKR$ÿüý KP VJG LIPB IGPG, GPEQFKPI C NKRQ[NÿRTQVGKP NKICUG KPXQNXGF KP VJG NKRQ[NCVKQP QH RTQVGKPU þ%JGP üùù7ý. +P S.cerevisiae, VYQ IGPGU CTG MPQYP YJKEJ CTG TGSWKTGF HQT VJG RGVKVGÿRQUKVKXG RJGPQV[RG, K.G. OP1/AAC2 QPG QH VJTGG OKVQEJQPFTKCN #62ñ#&2 VTCPUNQECVQT IGPGU CPF PEL1/PGS1 GPEQFKPI RJQURJCVKF[NIN[EGTQNRJQURJCVG U[PVJCUG KPÿ XQNXGF KP ECTFKQNKRKP DKQU[PVJGUKU. $QVJ op1/aac2 CPF pel1/pgs1 OWVCVKQPU TGPFGT S. cerevisiae RGVKVGÿPGICVKXG þ)DGNUMC GV CN. üùùùý. +P EQPVTCUV VQ S.cerevisiae, K. lactis JCU C UKPING OKVQEJQPFTKCN #62ñ#&2 VTCPUNQECVQT þ8KQNC GV CN. üùùôý CPF FKUTWRVKQP QH VJG IGPG OCMGU VJG OWVCPVU TGURKTCVQT[ FGHKEKGPV CPF ECWUGU CP KPÿ CDKNKV[ VQ ITQY QP ICNCEVQUG, OCNVQUG, QT TCHHKPQUG. 6JG OWVCPV EQWNF DG HWNN[ EQORNGOGPVGF D[ GKVJGT ScAAC1 QT ScAAC2 þ8KQNC GV CN. üùùùý. +V YQWNF DG KPVGTÿ GUVKPI VQ UGG YJGVJGT VJG EQORNGOGPVGF UVTCKPU CTG UVKNN RGVKVGÿPGICVKXG. 5KOKNCTN[, VJG KlPEL1/PGS1 IGPG JCU TGEGPVN[ DGGP KUQNCVGF, DWV HWTVJGT TGUGCTEJ CYCKVU VJG KUQNCVKQP QH C OWVCPV þ5.6[EKCMQXC GV CN. WPRWDNKUJGFý. 6.3.4 -KNNGT 5VTCKPU #U OGPVKQPGF KP UGEVKQP ø.û.û.û, OCP[ K. lactis UVTCKPU EQPVCKP NKPGCT RNCUOKFU VJCV GPEQFG C MKNNGT VQZKP. 6JGUG UVTCKPU CTG CDNG VQ UWRRTGUU VJG ITQYVJ QH EQORGVKPI [GCUVU D[ VJG CEVKXKV[ QH VJG UGETGVGF MKNNGT VQZKP QT \[OQEKP. 6JGUG UVTCKPU CTG RTQÿ VGEVGF CICKPUV VJG VQZKP D[ CP KOOWPKV[ FGVGTOKPCPV NQECVGF QP Mü CPF RQUUKDN[ UQOG HWPEVKQP QH Mû. 6JG OGEJCPKUOU QH KOOWPKV[ CESWKUKVKQP CTG EQORNGVGN[ WPÿ MPQYP. 4GUGCTEJ KP UGXGTCN NCDU JCU HQEWUUGF QP OGEJCPKUOU VJCV TGPFGT [GCUVU UGPUKVKXG VQ VJG VQZKP CPF QP VJG KPVTCEGNNWNCT VQZKP VCTIGVU. 6JG TGUWNVU UWIIGUV VJCV VJG OQFG QH CEVKQP QH K. lactis MKNNGT VQZKP TGRTGUGPVU C PQXGNV[ COQPI OKETQDKCN VQZKPU. 5GPUKVKXG [GCUV IGPGTC KPENWFG Saccharomyces, Kluyveromyces CPF Candida DWV PQV Schizosaccharomyces þ)WPIG GV CN. üùõü÷)WPIG CPF 5CMCIWEJK, üùõüý. S. cerevisiae OWVCPVU YKVJ TGFWEGF EJKVKP EQPVGPV KP VJG EGNN YCNN CTG RCTVKCNN[ QT GPÿ VKTGN[ TGUKUVCPV CICKPUV VJG VTKOGTKE GZQVQZKP KPFKECVKPI VJCV EJKVKP HWPEVKQPU CU C FQEMKPI UKVG QT TGEGRVQT þìCDNQPQYUMK GV CN. ûööüý. +PFGGF, CNN VJTGG UWDWPKVU QH VJG GZQVQZKP ECP DG DQWPF VQ CP KOOQDKNK\GF EJKVKPÿOCVTKZ. 5KPEG VJG ½ÿUWDWPKV UJQYU UGSWGPEG UKOKNCTKV[ VQ EJKVKPCUGU, VJG RQUUKDKNKV[ GZKUVU VJCV EJKVKP FGITCFCÿ VKQP KU TGSWKTGF HQT VQZKP GPVT[ KPVQ VCTIGV EGNNU. +PVTCEGNNWNCT GZRTGUUKQP QH VJG Äÿ UWDWPKV TGUWNVU KP C ITQYVJ CTTGUV þ$WVNGT GV CN. üùùüDý. 6JGTGHQTG, CV NGCUV VJG Äÿ UWDWPKV KU NKMGN[ VQ GPVGT VJG VCTIGV EGNN. S. cerevisiae GZRTGUUKPI VJKU ÄÿUWDWPKV EQPFKVKQPCNN[ WPFGT VJG EQPVTQN QH KPFWEKDNG RTQOQVGTU UGTXGF VQ KFGPVKH[ KPVTCEGNÿ NWNCT VQZKP VCTIGVU þ616Uý. 1H VJG VQZKPÿTGUKUVCPV OWVCPVU KFGPVKHKGF KP VTCPURQUQP OWVCIGPGUKU UETGGPU C OCLQTKV[ KU FGHGEVKXG KP VJG ©NQPICVQT EQORNGZ þ(GNNQYU GV CN. ûööö÷ (TQJNQHH GV CN. ûöö3ý C UKZ UWDWPKV EQORNGZ RWTKHKGF HTQO 40# RQN[ÿ
üùû
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
OGTCUG UVCDN[ CUUQEKCVGF YKVJ EJTQOCVKP þ1VGTQ GV CN. üùùù÷ 9KPMNGT GV CN. ûööüý. &GNGVKQPU QH GCEJ QH VJG UKZ UWDWPKVU EQPHGTTGF C VQZKPÿTGUKUVCPV RJGPQV[RG CPF CPÿ QVJGT VQZ4 OWVCPV, kti12, YCU UJQYP VQ FGHKPG CP CFFKVKQPCN EQORQPGPV CV NGCUV VTCPUKGPVN[ CUUQEKCVGF YKVJ VJG EQORNGZ þ(KEJVPGT GV CN. ûööûý. 1PG QH VJG UWDWPKVU GPEQFGF D[ ELB3/TOT3 JCU JKUVQPG VTCPUHGTCUG CEVKXKV[ VJCV KU GUUGPVKCN VQ EQPHGT UGPUKVKXKV[ VQ ÄÿVQZKP. 9JGVJGT QT PQV ÄÿVQZKP DKPFU FKTGEVN[ VQ ©NQPICVQT QT TGÿ SWKTGU ©NQPICVQT HWPEVKQP HQT ITQYVJ JCU PQV DGGP UQNXGF UQ HCT. ©NQPICVQT YCU UJQYP VQ DG KPXQNXGF KP VTCPUETKRVKQP GNQPICVKQP CPF &0# TGRCKT þ5XGLUVTWR ûööûý CNVJQWIJ KV FQGU PQV CRRGCT VQ DG CUUQEKCVGF YKVJ VJG OCLQTKV[ QH GNQPICVKPI RQN[ÿ OGTCUG.
6.4. +PFWUVTKCN CRRNKECVKQPU +PFWUVTKCN KPVGTGUV HQT VJG WVKNK\CVKQP QH K.lactis UVGOU HTQO þKý VJG %TCDVTGG PGICÿ VKXG EJCTCEVGT þUGG CDQXGý, þKKý VJG HCEV VJCV HGTOGPVCVKQP VGEJPQNQI[ KU GUVCDNKUJGF HQT K. lactis UKPEG GP\[OGU, RCTVKEWNCTN[ NCEVCUG þ¾ÿICNCEVQUKFCUGý YGTG RWTKHKGF HTQO VJKU QTICPKUO QP C NCTIG UECNG, þKKKý PWOGTQWU RWDNKECVKQPU CPF RCVGPVU TGRQTVÿ KPI VJCV K. lactis KU C RTQOKUKPI JQUV HQT RTQFWEVKQP QH TGEQODKPCPV RTQVGKP þ5YKPMGNU GV CN. üùù3÷ )GNNKUUGP CPF *QNNGPDGTI üùù7÷ (NGGT, üùùûý, CPF þKXý VJG HCEV VJCV K. lactis KU VJG UGEQPF [GCUV, DGUKFGU S. cerevisiae, HQT YJKEJ VJG )4#5 þgGPGTCNN[ rGICTFGF aU sCHGý UVCVWU JCU DGGP CRRTQXGF HQT HQQF CFFKVKXGU þ$QPGMCOR CPF 1UVGTQO üùùò÷ %QGPGP GV CN. ûöööý. K. lactis JCU DGGP CRRNKGF KP C PWODGT QH DKQVGEJPQNQIKECN RTQEGUUGU. -KNNGT UVTCKPU JCXG DGGP WUGF KP OQFGN UKNCIG HGTOGPVCVKQPU CU CPVKÿ[GCUV CFFKVKXGU VQ RTGXGPV URQKNCIG D[ NCEVKE CEKFÿWVKNK\KPI [GCUV EQPVCOKPCPVU þ-KVCOQVQ GV CN. üùùùý QT KP ITCRG OWUV VQYCTFU KPFKIGPQWU [GCUV URGEKGU þ;CR GV CN. ûöööý. 2TQFWEVKQP QH NCEVKE CEKF YCU CEJKGXGF D[ KPVTQFWEKPI C DQXKPG úÿNCEVCVG FGJ[FTQIGPCUG IGPG CPF GPIKPGGTKPI R[TWXCVG OGVCDQNKUO þ2QTTQ GV CN. üùùùý. 6JG RTKOCT[ KPVGTGUV KP DKQVGEJPQNQIKECN CRRNKECVKQPU KU VQ WUG K. lactis CU C JQUV HQT TGEQODKPCPV RTQVGKP RTQFWEVKQP, FGURKVG VJG HCEV VJCV KPXGUVOGPV QH VJG DKQVGEJ KPFWUVT[ KPVQ CRRNKGF K. lactis TGUGCTEJ JCU FGENKPGF KP VJG RCUV DGECWUG QH VJG RCVÿ GPV UKVWCVKQP. #P GCTN[ RCVGPV HKNGF KP üùõõ QP "Kluyveromyces CU C JQUV UVTCKP" EQXGTGF CNN TGEQODKPCPV RTQVGKPU QDVCKPGF D[ IGPGVKE GPIKPGGTKPI KP VJKU QTICPKUO þ©2 ö 3öü ø7ö $üý. 6JKU RCVGPV JCU CNNQYGF VQ FGXGNQR K. lactis KPVQ CP KPFWUVTKCN UVTCKP HQT NCTIGÿUECNG EJ[OQUKP RTQFWEVKQP D[ )KUVÿ$TQECFGUñ&5/ÿ)KUV þ8CP FGP $GTI GV CN. üùùö÷ 5YKPMGNU GV CN. üùù3ý. 6JTQWIJ ©ðÿHWPFKPI CPF PCVKQPCN ITCPVU, TGEQODKPCPV RTQVGKP RTQFWEVKQP KP K. lactis JCU EQPVKPWGF VQ TGEGKXG EQPUKFGTCDNG CVVGPVKQP KP CECFGOKEU YJGTG OQFGN RTQVGKPU þ5YGPPGP GV CN. ûööû÷ 5CNKQNC GV CN. üùùù÷ 4QEJC GV CN. üùùø÷ )QP\CNG\ GV CN. üùù7ý CPF VQQNU YGTG FGXGNQRGF VJCV EQPÿ HKTOGF VJG RQVGPVKCN QH PQPÿEQPXGPVKQPCN [GCUVU KP IGPGTCN CPF K. lactis KP RCTVKEWÿ NCT. ðRQP FKTGEV EQORCTKUQP DGVYGGP S. cerevisiae CPF K. lactis CU JQUV U[UVGOU, VJG NCVVGT RTQXGF UWRGTKQT KP C PWODGT QH ECUGU HQT TGCUQPU VJCV CTG PQV EQORNGVGN[ WPFGTUVQQF DWV RTQDCDN[ EQTTGNCVG YKVJ VJG DCNCPEG DGVYGGP QZKFCVKXG CPF HGTOGPÿ VCVKXG OGVCDQNKUO. 6JG NCTIG TGRGTVQKTG QH RQUUKDKNKVKGU VQ KPVTQFWEG CPF GZRTGUU
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üù3
JGVGTQNQIQWU IGPGU KU CNUQ KORQTVCPV. 6JGUG KPENWFG OWNVKÿEQR[, UKPING EQR[, CPF KPVGITCVKXG XGEVQTU, EQPUVKVWVKXG QT TGIWNCVGF RTQOQVGTU, CPF UGETGVKQP UKIPCNU þTGÿ XKGYGF KP 9oUQNQYUMKÿúQWXGN GV CN. üùùø÷ )GNNKUUGP CPF *QNNGPDGTI üùù7÷ 5EJCHÿ HTCVJ CPF $TGWPKI ûöööý. +P CFFKVKQP, VJG K. lactis RTQVGQN[VKE U[UVGO þ(NQTGU GV CN. üùùùý CPF GP\[OGU VJCV KORTQXG RTQVGKP HQNFKPI þ$CQ CPF (WMWJCTC ûööüý JCXG DGGP EJCTCEVGTK\GF. 4GUGCTEJ QP VJG IN[EQU[NCVKQP U[UVGO JCU TGXGCNGF HWTVJGT FKHÿ HGTGPEGU VQ S. cerevisiae þúQRG\ÿ#XCNQU GV CN. ûööü÷ ðEEGNNGVVK GV CN. ûööö÷ ðEEGNÿ NGVVK GV CN. ûööûý. 6JG Kluyveromyces lactis IGPQOG RTQLGEV RTQOKUGU VQ GPNCTIG VJKU TGRGTVQKTG CPF VJWU GPJCPEG CRRNKECVKQP RQUUKDKNKVKGU.
üGHGTGPEGU #UVTQO 5ð, -GIGN #, 5LQUVTCPF ì1, 4KPG ì þûöööý Kluyveromyces lactis 5KTûR TGIWNCVGU ECVKQP UGPUKVKXKV[ CPF OCKPVCKPU C URGEKCNK\GF EJTQOCVKP UVTWEVWTG CV VJG ET[RVKE CNRJCÿ NQEWU. )GPGVKEU:õüÿùü #UVTQO 5ð, 4KPG ì þüùùõý 6JGOG CPF XCTKCVKQP COQPI UKNGPEKPI RTQVGKPU KP Saccharomyces cerevisiae CPF Kluyveromyces lactis. )GPGVKEU üòõ:üöûüÿüöûù $CLYC 9, 6QTEJKC 6©, *QRRGT ì© þüùõõý ;GCUV 4GIWNCVQT[ )GPG GAL3:%CTDQP 4GIWNCVKQP÷ ð#5)CN ©NGOGPVU KP %QOOQP YKVJ GAL1, GAL2, GAL7, GAL10, GAL80, CPF MEL1÷ ©PEQFGF 2TQVGKP 5VTKMKPIN[ 5KOKNCT VQ ;GCUV CPF Escherichia coli )CNCEVQMKPCUGU. /QN %GNN $KQN õ:3ò3ùÿ3òò7 $CQ 9), (WMWJCTC * þûööüý 5GETGVKQP QH JWOCP RTQVGKPU HTQO [GCUV:UVKOWNCVKQP D[ FWRNKÿ ECVKQP QH RQN[WDKSWKVKP CPF RTQVGKP FKUWNHKFG KUQOGTCUG IGPGU KP Kluyveromyces lactis. )GPG û7û:üö3ÿüüö $CTKPCIC / þüùùòý ;GCUV GP\[OG HKPFU HCOG KP NKPM VQ &0# TGRNKECVKQP. 5EKGPEG ûøô:üü7ôÿ üü7ø $GVKPC 5, )QHHTKPK 2, (GTTGTQ +, 9oUQNQYUMKÿúQWXGN / þûööüý RAG4 IGPG GPEQFGU C INWÿ EQUG UGPUQT KP Kluyveromyces lactis. )GPGVKEU üôõ:ôòüÿôòõ $KCPEJK //, 5CPVCTGNNK 4, (TQPVCNK ú þüùùüý 2NCUOKF HWPEVKQPU KPXQNXGF KP VJG UVCDNG RTQRCICVKQP QH VJG R-&ü EKTEWNCT RNCUOKF KP Kluyveromyces lactis. %WTT )GPGV üù:üôôÿ üøü $KCPEJK //, 6K\\CPK ú, &GUVTWGNNG /, (TQPVCNK ú, 9oUQNQYUMKÿúQWXGN / þüùùøý 6JG 'RGÿ VKVGÿPGICVKXG' [GCUV Kluyveromyces lactis JCU C UKPING IGPG GZRTGUUKPI R[TWXCVG FGECTÿ DQZ[NCUG CEVKXKV[. /QN /KETQDKQN üù:û7ÿ3ø $KNNCTF 2, /oPCTV 5, $NCKUQPPGCW ì, $QNQVKPÿ(WMWJCTC /, (WMWJCTC *, 9oUQNQYUMKÿúQWXGN / þüùùøý )NWEQUG WRVCMG KP Kluyveromyces lactis:4QNG QH VJG HGT1 IGPG KP INWEQUG VTCPURQTV. ì $CEVGTKQN ü7õ:ôõøöÿôõøø $NCKUQPPGCW ì, (WMWJCTC *, 9oUQNQYUMKÿúQWXGN / þüùù7ý 6JG Kluyveromyces lactis GSWKXCNGPV QH ECUGKP MKPCUG + KU TGSWKTGF HQT VJG VTCPUETKRVKQP QH VJG IGPG GPEQFKPI VJG NQYÿCHHKPKV[ INWEQUG RGTOGCUG. /QN )GP )GPGV ûô3:òøùÿò77 $QNQVKPÿ(WMWJCTC /, 6QHHCPQÿ0KQEJG %, #TVKIWGPCXG (, &WEJCVGCWÿ0IW[GP ), úGOCKTG /, /CTOGKUUG 4, /QPVTQEJGT 4, 4QDGTV %, 6GTOKGT /, 9KPEMGT 2, 9GUQNQYUMKÿúQWXGN / þûöööý )GPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU:üü. Kluyveromyces lactis. (©$5 úGVV òõ7:øøÿ7ö $QPGMCOR (ì, 1UVGTQO ì þüùùòý 1P VJG UCHGV[ QH Kluyveromyces lactis:C TGXKGY. #RRN /Kÿ ETQDKQN $KQVGEJPQN òü:üÿ3
üùò
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
$QWTICTGN &, 0IW[GP %%, $QNQVKPÿ(WMWJCTC / þüùùùý *#2ò, VJG INWEQUGÿTGRTGUUGF TGIWÿ NCVGF UWDWPKV QH VJG *#2 VTCPUETKRVKQPCN EQORNGZ KPXQNXGF KP VJG HGTOGPVCVKQPÿ TGURKTCVKQP UJKHV, JCU C HWPEVKQPCN JQOQNQIWG KP VJG TGURKTCVQT[ [GCUV Kluyveromyces lactis. /QN /KETQDKQN 3ü:üûöôÿüûüô $Q\\K #, 5CNKQNC /, (CNEQPG %, $QUUC (, /CTVKPK ( þüùù7ý 5VTWEVWTCN CPF DKQEJGOKECN UVWFÿ KGU QH CNEQJQN FGJ[FTQIGPCUG KUQ\[OGU HTQO Kluyveromyces lactis. $KQEJKO $KQRJ[U #EVC ü33ù:ü33ÿüòû $TGOGT ì þüùõ3ý %CTPKVKPG:OGVCDQNKUO CPF HWPEVKQPU. 2J[UKQN 4GX ø3:üòûöÿüòõö $TGWPKI -& þüùõùý )NWEQUG TGRTGUUKQP QH LAC IGPG GZRTGUUKQP KP [GCUV KU OGFKCVGF D[ VJG VTCPUETKRVKQPCN CEVKXCVQT ú#%ù. /QN )GP )GPGV ûüø:òûûÿòû7 $TGWPKI -&, $QNQVKPÿ(WMWJCTC /, $KCPEJK //, $QWTICTGN &, (CNEQPG %, (GTTGTQ +, (TQPÿ VCNK ú, )QHHTKPK 2, -TKLIGT ìì, /C\\QPK %, /KNMQYUMK %, 5VGGPUOC *;, 9GUQNQYUMKÿ úQWXGN /,
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üùô
%QGPGP 6/, $GTVGPU #/, FG *QQI 5%, 8GTURGGMÿ4KR %/ þûöööý 5CHGV[ GXCNWCVKQP QH C NCEVCUG GP\[OG RTGRCTCVKQP FGTKXGF HTQO Kluyveromyces lactis. (QQF %JGO 6QZKEQN 3õ:ø7üÿø77 %QORCIPQ %, $QUEJK (, &CNGHHG #, 2QTTQ &, 4CP\K $/ þüùùùý +UQNCVKQP, PWENGQVKFG UGÿ SWGPEG, CPF RJ[UKQNQIKECN TGNGXCPEG QH VJG IGPG GPEQFKPI VTKQUG RJQURJCVG KUQOGTCUG HTQO Kluyveromyces lactis. #RRN ©PXKTQPO /KETQDKQN øô:òûüøÿòûüù %\[\ /, 0CIKGE //, &KEMUQP 4% þüùù3ý #WVQTGIWNCVKQP QH GAL4 VTCPUETKRVKQP KU GUUGPVKCN HQT TCRKF ITQYVJ QH Kluyveromyces lactis QP NCEVQUG CPF ICNCEVQUG. 0WENGKE #EKFU 4GU ûü:ò37õÿò3õû &GUVTWGNNG /, /GPIJKPK 4, (TQPVCNK ú, $KCPEJK // þüùùùý 4GIWNCVKQP QH VJG GZRTGUUKQP QH VJG Kluyveromyces lactis PDC1 IGPG:ECTDQP UQWTEGÿTGURQPUKXG GNGOGPVU CPF CWVQTGIWÿ NCVKQP. ;GCUV üô:3øüÿ37ö &KEMUQP 4%, $CTT - þüùõ3ý %JCTCEVGTK\CVKQP QH úCEVQUG 6TCPURQTV KP Kluyveromyces lactis. ì $CEVGTKQN üôò:üûòôÿüûôü &QPI ì, &KEMUQP 4% þüùù7ý )NWEQUG TGRTGUUGU VJG NCEVQUGÿICNCEVQUG TGIWNQP KP Kluyveromyces lactis VJTQWIJ C SNF1 CPF MIG1-FGRGPFGPV RCVJYC[ VJCV OQFWNCVGU ICNCEVQÿ MKPCUG þGAL1ý IGPG GZRTGUUKQP. 0WEN #EKFU 4GU ûô:3øô7ÿ3øøò &QTUOCP ì%, 8CP *GGUYKLM 9%, )TKXGNN ú# þüùùöý ;GCUV IGPGTCN VTCPUETKRVKQP HCEVQT )(ü:UGSWGPEG TGSWKTGOGPVU HQT DKPFKPI VQ &0# CPF GXQNWVKQPCT[ EQPUGTXCVKQP. 0WEN #EKFU 4GU üõ:û7øùÿû77ø ©PVKCP -&, $CTPGVV ì# þüùõ3ý 5QOG IGPGVKECN CPF DKQEJGOKECN CVVGORVU VQ GNWEKFCVG VJG GPGTIGVKEU QH UWICT WRVCMG CPF GZRNCKP VJG -NW[XGT GHHGEV KP VJG [GCUV Kluyveromyces lactis. %WTT )GPGV 7:3û3ÿ3ûô (GNNQYU ì, ©TFLWOGPVÿ$TQOCIG *, 6GORUV 2, 5XGLUVTWR ìí þûöööý 6JG ©NRû UWDWPKV QH GNQPICVQT CPF GNQPICVKPI 40# RQN[OGTCUG ++ JQNQGP\[OG KU C 9&òö TGRGCV RTQVGKP. ì $KQN %JGO û7ô:üûõùøÿüûõùù (GTTGTQ +, 8KQNC #/, )QHHGCW # þüùõüý +PFWEVKQP D[ INWEQUG QH CP CPVKO[EKPÿKPUGPUKVKXG, C\KFGÿUGPUKVKXG TGURKTCVKQP KP VJG [GCUV Kluyveromyces lactis. #PVQPKG XCP úGGWYGPÿ JQGM ò7:üüÿûò (KEJVPGT ú, (TQJNQHH (, $WTMPGT -, úCTUGP /, $TGWPKI -&, 5EJCHHTCVJ 4 þûööûý /QNGEWNCT CPCN[UKU QH KTI12/TOT4, C Saccharomyces cerevisiae IGPG TGSWKTGF HQT Kluyveromyces lactis \[OQEKP CEVKQP. /QN /KETQDKQN ò3:7õ3ÿ7ùü (NGGT 4 þüùùûý ©PIKPGGTKPI [GCUV HQT JKIJ NGXGN GZRTGUUKQP. %WTT 1RKP $KQVGEJPQN 3:òõøÿ òùø (NKMYGGTV /6,
üùø
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
(TGKTGÿ2KEQU /#, 4QFTKSWG\ÿ6QTTGU #/, 4COKN ©, %GTFCP /©, $TGWPKI -&, *QNNGPDGTI %2,
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üù7
)QP\hNG\ÿ&QOKPIWG\ /, (TGKTGÿ2KEQU /#, 4COKN ©, )WKCTF $, %GTFhP /© þûöööý *GOGÿ OGFKCVGF VTCPUETKRVKQPCN EQPVTQN KP Kluyveromyces lactis. %WTT )GPGV 3õ:ü7üÿü77 )QP\hNG\ÿ&QOKPIWG\ /, /oPFG\ %CTTQ %, %GTFhP /© þüùù7ý +UQNCVKQP CPF EJCTCEVGTK\Cÿ VKQP QH VJG KlHEM1 IGPG KP Kluyveromyces lactis.;GCUV ü3:ùøüÿù7ü )QP\hNG\ 5KUQ /+, (TGKTG 2KEQU /#, %GTFhP /© þüùùøý 4GQZKFCVKQP QH VJG 0#&2* RTQÿ FWEGF D[ VJG RGPVQUG RJQURJCVG RCVJYC[ KU PGEGUUCT[ HQT VJG WVKNK\CVKQP QH INWEQUG D[ Kluyveromyces lactis rag2 OWVCPVU. (©$5 úGVV 3õ7:7ÿüö )QP\hNG\ 5KUQ /+, 4COKN ©, %GTFhP /©, (TGKTGÿ2KEQU /# þüùùøý 4GURKTQHGTOGPVCVKXG OGÿ VCDQNKUO KP Kluyveromyces lactis:©VJCPQN RTQFWEVKQP CPF VJG %TCDVTGG GHHGEV. ©P\[OG /KETQDKCN 6GEJPQN üõ:ôõôÿôùü )TCJCO +4, *CY 4#, 5RKPM -), *CNFGP -#, %JCODGTU # þüùùùý +P XKXQ CPCN[UKU QH HWPEÿ VKQPCN TGIKQPU YKVJKP [GCUV 4CRüR. /QN %GNN $KQN üù:7òõüÿ7òùö )WKNNGP ©, #DGKLQP %, *KTUEJDGTI %$ þüùùùý 6JG IGPGU HQT VJG )QNIK CRRCTCVWU 0ÿ CEGV[NINWEQUCOKP[NVTCPUHGTCUG CPF VJG ð&2ÿ0ÿCEGV[NINWEQUCOKPG VTCPURQTVGT CTG EQPÿ VKIWQWU KP Kluyveromyces lactis. ì $KQN %JGO û7ò:øøòüÿøøòø )WPIG 0 þüùõøý úKPGCT &0# MKNNGT RNCUOKFU HTQO VJG [GCUV Kluyveromyces. ;GCUV û:üô3ÿ üøû )WPIG 0, 5CMCIWEJK - þüùõüý +PVGTIGPGTKE VTCPUHGT QH FGQZ[TKDQPWENGKE CEKF MKNNGT RNCUÿ OKFU, R)-Nü CPF R)-Nû, HTQO Kluyveromyces lactis KPVQ Saccharomyces cerevisiae D[ EGNN HWUKQP. ì $CEVGTKQN üò7:üôôÿüøö )WPIG 0, 6COCTW #, 1\CYC (, 5CMCIWEJK - þüùõüý +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH NKPGCT FGQZ[TKDQPWENGKE CEKF RNCUOKFU HTQO Kluyveromyces lactis CPF VJG RNCUOKFÿCUUQEKCVGF MKNNGT EJCTCEVGT. ì $CEVGTKQN üòô:3õûÿ3ùö *CY 4, ;CTTCIWFK #&, ðGOWTC * þûööüý +UQNCVKQP QH GCR1, C OCLQT VTCPUETKRVKQP HCEVQT QH IN[EQN[VKE IGPGU KP Saccharomyces cerevisiae, HTQO Kluyveromyces lactis. ;GCUV üõ:7ûùÿ73ô *GKPKUEJ ì, -KTEJTCVJ ú, úKGUGP 6, 8QIGNUCPI -, *QNNGPDGTI %2 þüùù3ý /QNGEWNCT IGPGVKEU QH RJQURJQHTWEVQMKPCUG KP VJG [GCUV Kluyveromyces lactis. /QN /KETQDKQN õ:ôôùÿô7ö *GTOCP #, 4QOCP * þüùøøý #NNGNG URGEKHKE FGVGTOKPCPVU QH JQOQVJCNNKUO KP Saccharomyces lactis. )GPGVKEU ô3:7û7ÿ7òö *GWU ìì,
üùõ
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
-KGTU ì,
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
üùù
úQFK 6, 5CNKQNC /, &QPPKPK %, )QHHTKPK 2 þûööüý 6JTGG VCTIGV IGPGU HQT VJG VTCPUETKRVKQPCN CEVKXCVQT %CVõR QH Kluyveromyces lactis:#EGV[N EQGP\[OG # U[PVJGVCUG IGPGU KLACS1 CPF KlACS2 CPF NCEVCVG RGTOGCUG IGPG KlJEN1. ì $CEVGTKQN üõ3:ôûô7ÿôûøü úQRG\ÿ#XCNQU /&, ðEEGNNGVVK &, #DGKLQP %, *KTUEJDGTI %$ þûööüý 6JG ð&2CUG CEVKXKV[ QH VJG Kluyveromyces lactis )QNIK )&2CUG JCU C TQNG KP WTKFKPG PWENGQVKFG UWICT VTCPURQTV KPVQ )QNIK XGUKENGU. )N[EQDKQNQI[ üü:òü3ÿòûû úW[VGP -, FG -QPKPI 9, 6GUUGWT +, 4WK\ /%, 4COQU ì, %QDDCGTV 2, 6JGXGNGKP ì/, *QJÿ OCPP 5 þüùù3ý &KUTWRVKQP QH VJG Kluyveromyces lactis GGS1 IGPG ECWUGU KPCDKNKV[ VQ ITQY QP INWEQUG CPF HTWEVQUG CPF KU UWRRTGUUGF D[ OWVCVKQPU VJCV TGFWEG UWICT WRVCMG. ©WT ì $KQEJGO ûü7:7öüÿ7ü3 /CWNNW %, úCORKU ), $CUKNG 6, +PIKCPPK #, 4QUUQNKPK )/, 2QORGK 4 þüùùùý 2TQFWEVKQP QH N[UQ\[OGÿGPTKEJGF DKQOCUU HTQO EJGGUG KPFWUVT[ D[ÿRTQFWEVU. ì #RRN /KETQDKQN õø:üõûÿüõø /E©CEJGTP /ì, -TCWUMQRH #, $NCEMDWTP ©* þûöööý 6GNQOGTGU CPF VJGKT EQPVTQN. #PPW 4GX )GPGV 3ò:33üÿ3ôõ /E©CEJGTP /ì, ðPFGTYQQF &*, $NCEMDWTP ©* þûööûý &[PCOKEU QH VGNQOGTKE &0# VWTPÿ QXGT KP [GCUV. )GPGVKEU üøö:ø3ÿ73 /E0GGN &), 6COCPQK ( þüùùüý 6GTOKPCN TGIKQP TGEQIPKVKQP HCEVQT ü, C &0#ÿDKPFKPI RTQÿ VGKP TGEQIPK\KPI VJG KPXGTVGF VGTOKPCN TGRGCVU QH VJG R)-N NKPGCT &0# RNCUOKFU. 2TQE 0CVN #ECF 5EK ð 5 # õõ:üü3ùõÿüüòöû /GKPJCTFV (, 5EJCHHTCVJ 4, úCTUGP / þüùù7ý /KETQDKCN NKPGCT RNCUOKFU. #RRN /KETQDKQN $KQVGEJPQN ò7:3ûùÿ33ø /GNNQT ì, ìKCPI 9, (WPM /, 4CVJLGP ì, $CTPGU %#, *KP\ 6, *GIGOCPP ì*, 2JKNKRRUGP 2 þüùùöý %2(ü, C [GCUV RTQVGKP YJKEJ HWPEVKQPU KP EGPVTQOGTGU CPF RTQOQVGTU. ©/$1 ì ù:òöü7ÿòöûø /G[GT ì, 9CNMGTÿìQPCJ #, *QNNGPDGTI %2 þüùùüý )CNCEVQMKPCUG GPEQFGF D[ GAL1 KU C DKÿ HWPEVKQPCN RTQVGKP TGSWKTGF HQT KPFWEVKQP QH VJG GAL IGPGU KP Kluyveromyces lactis CPF KU CDNG VQ UWRRTGUU VJG gal3 RJGPQV[RG KP Saccharomyces cerevisiae. /QN %GNN $KQN üü:ôòôòÿôòøü /KNMQYUMK %, -TCORG 5, 9GKTKEJ ì, *CUUG 8, $QNGU ©, $TGWPKI -& þûööüý (GGFDCEM TGIWÿ NCVKQP QH INWEQUG VTCPURQTVGT IGPG VTCPUETKRVKQP KP Kluyveromyces lactis D[ INWEQUG WRÿ VCMG. ì $CEVGTKQN üõ3:ôûû3ÿôûûù /WNFGT 9, 5EJQNVGP +*ì/, FG $QGT 49, )TKXGNN ú# þüùùòý 5GSWGPEG QH VJG *#23 VTCPÿ UETKRVKQP HCEVQT QH Kluyveromyces lactis RTGFKEVU VJG RTGUGPEG QH C PQXGN òÿE[UVGKPG \KPEÿHKPIGT OQVKH. /QN )GP )GPGV ûòô:ùøÿüöø /WNFGT 9, 5EJQNVGP +*ì/, )TKXGNN ú# þüùùôCý %CTDQP ECVCDQNKVG TGIWNCVKQP QH VTCPUETKRÿ VKQP QH PWENGCT IGPGU EQFKPI HQT OKVQEJQPFTKCN RTQVGKPU KP Kluyveromyces lactis. %WTT )GPGV ûõ:ûø7ÿû73 /WNFGT 9, 5EJQNVGP +*, )TKXGNN ú# þüùùôDý &KUVKPEV VTCPUETKRVKQPCN TGIWNCVKQP QH C IGPG EQFKPI HQT C OKVQEJQPFTKCN RTQVGKP KP VJG [GCUVU Saccharomyces cerevisiae CPF Kluyveromyces lactis FGURKVG UKOKNCT RTQOQVGT UVTWEVWTGU. /QN /KETQDKQN ü7:õü3ÿõûò /WNFGT 9, 9KPMNGT 4#, 5EJQNVGP +*ì/,
ûöö
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
1UKPIC -#, &G *CCP /, %JTKUVKCPUQP 6, 6CDCM *( þüùõûý # PQPCPWENGQVKFG UGSWGPEG KPÿ XQNXGF KP RTQOQVKQP QH TKDQUQOCN 40# U[PVJGUKU CPF 40# RTKOKPI QH &0# TGRNKECÿ VKQP KP [GCUV OKVQEJQPFTKC. 0WENGKE #EKFU 4GU üö:7ùù3ÿõööø 1VGTQ ), (GNNQYU ì, úK ;, FG $K\GOQPV 6, &KTCE #/, )WUVCHUUQP %/, ©TFLWOGPVÿ$TQOCIG *, 6GORUV 2, 5XGLUVTWR ìí þüùùùý ©NQPICVQT, C OWNVKUWDWPKV EQORQPGPV QH C PQXGN 40# RQN[OGTCUG ++ JQNQGP\[OG HQT VTCPUETKRVKQPCN GNQPICVKQP. /QN %GNN 3:üöùÿüüõ 1XGTMCOR -/, $CMMGT $/, 5VGGPUOC *;, 8CP &KLMGP ì2, 2TQPM ì6 þûööûý 6YQ OGEJCÿ PKUOU HQT QZKFCVKQP QH E[VQUQNKE 0#&2* D[ Kluyveromyces lactis OKVQEJQPFTKC. ;GCUV üù:õü3ÿõûò 1\KGTÿ-CNQIGTQRQWNQU 1, /CNRGTVW[ #, $Q[GT ì, 6GMCKC (, &WLQP $ þüùùõý 4CPFQO GZRNQÿ TCVKQP QH VJG Kluyveromyces lactis IGPQOG CPF EQORCTKUQP YKVJ VJCV QH Saccharomyces cerevisiae. 0WENGKE #EKFU 4GU ûø:ôôüüÿôôûò 2GPI ), *QRRGT ì© þûöööý ©XKFGPEG HQT )CN3R'U E[VQRNCUOKE NQECVKQP CPF )CNõöR'U FWCN E[ÿ VQRNCUOKEÿ PWENGCT NQECVKQP KORNKECVGU PGY OGEJCPKUOU HQT EQPVTQNNKPI )CNòR CEVKXKV[ KP Saccharomyces cerevisiae. /QN %GNN $KQN ûö:ôüòöÿôüòõ 2KUMWT ì þûööüý 1TKIKP QH VJG FWRNKECVGF TGIKQPU KP VJG [GCUV IGPQOGU. 6TGPFU )GPGV ü7:3öûÿ3ö3 2NCVV #, 4QUU *%, *CPMKP 5, 4GGEG 4ì þûöööý 6JG KPUGTVKQP QH VYQ COKPQ CEKFU KPVQ C VTCPÿ UETKRVKQPCN KPFWEGT EQPXGTVU KV KPVQ C ICNCEVQMKPCUG. 2TQE 0CVN #ECF 5EK ð 5 # ûööö /CT ûõ÷ ù7þ7ý:3üôòÿù ù7:3üôòÿ3üôù 2QTTQ &, $KCPEJK //, $TCODKNNC ú, /GPIJKPK 4, $QN\CPK &, %CTTGTC 8, úKGXGPUG ì, úKW %ú, 4CP\K $/, (TQPVCNK ú, #NDGTIJKPC ú þüùùùý 4GRNCEGOGPV QH C OGVCDQNKE RCVJYC[ HQT NCTIGÿUECNG RTQFWEVKQP QH NCEVKE CEKF HTQO GPIKPGGTGF [GCUVU. #RRN ©PXKTQP /KETQÿ DKQN øô:òûüüÿòûüô 2TCUCF 4, /KTCNNGU 8ì, $GPKVQ 4/, 5GTTCPQ 4 þüùùùý # RTQRQUCN HQT PQOGPENCVWTG QH CNFGÿ J[FG FGJ[FTQIGPCUGU KP Saccharomyces cerevisiae CPF EJCTCEVGTK\CVKQP QH VJG UVTGUUÿ KPFWEKDNG ALD2 CPF ALD3 IGPGU. ;GCUV üô:õûùÿõòû 2TKQT %, (WMWJCTC *, $NCKUQPPGCW ì, 9oUQNQYUMKÿúQWXGN / þüùù3Cý úQYÿCHHKPKV[ INWEQUG ECTTKGT IGPG LGT1 QH Saccharomyces cerevisiae, C JQOQNQIWG QH VJG Kluyveromyces lactis RAG1 IGPG. ;GCUV ù:ü373ÿü377 2TKQT %, /COGUUKGT 2, (WMWJCTC *, %JGP îì, 9oUQNQYUMKÿúQWXGN / þüùù3Dý 6JG JGZQMKPCUG IGPG KU TGSWKTGF HQT VTCPUETKRVKQPCN TGIWNCVKQP QH VJG INWEQUG VTCPURQTVGT IGPG RAG1 KP Kluyveromyces lactis. /QN %GNN $KQN ü3:3õõûÿ3õõù 2TKQT %, 6K\\CPK ú, (WMWJCTC *, 9oUQNQYUMKÿúQWXGN / þüùùøý RAG3 IGPG CPF VTCPUETKRÿ VKQPCN TGIWNCVKQP QH VJG R[TWXCVG FGECTDQZ[NCUG IGPG KP Kluyveromyces lactis. /QN /Kÿ ETQDKQN ûö:7øôÿ77û 4CIPKPK #, (WMWJCTC * þüùõõý /KVQEJQPFTKCN &0# QH VJG [GCUV Kluyveromyces:IWCPKPGÿ E[VQUKPG TKEJ UGSWGPEG ENWUVGTU. 0WENGKE #EKFU 4GU üø:õò33ÿõòòû 4COKN ©, (TGKTGÿ2KEQU /#, %GTFhP /© þüùùõý %JCTCEVGTK\CVKQP QH RTQOQVGT TGIKQPU KPÿ XQNXGF KP JKIJ GZRTGUUKQP QH KlCYC1. ©WT ì $KQEJGO ûôø:ø7ÿ7ò 4GGEG 4ì, 2NCVV # þüùù7ý 5KIPCNNKPI CEVKXCVKQP CPF TGRTGUUKQP QH 40# RQN[OGTCUG ++ VTCPÿ UETKRVKQP KP [GCUV. $KQ©UUC[U üù:üööüÿüöüö 4GNKGPG 4, 5CUPCWMCU - þüùù7ý *GVGTQNQIQWU IGPG GZRTGUUKQP QP VJG NKPGCT RNCUOKFU QH Kluyveromyces lactis. ;GCUV ü3:5ûû7 4KNG[ /+, &KEMUQP 4% þüùõòý )GPGVKE CPF DKQEJGOKECN EJCTCEVGTK\CVKQP QH VJG ICNCEVQUG IGPG ENWUVGT KP Kluyveromyces lactis. ì $CEVGTKQN üôõ:7öôÿ7üû
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
ûöü
4KNG[ /+, 5TGGMTKUJPC 5, $JCKTK 5, &KEMUQP 4% þüùõ7ý +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH OWÿ VCPVU KP Kluyveromces lactis FGHGEVKXG KP NCEVQUG VTCPURQTV. /QN )GP )GPGV ûöõ:üòôÿ üôü 4QEJC 6ú, 2CVGTUQP ), %TKOOKPU -, $Q[F #, 5CY[GT ú, (QVJGTIKNNÿ)KNOQTG ú# þüùùøý ©ZRTGUUKQP CPF UGETGVKQP QH TGEQODKPCPV QXKPG DGVCÿNCEVQINQDWNKP KP Saccharomyces cerevisiae CPF Kluyveromyces lactis. $KQEJGO ì 3ü3 þ 2V 3ý:ùû7ÿù3û 4QOCPQU /#, $Q[F # þüùõõý # VTCPUETKRVKQPCN DCTTKGT VQ GZRTGUUKQP QH ENQPGF VQZKP IGPGU QH VJG NKPGCT RNCUOKF Mü QH Kluyveromyces lactis:GXKFGPEG VJCV PCVKXG Mü JCU PQXGN RTQOQVGTU. 0WENGKE #EKFU 4GU üø:7333ÿ73ôö 4WDKQÿ6GZGKTC /, #TGXCNQÿ4QFTKIWG\ /, úGSWGTKEC ìú, 2QNCKPC ì þûööüý úCEVQUG WVKNK\CVKQP D[ Saccharomyces cerevisiae UVTCKPU GZRTGUUKPI Kluyveromyces lactis LAC IGPGU. ì $KQVGEJPQN õò:ù7ÿüöø 4W\\K /, $TGWPKI -&, (KEEC #), *QNNGPDGTI %2 þüùõ7ý 2QUKVKXG TGIWNCVKQP QH VJG ¾ÿ ICNCEVQUKFCUG IGPG HTQO Kluyveromyces lactis KU OGFKCVGF D[ CP WRUVTGCO CEVKXCVKQP UKVG VJCV UJQYU JQOQNQI[ VQ VJG GAL WRUVTGCO CEVKXCVKQP UKVG QH Saccharomyces cerevisiae. /QN %GNN $KQN 7:ùùûÿùù7 5CFQYUMK +, %QUCV %, &JCPCYCPUC 4 þüùùøý 2JQURJQT[NCVKQP QH )CNòR CV C UKPING %ÿ VGTOKPCN TGUKFWG KU PGEGUUCT[ HQT ICNCEVQUGÿKPFWEKDNG VTCPUETKRVKQP. /QN %GNN $KQN üø:òõ7ùÿòõõ7 5CNKQNC /, /C\\QPK %, 5QNKOCPFQ 0, %TKUC #, (CNEQPG %, ìWPI ), (NGGT 4 þüùùùý ðUG QH VJG KlADH4 RTQOQVGT HQT GVJCPQNÿFGRGPFGPV RTQFWEVKQP QH TGEQODKPCPV JWOCP UGTWO CNDWOKP KP Kluyveromyces lactis. #RRN ©PXKTQPO /KETQDKQN øô:ô3ÿøö 5CNOGTQP ì/, ìQJPUVQP 5# þüùõøý #PCN[UKU QH VJG Kluyveromyces lactis RQUKVKXG TGIWNCVQT[ IGPG LAC9 TGXGCNU HWPEVKQPCN JQOQNQI[ VQ, DWV UGSWGPEG FKXGTIGPEG HTQO, VJG Saccharomyces cerevisiae GAL4 IGPG. 0WENGKE #EKFU 4GU üò:77ø7ÿ77õü 5CVQ 6, úQRG\ /%, 5WIKQMC 5, ìKICOK ;, $CMGT *8, ðGOWTC * þüùùùý 6JG ©ÿDQZ &0# DKPFKPI RTQVGKP 5IEüR UWRRTGUUGU VJG gcr2 OWVCVKQP, YJKEJ KU KPXQNXGF KP VTCPUETKRÿ VKQPCN CEVKXCVKQP QH IN[EQN[VKE IGPGU KP Saccharomyces cerevisiae. (©$5 úGVV òø3:3ö7ÿ 3üü 5EJCHHTCVJ 4, $TGWPKI -& þûöööý )GPGVKEU CPF OQNGEWNCT RJ[UKQNQI[ QH VJG [GCUV Kluyveromyces lactis. (WPICN )GPGV $KQN 3ö:ü73ÿüùö 5EJCHHTCVJ 4, /GCEQEM 2# þûööüý #P 55$ GPEQFGF D[ CPF QRGTCVKPI QP NKPGCT MKNNGT RNCUÿ OKFU HTQO Kluyveromyces lactis. ;GCUV üõ:üû3ùÿüûò7 5EJCHHTCVJ 4, /GKPJCTFV (, /GCEQEM 2# þüùù7ý 14(7 QH [GCUV RNCUOKF R)-úû:CPCN[UKU QH IGPG GZRTGUUKQP KP XKXQ. %WTT )GPGV 3ü:üùöÿüùû 5EJCHHTCVJ 4, /GKPJCTFV (, /GCEQEM 2# þüùùùý )GPGVKE OCPKRWNCVKQP QH Kluyveromyces lactis NKPGCT &0# RNCUOKFU:IGPG VCTIGVKPI CPF RNCUOKF UJWHHNGU. (©/5 /KETQDKQN úGVV ü7õ:ûöüÿûüö 5EJCHHTCVJ 4, 5QQPF 5/, /GCEQEM 2# þüùùôý 6JG &0# CPF 40# RQN[OGTCUG IGPGU QH [GCUV RNCUOKF R)-úû CTG GUUGPVKCN NQEK HQT RNCUOKF KPVGITKV[ CPF OCKPVGPCPEG. /KETQÿ DKQNQI[ üòü þ 2V üöý:ûôùüÿûôùù 5EJKEMGN ì, *GNOKI %, /GKPJCTFV ( þüùùøý Kluyveromyces lactis MKNNGT U[UVGO:CPCN[UKU QH E[VQRNCUOKE RTQOQVGTU QH VJG NKPGCT RNCUOKFU. 0WENGKE #EKFU 4GU ûò:üõ7ùÿüõõø 5GQKIJG %, 9QNHG -* þüùùùý ;GCUV IGPQOG GXQNWVKQP KP VJG RQUVÿIGPQOG GTC. %WTT 1RKP /KETQDKQN û:ôòõÿôôò 5QWEKGV ì, #KING /, #TVKIWGPCXG (, $NCPFKP ), $QNQVKPÿ(WMWJCTC /, $QP ©, $TQVVKGT 2, %CUCTGIQNC 5, FG /QPVKIP[ ì, &WLQP $, &WTTGPU 2, )CKNNCTFKP %, úGRKPING #, úNQTGPVG $, /CNRGTVW[ #, 0GWXGINKUG %, 1\KGTÿ-CNQIGTQRQWNQU 1, 2QVKGT 5, 5CWTKP 9, 6GMCKC (,
ûöû
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
6QHHCPQÿ0KQEJG %, 9GUQNQYUMKÿúQWXGN /, 9KPEMGT 2, 9GKUUGPDCEJ ì þûöööý )Gÿ PQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU:ü. # UGV QH [GCUV URGEKGU HQT OQÿ NGEWNCT GXQNWVKQP UVWFKGU. (©$5 úGVV òõ7:3ÿüû 5TGGMTKUJPC -, &KEMUQP 4% þüùõôý %QPUVTWEVKQP QH UVTCKPU QH Saccharomyces cerevisiae VJCV ITQY QP NCEVQUG. 2TQE 0CVN #ECF 5EK ð5# õû:7ùöùÿ7ùü3 5VCTM /ì, $Q[F # þüùõøý 6JG MKNNGT VQZKP QH Kluyveromyces lactis:%JCTCEVGTK\CVKQP QH VJG VQZKP UWDWPKVU CPF KFGPVKHKECVKQP QH VJG IGPGU YJKEJ GPEQFG VJGO. ©/$1 ì ô:üùùôÿ ûööû 5VCTM /ì, $Q[F #, /KNGJCO #ì, 4QOCPQU /# þüùùöý 6JG RNCUOKFÿGPEQFGF MKNNGT U[UVGO QH Kluyveromyces lactis:C TGXKGY. ;GCUV ø:üÿûù 5VGGPUOC *;, FG ìQPIJ (%/, úKPPGMCOR / þüùõõý 6JG WUG QH GNGEVTQRJQTGVKE MCT[QV[RGU KP VJG ENCUUKHKECVKQP QH [GCUVU:Kluyveromyces marxianus CPF K.lactis. %WTT )GPGV üò:3üüÿ3ü7 5VGGPUOC *;, 6GT úKPFG ìì/ þûööüý 2NCUOKFU YKVJ VJG %TGÿTGEQODKPCUG CPF VJG nat OCTMGT, UWKVCDNG HQT WUG KP RTQVQVTQRJKE UVTCKPU QH Saccharomyces cerevisiae CPF Kluyveromyces lactis. ;GCUV üõ:òøùÿò7û 5WDsM ì, -QNCTQX ì, -QXhE ú þüù7òý #PCGTQDKE ITQYVJ HQTOCVKQP QH TGURKTCVKQPÿFGHKEKGPV OWVCPVU QH XCTKQWU URGEKGU QH [GCUV. (©$5 úGVV òô:ûø3ÿûøø 5XGLUVTWR ìí þûööûý %JTQOCVKP GNQPICVKQP HCEVQTU. %WTT 1RKP )GPGV &GX üû:üôøÿüøü 5YGPPGP &, 2CWN /(, 8GTPKU ú, $GEMGTKEJ ì/, (QWTPKGT #, )CKNNCTFKP % þûööûý 5GETGVKQP QH CEVKXG CPVKÿ4CU UKPINGÿEJCKP (X CPVKDQF[ D[ VJG [GCUVU Yarrowia lipolytica CPF Kluyveromyces lactis. /KETQDKQNQI[ üòõ:òüÿôö 5YKPMGNU $9, XCP 1Q[GP #ì, $QPGMCOR (ì þüùù3ý 6JG [GCUV Kluyveromyces lactis CU CP GHHKEKGPV JQUV HQT JGVGTQNQIQWU IGPG GZRTGUUKQP. #PVQPKG 8CP úGGWYGPJQGM øò:üõ7ÿ ûöü 6CMGFC /, *KTCKUJK *, 6CMGUCMQ 6, 6CPCUG 5, )WPIG 0 þüùùøý 6JG VGTOKPCN RTQVGKP QH VJG NKPGCT &0# RNCUOKF R)-úû UJCTGU CP 0ÿ VGTOKPCN FQOCKP QH VJG RNCUOKFÿGPEQFGF &0# RQN[OGTCUG. ;GCUV üû:ûòüÿûòø 6GT úKPFG ìì/, úKCPI *, &CXKU 49, 5VGGPUOC *;, 8CP &KLMGP ì2, 2TQPM ì6 þüùùùý )Gÿ PQOGÿYKFG VTCPUETKRVKQPCN CPCN[UKU QH CGTQDKE CPF CPCGTQDKE EJGOQUVCV EWNVWTGU QH Saccharomyces cerevisiae. ì $CEVGTKQN üõü:7òöùÿ7òü3 6GT úKPFG ìì/, 5VGGPUOC *; þûöö3ý 6TCPUETKRVKQPCN TGIWNCVKQP QH YML083c WPFGT CGTQDKE CPF CPCGTQDKE EQPFKVKQPU. ;GCUV ûö:ò3ùÿòôò 6K\\CPK ú, /GCEQEM 2, (TQPVCNK ú, 9oUQNQYUMKÿúQWXGN / þüùùõý 6JG RAG3 IGPG QH Kluyveromyces lactis KU KPXQNXGF KP VJG VTCPUETKRVKQPCN TGIWNCVKQP QH IGPGU EQFKPI HQT GPÿ \[OGU KORNKECVGF KP R[TWXCVG WVKNK\CVKQP CPF IGPGU QH VJG DKQU[PVJGUKU QH VJKCOKPG R[ÿ TQRJQURJCVG. (©/5 /KETQDKQN úGVV üøõ:ûôÿ3ö 6QMWPCIC /, -CYCOWTC #, *KUJKPWOC ( þüùõùý ©ZRTGUUKQP QH R)-ú MKNNGT ûõ- UWDWPKV KP Saccharomyces cerevisiae:KFGPVKHKECVKQP QH ûõ- UWDWPKV CU C MKNNGT RTQVGKP. 0WENGKE #Eÿ KFU 4GU ü7:3ò3ôÿ3òòø 6QMWPCIC /, 9CFC 0, *KUJKPWOC ( þüùõ7ý ©ZRTGUUKQP CPF KFGPVKHKECVKQP QH KOOWPKV[ FGÿ VGTOKPCPVU QP NKPGCT &0# MKNNGT RNCUOKFU R)-úü CPF R)-úû KP Kluyveromyces lactis. 0WENGKE #EKFU 4GU üô:üö3üÿüöòø 6QOOCUKPQ / þüùùüý -KNNGT U[UVGO QH Kluyveromyces lactis:6JG QRGP TGCFKPI HTCOG üö QH VJG R)-üû RNCUOKF GPEQFGU C RWVCVKXG &0# DKPFKPI RTQVGKP. ;GCUV 7:ûòôÿûôû 6QOOCUKPQ /, 4KEEK 5, )CNGQVVK %ú þüùõõý )GPQOG QTICPK\CVKQP QH VJG MKNNGT RNCUOKF R)-üû HTQO Kluyveromyces lactis. 0WENGKE #EKFU 4GU üø:ôõø3ÿôõ7õ
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
ûö3
6To\oIWGV 8,
ûöò
-CTKP &. $TGWPKI CPF *. ;FG 5VGGPUOC
9oUQNQYUMKÿúQWXGN /, 2TKQT %, $QTPGESWG &, (WMWJCTC * þüùùûCý 4CIÿOWVCVKQPU KPÿ XQNXGF KP INWEQUG OGVCDQNKUO KP [GCUV:KUQNCVKQP CPF IGPGVKE EJCTCEVGTK\CVKQP. ;GCUV õ:7üüÿ7üù 9KNUQP &9, /GCEQEM 2# þüùõõý ©ZVTCPWENGCT IGPG GZRTGUUKQP KP [GCUV:GXKFGPEG HQT C RNCUOKFÿGPEQFGF 40# RQN[OGTCUG QH WPKSWG UVTWEVWTG. 0WENGKE #EKFU 4GU üø:õöù7ÿ õüüû 9KPMNGT ##, )QGFGIGDWTG 4*,
ø Kluyveromyces lactis: IGPGVKEU, RJ[UKQNQI[, CPF CRRNKECVKQP
ûöô
7 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris: HCXQWTCDNG EGNN HCEVQTKGU KP XCTKQWU CRRNKECVKQPU /GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
#DUVTCEV 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris CTG GZVGPÿ UKXGN[ WUGF CU OQFGN QTICPKUOU KP UVWFKGU QP RGTQZKUQOG DKQIGPGUKU CPF FGITCFCÿ VKQP. +P CFFKVKQP, VJGUG [GCUVU JCXG ICKPGF KPETGCUKPI KPVGTGUV CU EGNN HCEVQTKGU KP XCTKQWU RTQFWEVKQP RTQEGUUGU. +P VJKU EQPVTKDWVKQP, CP QXGTXKGY KU RTGUGPVGF QP UGXGTCN CURGEVU QH VJG CRRNKECVKQP QH DQVJ EGNN HCEVQTKGU. $QVJ [GCUV URGEKGU JCXG DGGP UWEEGUUHWNN[ CRRNKGF HQT VJG RTQFWEVKQP QH JGVGTQNQIQWU RTQFWEVU. 6JG OQÿ NGEWNCT VQQNU CTG JKIJN[ FGXGNQRGF, G.I. VJG CXCKNCDKNKV[ QH C NCTIG XCTKGV[ QH GZRTGUÿ UKQP XGEVQTU KP YJKEJ XCTKQWU EQPUVKVWVKXG CPF KPFWEKDNG RTQOQVGTU CTG CXCKNCDNG VQ EQPVTQN RTQVGKP RTQFWEVKQP. 6JG WUG QH DQVJ OGVJ[NQVTQRJU QHHGTU C URGEKHKE CFXCPÿ VCIG VQ CEEWOWNCVG HQTGKIP RTQVGKPU KPUKFG RGTQZKUQOGU. 6JKU KU KP RCTVKEWNCT CFÿ XCPVCIGQWU YJGP VJG RTQFWEGF RTQVGKP KU VQZKE HQT VJG [GCUV EGNN QT YJGP WPFGUKTGF OQFKHKECVKQPU (G.I. RTQVGQN[UKU QT IN[EQU[NCVKQP) JCXG VQ DG RTGXGPVGF. þWTVJGTÿ OQTG, H. polymorpha OKIJV DG VJG QTICPKUO QH EJQKEG HQT VJG RTQFWEVKQP QH IGNÿ HQTOKPI RTQVGKPU CU YGNN CU JGVGTQNQIQWU (KPENWFKPI JWOCP) OGODTCPG RTQVGKPU
7.1 +PVTQFWEVKQP /KETQQTICPKUOU JCXG CNTGCF[ DGGP GZRNQKVGF HQT SWKVG UQOG VKOG HQT HQQF RTQFWEÿ VKQP, G.I. QH YKPG, DGGT CPF DTGCF. 6JGUG RTQEGUUGU CTG IGPGTCNN[ TGICTFGF CU VTCFKÿ VKQPCN DKQVGEJPQNQI[. /QFGTP DKQVGEJPQNQI[ DGECOG HGCUKDNG YKVJ VJG CXCKNCDKNKV[ QH VJG CFXCPEGF TGEQODKPCPV &0# VGEJPQNQI[. 7UKPI VJGUG VGEJPKSWGU, OKETQQTÿ ICPKUOU PQYCFC[U CTG WUGF HQT VJG RTQFWEVKQP QH HQTGKIP, DKQNQIKECNN[ CEVKXG, RTQÿ VGKPU HQT OGFKECN, UEKGPVKHKE, CPF EQOOGTEKCN RWTRQUGU. 6Q FCVG, C XCTKGV[ QH JQUV QTICPKUOU (EGNN HCEVQTKGU) JCXG DGGP GZRNQTGF VQ RTQFWEG JGVGTQNQIQWU RTQVGKPU, TCPIKPI HTQO DCEVGTKC VQ OCOOCNKCP EGNNU. /COOCNKCP EGNNU, Escherichia coli CPF [GCUV CTG WUGF G.I. HQT VJG RTQFWEVKQP QH VJGTCRGWVKE CIGPVU YJKEJ CTG WUGF HQT VTGCVOGPV QH JGOQUVCVKE CPF VJTQODQVKE FKUQTFGTU. +P JWOCPU, FGHKEKGPE[ QH EQCIWÿ NCVKQP HCEVQT 8+++ ECWUGU VJG DNGGFKPI FKUQTFGT JGOQRJKNKC #. 6JG EWTTGPV VTGCVÿ OGPV QH VJKU FKUGCUG KU TGRNCEGOGPV VJGTCR[, WUKPI GZVGTPCN TGEQODKPCPV HCEVQT 8+++ (Tþ8+++). %WTTGPVN[, HQWT FKHHGTGPV Tþ8+++ CTG CXCKNCDNG, CNN QH YJKEJ CTG RTQFWEGF D[ TGEQODKPCPV OCOOCNKCP EGNN NKPGU ($QGFGMGT 2ýý2). 1PG QH VJG OQUV YKFGN[ Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
2ýü
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
WUGF VJTQODQN[VKE CIGPVU KU VJG VKUUWGÿV[RG RNCUOKPQIGP CEVKXCVQT (Vÿû#). # TGÿ EQODKPCPV HQTO ECNNGF TGVGRNCUG (Tû#), YJKEJ UJQYGF CP GZVGPFGF JCNHÿNKHG KP EKTÿ EWNCVKQP, KU UWEEGUUHWNN[ RTQFWEGF D[ E. coli (/CVVGU 2ýýú). 6Q FCVG, [GCUVU CTG WUGF HQT VJG JKIJÿNGXGN RTQFWEVKQP QH VJG CPVKEQCIWNCPV JKÿ TWFKP (#XIGTKPQU GV CN. 2ýýú; -KO GV CN. 2ýýú; 5QJP GV CN. 2ýýú). ùGCUVU CTG QHVGP RTGHGTTGF HQT VJG U[PVJGUKU QH TGEQODKPCPV RTQVGKPU HTQO JKIJGT GWMCT[QVGU. LKMG DCEVGTKC, [GCUV EWNVKXCVKQP KU HCUV CV NQY EQUV CPF C XCTKGV[ QH VGEJPKSWGU GZKUV HQT VJG RTQFWEVKQP CPF OCPKRWNCVKQP QH VJG RTQVGKPU RTQFWEGF. ùGCUVU, JQYGXGT, RQUUGUU URGEKHKE CFXCPVCIGU QXGT DCEVGTKCN RTQFWEVKQP U[UVGOU, CU VJG[ RTQXKFG CP GPXKTQPOGPV HQT RQUVÿVTCPUNCVKQPCN OQFKHKECVKQP CPF UGETGVKQP (G.I. CE[NCVKQP, RJQURJQT[NCVKQP, IN[EQU[NCVKQP, HQTOCVKQP QH FKUWNHKFG DQPFU). 6JGUG OQFKHKECVKQPU CTG QHVGP GUUGPVKCN HQT VJG HWPEVKQP CPFøQT UVCDKNKV[ QH VJG RTQÿ FWEGF RTQVGKP RTQFWEVU. þQT OCKPN[ JKUVQTKECN TGCUQPU, Saccharomyces cerevisiae YCU VJG HKTUV VQ DG GZRNQKVGF HQT VJG RTQFWEVKQP QH TGEQODKPCPV RTQVGKPU. 0QVYKVJÿ UVCPFKPI XCTKQWU UWEEGUUHWN CRRNKECVKQPU, URGEKHKE FKUCFXCPVCIGU QH VJG WUG QH VJKU QTICPKUO YGTG CNUQ GPEQWPVGTGF; VJGUG KPENWFG HQT KPUVCPEG NQY [KGNFU, WPFGUKTGF J[RGTIN[EQU[NCVKQP, CPF KPUVCDKNKV[ QH VJG GPIKPGGTGF RTQFWEVKQP UVTCKP. /GVJ[NQVTQRJKE [GCUVU (G.I. Hansenula polymorpha, Pichia pastoris) JCXG ICKPGF KPETGCUKPI KPVGTGUV HQT HWPFCOGPVCN TGUGCTEJ CPF CU CVVTCEVKXG JQUVU HQT VJG RTQFWEVKQP QH DKQNQIKECNN[ CEVKXG RTQVGKPU (÷GNNKUUGP GV CN. ú992; *QNNGPDGTI CPF ÷GNNKUUGP ú99ö). 6JG WUG QH VJGUG QTICPKUOU KP HWPFCOGPVCN TGUGCTEJ KU OCKPN[ TGÿ NCVGF VQ UVWFKGU QH RGTQZKUQOG JQOGQUVCUKU CPF PKVTCVG CUUKOKNCVKQP (ûGTG\ GV CN. ú99ö; 5CMCK CPF 5WDTCOCPK 2ýýý; XCP FGT -NGK CPF 8GGPJWKU 2ýý2; 8GGPJWKU GV CN. 2ýýý). # EJCTCEVGTKUVKE HGCVWTG QH RGTQZKUQOGU KP VJGUG [GCUVU KU VJCV VJGUG QTICPGNNGU CTG OCUUKXGN[ KPFWEGF FWTKPI ITQYVJ QH EGNNU QP OGVJCPQN. 7PFGT VJGUG EQPFKVKQPU, VJG QTICPGNNGU EQPVCKP VJG MG[ GP\[OGU KPXQNXGF KP OGVJCPQN OGVCDQÿ NKUO PCOGN[, CNEQJQN QZKFCUG (#1), FKJ[FTQZ[CEGVQPG U[PVCUG (&*#5), CPF ECVCÿ NCUG (%#6). +P DQVJ URGEKGU, RGTQZKUQOGU ECP CNUQ JCTDQWT GP\[OGU KPXQNXGF KP VJG OGVCDQNKUO QH GVJCPQN, RTKOCT[ COKPGU, &ÿCOKPQ CEKFU, Lÿ½ J[FTQZ[ CEKFU, CPF RWTKPGU (8GGPJWKU CPF *CTFGT ú9üü). *KIJGUV KPFWEVKQP NGXGNU QH RGTQZKUQOCN GP\[OGU CTG QDUGTXGF CV OGVJ[NQVTQÿ RJKE ITQYVJ EQPFKVKQPU. 7R VQ üýõ QH VJG E[VQRNCUOKE XQNWOG OC[ DG QEEWRKGF D[ RGTQZKUQOGU YJGP H. polymorpha KU ITQYP KP C OGVJCPQNÿNKOKVGF EJGOQUVCV (8GGPJWKU GV CN. ú9öü). +P VJGUG EGNNU, VJG VYQ RGTQZKUQOCN RTQVGKPU #1 CPF &*#5 OC[ EQPUVKVWVG QXGT ôýõ QH VQVCN EGNNWNCT RTQVGKP. 6JKU KNNWUVTCVGU VJCV VJG IGPGU GPEQFKPI VJGUG RTQVGKPU CTG EQPVTQNNGF D[ XGT[ UVTQPI RTQOQVGTU. 6JKU HGCVWTG EQPÿ VTKDWVGF VQ TGEQIPK\KPI OGVJ[NQVTQRJKE [GCUV CU CVVTCEVKXG EGNN HCEVQTKGU.
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
2ý9
7.2 6QQNU HQT VJG RTQFWEVKQP QH JGVGTQNQIQWU RTQVGKPU KP Hansenula polymorpha CPF Pichia pastoris 7.2.1 6TCPUHQTOCVKQP CPF XGEVQTU ©HHKEKGPV CPF TGNKCDNG VTCPUHQTOCVKQP RTQEGFWTGU HQT H. polymorpha CPF P. pastoris JCXG DGGP FGXGNQRGF. ùGCUV EGNNU ECP DG VTCPUHQTOGF WUKPI C URJGTQRNCUV IGPGTCVKQP OGVJQF (%GTGIJKPQ CPF %TGII 2ýýý) QT YJQNGÿEGNN OGVJQFU CEEQTFKPI VQ VJG NKVJKWO CEGVCVGÿFKOGVJ[NUWNHQZKFG OGVJQF FGUETKDGF D[ *KNN (*KNN GV CN. ú99ú), D[ CFFKPI û©÷ (&QJOGP GV CN. ú99ú) QT D[ GNGEVTQRQTCVKQP (þCDGT GV CN. ú994), QH YJKEJ VJG NCVVGT KU VJG EWTTGPV OGVJQF QH EJQKEG. #P GUUGPVKCN VQQN HQT VJG EQPUVTWEVKQP QH TGEQODKPCPV H. polymorpha CPF P. pastoris UVTCKPU CTG Eschericia coli [GCUV UJWVVNG XGEVQTU. +ORQTVCPV HGCVWTGU QH VJGUG XGEVQTU CTG VJG UGNGEVCDNG OCTMGT IGPGU, YJKEJ ECP HWPEVKQPCNN[ EQORNGOGPV XCTKÿ QWU CWZQVTQRJKE [GCUV UVTCKPU. %QOOQPN[ WUGF OCTMGT IGPGU HQT DCEVGTKCN c H. polymorpha GZRTGUUKQP XGEVQTU CTG H. polymorpha LEU2, URA3, TRP3 CPF ADE11, VJG S. cerevisiae IGPGU LEU2, URA3 CU YGNN CU Candida albicans LEU2 (#ICRJQPQX GV CN. ú994; $QIFCPQXC GV CN. ú99ó; /GTEMGNDCEJ GV CN. ú993; òQIÿ IGPMCOR GV CN. ú9üô). 6Q FCVG, CNUQ IGPGU VJCV EQPHGT TGUKUVCPEG VQYCTFU ÷4úü, RJNGQO[EKP QT \GQEKP JCXG DGGP UWEEGUUHWNN[ CRRNKGF CU FQOKPCPV UGNGEVKQP OCTMÿ GTU KP H. polymorpha (/C[GT GV CN. ú999; 5QJP GV CN. ú999C). /CTMGT IGPGU WUGF HQT E. coli c P. pastoris UJWVVNG XGEVQTU KPENWFG HIS4 CPF ARG4 HTQO GKVJGT S. cerevisiae QT P. pastoris CPF P. pastoris ADE1 CPF URA3. þWTVJGTOQTG IGPGU VJCV EQPHGT TGUKUVCPEG VQ VJG FTWIU DNCUVKEKFKP, \GQEKP, CPF ÷4úü CTG PQYCFC[U WUGF HQT OQNGEWNCT IGPGVKE OCPKRWNCVKQPU QH P. pastoris (%GTGIJKPQ GV CN. 2ýýú; /WMCKÿ [COC GV CN. 2ýý2; 5EQTGT GV CN. ú994). ûTQRCICVKQP QH RNCUOKFU KP H. polymorpha CPF P. pastoris KU GUVCDNKUJGF D[ CWVQPQOQWU TGRNKECVKPI UGSWGPEGU (#ò5•U). 6JGUG &0# HTCIOGPVU CTG EWTTGPVN[ WUGF KP XCTKQWU UJWVVNG XGEVQTU (%TGII GV CN. ú9üó; òQIIGPMCOR GV CN. ú9üô; 5QJP GV CN. ú99ô; 5QJP GV CN. ú999D). ÷GPGTCNN[, UJWVVNG XGEVQTU VJCV EQPVCKP CP #ò5 CTG PQV OCKPVCKPGF WPFGT PQPÿUGNGEVKXG EQPFKVKQPU. 6Q QDVCKP UVCDNG VTCPUHQTOCPVU, VJGUG XGEVQTU ECP DG KPVGITCVGF KPVQ VJG IGPQOG QH DQVJ [GCUVU. +P H. polymorpha, VJKU ECP DG CEJKGXGF XKC NGIKVKOCVG QT KNNGIKVKOCVG TGEQODKPCVKQP. 6CTIGVGF KPVGITCÿ VKQP KU TQWVKPGN[ CEJKGXGF WUKPI NKPGCTK\GF RNCUOKFU, YJKEJ TGUWNVU KP CFFKVKXG KPVGÿ ITCVKQP QT IGPG TGRNCEGOGPV FGRGPFKPI QP C UKPING QT FQWDNG ETQUUÿQXGT TGEQODKÿ PCVKQP GXGPV, TGURGEVKXGN[. 6Q FCVG, XCTKQWU YC[U GZKUV VQ UGNGEV HQT UKPING QT OWNVKRNG KPVGITCPVU. 7.2.2 ©ZRTGUUKQP ENQPKPI UVTCVGIKGU 6CMKPI CFXCPVCIG QH VJG HCEV VJCV VJG S. cerevisiae LEU2 IGPG FQGU PQV HWNN[ HWPEÿ VKQPCNN[ EQORNGOGPV VJG H. polymorpha leu1.1 UVTCKP, UKPINGÿEQR[ KPVGITCPVU YKNN HQTO UOCNN EQNQPKGU YJGTGCU OWNVKEQR[ KPVGITCPVU YKNN HQTO NCTIG EQNQPKGU QP UGÿ NGEVKXG OGFKC. # UKOKNCT UVTCVGI[ ECP DG WUGF YJGP VJG H. polymorpha ura3 UVTCKP
2úý
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
KU VTCPUHQTOGF YKVJ C NKPGCTK\GF XGEVQT EQPVCKPKPI VJG S. cerevisiae URA3 IGPG. òGEQODKPCPV UVTCKPU EQWNF DG IGPGTCVGF EQPVCKPKPI WR VQ úýý EQRKGU QH VJG XGEVQT KPVGITCVGF KPVQ VJG IGPQOG (÷CV\MG GV CN. ú99ó). /WNVKEQR[ KPVGITCPVU ECP CNUQ DG QDVCKPGF WUKPI VJG DCEVGTKCN COKPQIN[EQUKFG 3ÿRJQURJQVTCPUHGTCUG (APH) IGPG, YJKEJ EQPHGTU TGUKUVCPEG VQ ÷4úü WPFGT EQPVTQN QH VJG H. polymorpha IN[EGTCNFGÿ J[FGÿ3ÿRJQURJCVG FGJ[FTQIGPCUG (GAPDH) RTQOQVGT. *GTG, EQR[ PWODGTU EQWNF DG EQPVTQNNGF WR VQ óý EQRKGU KPVGITCVGF WUKPI FKHHGTGPV EQPEGPVTCVKQPU QH ÷4úü CPF XCTKQWU FGTKXCVKXGU QH VJG GAPDH RTQOQVGT (5QJP GV CN. ú999C). +P P. pastoris, UVCDNG GZRTGUUKQP UVTCKPU CTG WUWCNN[ QDVCKPGF D[ VCTIGVGF KPVGITCÿ VKQP KPVQ VJG HIS4 OCTMGT IGPG QT VJG CNEQJQN QZKFCUG ú (AOX1) RTQOQVGT. +P UQOG ECUGU, VJG RTQFWEVKQP QH C JGVGTQNQIQWU RTQVGKP KU JKIJGT YJGP P. pastoris UVTCKPU JCXG C /WVU (OGVJCPQN WVKNKUCVKQP UNQY) RJGPQV[RG (%GTGIJKPQ CPF %TGII 2ýýý; *KIIKPU CPF %TGII ú99ü). 5WEJ C UVTCKP ECP DG QDVCKPGF XKC C FQWDNG ETQUUÿQXGT GXGPV KP YJKEJ VJG AOX1 IGPG KU TGRNCEGF KP VQVCN HQT VJG GZRTGUUKQP ECUUGVVG. 6JG TGUWNVKPI UVTCKP JCU C /WVU RJGPQV[RG DGECWUG KV KU HQTEGF VQ TGN[ QP VJG VTCPUETKRÿ VKQPCNN[ YGCMGT AOX2 IGPG HQT OGVJCPQN OGVCDQNKUO. 6Q QDVCKP OWNVKEQR[ GZÿ RTGUUKQP UVTCKPU QH P. pastoris, VJTGG TGNKCDNG OGVJQFU JCXG DGGP FGXGNQRGF. þKTUVN[, D[ WUKPI CP GZRTGUUKQP XGEVQT YJKEJ EQPVCKPU OWNVKRNG JGCFÿVQÿVCKN EQRKGU QH VJG GZRTGUUKQP ECUUGVVG. 5GEQPFN[, CPF SWKVG UKOKNCT VQ VJG H. polymorpha U[Uÿ VGO, WUKPI XGEVQTU VJCV EQPVCKP VJG DCEVGTKCN MCPCO[EKP TGUKUVCPEG IGPG, YJKEJ EQPHGTU TGUKUVCPEG VQYCTFU ÷4úü. +P VJGUG ECUGU, VJG EQR[ PWODGT ECP TQWIJN[ DG EQTTGNCVGF VQ VJG NGXGN QH ÷4úü TGUKUVCPEG. 7UKPI VJKU CRRTQCEJ, UVTCKPU YKVJ 3ý EQRKGU QH VJG GZRTGUUKQP ECUUGVVG EQWNF DG KUQNCVGF (%NCTG GV CN. ú99ú). þKPCNN[, OWNVKEQR[ UVTCKPU ECP DG QDVCKPGF WUKPI VJG \GQEKP TGUKUVCPEG OCTMGT CU C EQORQÿ PGPV QH VJG GZRTGUUKQP XGEVQT. $[ RNCVKPI QP KPETGCUKPI EQPEGPVTCVKQPU QH \GQEKP, VJG OWNVKEQR[ UVTCKPU ECP DG GPTKEJGF HTQO C RQRWNCVKQP QH VTCPUHQTOCPVU. 7.2.3 ÿTQOQVGT U[UVGOU HQT RTQVGKP RTQFWEVKQP ûTQFWEVKQP U[UVGOU DCUGF QP H. polymorpha TGN[ QP VJG WUG QH XCTKQWU RTQOQVGT GNGOGPVU, DQVJ KPFWEKDNG CPF EQPUVKVWVKXG. %QOOQPN[ WUGF RTQOQVGT GNGOGPVU CTG VJQUG FGTKXGF HTQO IGPGU QH VJG OGVJCPQN OGVCDQNKUO RCVJYC[, YJKEJ CTG UVTQPIN[ KPFWEKDNG (6CDNG ú). 6JTGG QH VJGUG, VJG CNEQJQN QZKFCUG RTQOQVGT (ûAOX), FKJ[ÿ FTQZ[CEGVQPG U[PVJCUG RTQOQVGT (ûDHAS), CPF VJG RTQOQVGT QH VJG HQTOCVG FGJ[ÿ FTQIGPCUG (ûFMD) IGPG CTG HWNN[ TGRTGUUGF D[ GZEGUU INWEQUG CPF CTG UVTQPIN[ KPÿ FWEGF D[ OGVJCPQN. >GRTGUUKQP QH VJGUG RTQOQVGTU CNUQ QEEWTU WPFGT INWEQUGÿ QT IN[EGTQNÿNKOKVKPI EQPFKVKQPU (G.I. KP ECTDQPÿNKOKVGF EJGOQUVCV EWNVWTGU). 6JWU, DCÿ UKECNN[, OGVJCPQN KU PQV PGEGUUCT[ HQT RTQVGKP RTQFWEVKQP FTKXGP D[ VJGUG RTQOQVGT GNGOGPVU. 1PG GZCORNG QH C RTQFWEVKQP RTQEGUU KP VJG CDUGPEG QH OGVJCPQN KU VJG JKIJ [KGNF RTQFWEVKQP QH VJG CPVKEQCIWNCPV RGRVKFG JKTWFKP (ñG[FGOCPP GV CN. ú99ó).
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
2úú
6CDNG 1. 2TQOQVGTU WUGF HQT VJG GZRTGUUKQP QH JGVGTQNQIQWU IGPGU KP OGVJ[NQVTQRJKE [GCUVU 2TQOQVGT )GPG QH QTKIKP Hansenula polymorpha ûAOX AOX DHAS ûDHAS ûFLD
FLD
ûFMD ûAMO
FMD AMO
ûYNT1 ûYNR1 ûYNI1 ûPHO1 ûPMA1
YNT1 YNR1 YNI1 PHO1 PMA1
ûTEF1/TEF2
TEF1/TEF2
Pichia pastoris ûAOX1 ûFLD1
AOX1 FLD1
ûPEX8 ûGAP
PEX8 GAP
ûYPT1
YPT1
2TQVGKP HWPEVKQP
4GHGTGPEG
#NEQJQN QZKFCUG &KJ[FTQZ[CEGVQPG U[Pÿ VJCUG þQTOCNFGJ[FG FGJ[FTQÿ IGPCUG þQTOCVG FGJ[FTQIGPCUG #OKPG QZKFCUG
LGFGDQGT GV CN. ú9üó ðCPQYKE\ GV CN. ú9üó
0KVTCVG VTCPURQTVGT 0KVTCVG TGFWEVCUG 0KVTKVG TGFWEVCUG ûJQURJCVCUG ûNCUOC OGODTCPG *ïÿ #6ûCUG 6TCPUNCVKQP GNQPICVKQP #NEQJQN QZKFCUG þQTOCNFGJ[FG FGJ[FTQÿ IGPCUG ûGTQZKUQOG DKQIGPGUKU ÷N[EGTCNFGJ[FGÿ3ÿ RJQURJCVG FGJ[FTQIGPCUG ÷6ûCUG KPXQNXGF KP UGETGÿ VKQP
$CGTGPFU GV CN. 2ýý2 ÷GNNKUUGP GV CN. ú99ú $TWKPGPDGTI GV CN. ú9ü9 ûGTG\ GV CN. ú99ö $TKVQ GV CN. ú99ô $TKVQ GV CN. ú99ô ûJQPIFCTC GV CN. ú99ü %QZ GV CN. 2ýýý ð#-ñ -KGN, WPRWDÿ NKUJGF %TGII GV CN. ú9ü9 5JGP GV CN. ú99ü LKW GV CN. ú99ó ñCVGTJCO GV CN. ú99ö 5GCTU GV CN. ú99ü
#PQVJGT KPFWEKDNG RTQOQVGT, YJKEJ ECP DG WUGF HQT RTQFWEVKQP RWTRQUGU, KU VJG H. polymorpha COKPG QZKFCUG RTQOQVGT (ûAMO). #OKPG QZKFCUG (#/1) KU VJG MG[ GP\[OG KP VJG OGVCDQNKUO QH RTKOCT[ COKPGU D[ H. polymorpha. 6JG ûAMO KU TGÿ RTGUUGF D[ COOQPKWO CPF KPFWEGF D[ RTKOCT[ COKPGU VQ C NGXGN, YJKEJ KU WR VQ 2ýõ EQORCTGF VQ VJG ûAOX KPFWEGF D[ OGVJCPQN. &WTKPI TGEGPV [GCTU, QVJGT KPFWEÿ KDNG RTQOQVGT GNGOGPVU JCXG DGGP KFGPVKHKGF KP H. polymorpha. 6JGUG KPENWFG RTQOQVGT GNGOGPVU FGTKXGF HTQO IGPGU QH VJG PKVTCVG OGVCDQNKUO, YNT1, YNR1 CPF YNI1 (#XKNC GV CN. ú99ó; $TKVQ GV CN. ú99ô; ûGTG\ GV CN. ú99ö), CPF VJG TGRTGUUKDNG CEKF RJQURJCVCUG (PHO1) RTQOQVGT (ûJQPIFCTC GV CN. ú99ü). 6JGUG RTQOQVGTU EQWNF DG WUGF CU PGY EQPVTQN GNGOGPVU HQT RTQVGKP RTQFWEVKQP KP H. polymorpha. +V OC[ CNUQ DG CFXCPVCIGQWU VQ EQPUVKVWVKXGN[ GZRTGUU VJG FGUKTGF IGPG RTQFWEV. òGÿ EGPVN[, UWEJ RTQOQVGTU JCXG DGGP KFGPVKHKGF KP H. polymorpha G.I. VJG RTQOQVGT QH VJG RNCUOC OGODTCPG *ïÿ#6ûCUG (ûPMA1) (%QZ GV CN. 2ýýý) CPF VJG RTQOQVGTU QH VJG IGPGU GPEQFKPI VJG VTCPUNCVKQP GNQPICVKQP HCEVQTÿú½ (TEF1øTEF2; ð.#.-.ñ. -KGN, WPRWDNKUJGF TGUWNVU). +P P. pastoris, JGVGTQNQIQWU IGPGU CTG IGPGTCNN[ GZRTGUUGF WPFGT EQPVTQN QH VJG ûAOX1, YJKEJ KU VKIJVN[ TGIWNCVGF (6CDNG ú). 6JG ûAOX1 GZRTGUUKQP KU HWNN[ TGRTGUUGF D[ ECTDQP UQWTEGU NKMG INWEQUG CPF IN[EGTQN DWV KU OCZKOCNN[ KPFWEGF FWTKPI
2ú2
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
ITQYVJ QH EGNNU QP OGVJCPQN (6UEJQRR GV CN. ú9üö). # V[RKECN CFXCPVCIG QH VJKU TGIWNCVKQP KU VJCV UVTCKPU GZRTGUUKPI JGVGTQNQIQWU IGPGU, YJQUG RTQFWEVU CTG VQZKE HQT VJG EGNN, ECP DG OCKPVCKPGF D[ ITQYKPI VJKU QTICPKUO WPFGT TGRTGUUKPI EQPFKÿ VKQPU. +P UQOG ECUGU, JQYGXGT, VJG WUG QH OGVJCPQN VQ KPFWEG VJG RTQFWEVKQP QH HQTGKIP RTQVGKPU KU KPEQPXGPKGPV QT KPCRRTQRTKCVG. þQT GZCORNG, CNVJQWIJ PGEGUUCT[ HQT VJG HWNN KPFWEVKQP QH VJG ûAOX1, OGVJCPQN ECP DG C RQVGPVKCN HKTGÿJC\CTF. &WTKPI VJG TGEGPV RCUV, CNVGTPCVKXG RTQOQVGT GNGOGPVU JCXG DGEQOG CXCKNCDNG HQT VJG RTQÿ FWEVKQP QH JGVGTQNQIQWU RTQVGKPU D[ P. pastoris. # UGEQPF TGIWNCVGF RTQOQVGT GNGÿ OGPV QTKIKPCVGU HTQO VJG INWVJCVJKQPGÿFGRGPFGPV HQTOCNFGJ[FG FGJ[FTQIGPCUG ú (FLD1) IGPG (5JGP GV CN. ú99ü). 6JG GPEQFGF RTQVGKP KU C MG[ GP\[OG TGSWKTGF HQT VJG OGVCDQNKUO QH OGVJ[NCVGF COKPGU CPF OGVJCPQN. 6JG ûFLD1 KU KPFWEGF D[ DQVJ OGVJCPQN CPF OGVJ[NCOKPG CPF GZRTGUUKQP NGXGNU CTG EQORCTCDNG VQ VJQUG QDVCKPGF YKVJ OGVJCPQN KPFWEGF ûAOX1. #PQVJGT UVTQPI RTQOQVGT WUGF HQT RTQVGKP RTQFWEVKQP KP P. pastoris KU VJG IN[EGTCNFGJ[FGÿ3ÿRJQURJCVG FGJ[FTQIGPCUG RTQOQVGT (ûGAP) (ñCVGTJCO GV CN. ú99ö). 6JG ûGAP YCU UJQYP VQ FTKXG C EQPUVKVWVKXG, JKIJÿNGXGN GZRTGUUKQP QP INWEQUG, IN[EGTQN, CPF OGVJCPQN. # V[RKECN GZCORNG HQT VJG UWEEGUUÿ HWN WUG QH VJKU CNVGTPCVKXG RTQOQVGT KU VJG RTQFWEVKQP CPF CUUGODN[ QH JGRCVKVKU $ UWTHCEG CPVKIGP RCTVKENGU (8CUUKNGXC GV CN. 2ýýú). *QYGXGT, HQT UQOG CRRNKECVKQPU VJG JKIJ GZRTGUUKQP NGXGNU QDVCKPGF WUKPI VJG ûAOX1, ûFLD1, QT ûGAP, OC[ PQV DG UWKVCDNG. 6JG JKIJ NGXGNU QH GZRTGUUKQP QDVCKPGF KP VJGUG ECUGU OC[ QXGTYJGNO VJG RQUVÿVTCPUNCVKQPCN OCEJKPGT[ KP VJG EGNN, YJKEJ EQWNF TGUWNV KP OKUHQNFKPI CPFøQT OKUNQECNKUCVKQP. 6Q EKTEWOXGPV UWEJ RTQDNGOU, VYQ OQFGTCVGN[ GZRTGUUKPI RTQOQVGT GNGOGPVU CTG PQYCFC[U CXCKNCDNG HQT VJG P. pastoris RTQFWEVKQP U[UVGO. 1PG QH VJGUG KU VJG PEX8 RTQOQVGT. PEX8 GPEQFGU C RTQVGKP VJCV KU GUUGPVKCN HQT RGTQZKUQOG DKQIGPGÿ UKU (LKW GV CN. ú99ó). 6JG ûPEX8 KU GZRTGUUGF CV C NQY NGXGN QP INWEQUG CPF KU OQFÿ GTCVGN[ KPFWEGF QP OGVJCPQN. # UGEQPF OQFGTCVG RTQOQVGT KU FGTKXGF HTQO VJG YPT1 IGPG, YJKEJ EQFGU HQT C ÷6ûCUG KPXQNXGF KP UGETGVKQP. ûYPT1 OCKPVCKPU NQY EQPUVKVWVKXG GZRTGUUKQP NGXGNU KP OGFKC EQPVCKPKPI GKVJGT QH VJG HQNNQYKPI: INWEQUG, OGVJCPQN, QT OCPPKVQN (5GCTU GV CN. ú99ü).
7.3 5QTVKPI QH JGVGTQNQIQWU RTQVGKPU VQ URGEKHKE UWDEGNNWNCT NQECVKQPU 6JG OQNGEWNCT VQQNDQZGU HQT DQVJ H. polymorpha CPF P. pastoris, FGVCKNGF CDQXG, CTG PGEGUUCT[ HQT VJG KPVTQFWEVKQP CPF GZRTGUUKQP QH HQTGKIP IGPGU KP VJGUG QTICPÿ KUOU. +P VJGUG GWMCT[QVGU, VJG RTQFWEGF RTQVGKPU ECP DG UQTVGF VQ URGEKHKE UWDEGNNWÿ NCT EQORCTVOGPVU, YJKEJ KP UQOG ECUG KU CFXCPVCIGQWU HQT QRVKOCN TGUWNVU. +P QTFGT VQ OGFKCVG URGEKHKE UQTVKPI, VCTIGVKPI UKIPCNU HQT XCTKQWU EGNNWNCT EQORCTVOGPVU JCXG DGGP KFGPVKHKGF. 5KIPCNU, YJKEJ CTG CXCKNCDNG HQT WUG KP DQVJ URGEKGU, KPENWFG UKIPCNU HQT GHHKEKGPV UQTVKPI VQ RGTQZKUQOGU (DQVJ RGTQZKUQOCN OCVTKZ CU YGNN CU VJG RGTQZKUQOCN OGODTCPG), GPFQRNCUOKE TGVKEWNWO (©ò), VJG UGETGVQT[ RCVJYC[, CPF XCEWQNGU. ûTQFWEVKQP CPF UQTVKPI QH JGVGTQNQIQWU RTQVGKPU VQ RGTQZKUQOGU KU FGÿ VCKNGF DGNQY.
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
2ú3
7.3.1 ÿTQVGKP UQTVKPI VQ RGTQZKUQOGU 7.3.1.1 Genes involved in peroxisome biogenesis and degradation &WTKPI VJG NCUV ú2 [GCTU, QWT MPQYNGFIG QP VJG OQNGEWNCT OGEJCPKUOU QH RGTQZÿ KUQOG DKQIGPGUKU CPF FGITCFCVKQP JCU UKIPKHKECPVN[ KPETGCUGF. /GVJ[NQVTQRJKE [GCUVU VWTPGF QWV VQ DG HCXQWTCDNG OQFGN QTICPKUOU HQT RGTQZKUQOG TGUGCTEJ (þCÿ DGT GV CN. ú99ü; 5CMCK CPF 5WDTCOCPK 2ýýý; 8CP FGT -NGK CPF 8GGPJWKU 2ýý2; 8GGPJWKU CPF 8CP FGT -NGK 2ýý2). +P VJGUG QTICPKUOU, RGTQZKUQOG HQTOCVKQP CPF FGITCFCVKQP ECP TGCFKN[ DG OCPKRWNCVGF D[ EJCPIKPI VJG ITQYVJ OGFKWO. ûGTQZÿ KUQOG CDWPFCPEG KU JKIJGUV KP EGNNU VJCV CTG ITQYP QP OGFKC EQPVCKPKPI OGVJCPQN CU UQNG UQWTEG QH ECTDQP CPF GPGTI[. +P VJGUG EGNNU, VJG RGTQZKUQOGU EQPVCKP GPÿ \[OGU VJCV ECVCN[UG VJG HKTUV UVGRU KP OGVJCPQN OGVCDQNKUO, PCOGN[ #1, &*#5, CPF %#6. +P P. pastoris, RGTQZKUQOG RTQNKHGTCVKQP KU CNUQ KPFWEGF FWTKPI ITQYVJ QP QNGKE CEKF. 7PFGT VJGUG EQPFKVKQPU, VJG QTICPGNNGU JCTDQWT GP\[OGU KPXQNXGF KP HCVV[ CEKF OGVCDQNKUO (!ÿQZKFCVKQP RCVJYC[). H. polymorpha EGNNU CTG WPCDNG VQ ITQY QP QNGKE CEKF. ñJGP OGVJCPQNÿITQYP [GCUV EGNNU CTG VTCPUHGTTGF VQ OGFKC KP YJKEJ VJG QTICPÿ GNNGU DGEQOG TGFWPFCPV HQT ITQYVJ (G.I. INWEQUGÿ QT GVJCPQNÿEQPVCKPKPI OGFKC) RGTQZKUQOGU CTG TCRKFN[ FGITCFGF. +P OGVJ[NQVTQRJKE [GCUV, VYQ OQFGU QH RGTQZÿ KUQOG FGITCFCVKQP (CNUQ FGUKIPCVGF RGZQRJCI[) JCXG DGGP FGUETKDGF, PCOGN[ OCETQRGZQRJCI[ CPF OKETQRGZQRJCI[ (5CMCK CPF 5WDTCOCPK 2ýýý; -KO CPF -NKQPUM[ 2ýýý; $GNNW CPF -KGN 2ýý3). +P H. polymorpha, OCETQRGZQRJCI[ KU KPFWEGF WRQP GZRQUWTG QH EGNNU VQ GZEGUU INWEQUG QT GVJCPQN VQ OGVJCPQNÿITQYP EGNNU, YJGTGCU KP P. pastoris, OCETQRGZQÿ RJCI[ KU QPN[ KPFWEGF D[ GVJCPQN. /CETQRGZQRJCI[ KPXQNXGU VJTGG OQTRJQNQIKÿ ECNN[ FKUVKPEV UVGRU: K) UGSWGUVTCVKQP QH KPFKXKFWCN QTICPGNNGU D[ UGXGTCN OGODTCPG NC[GTU, KK) HWUKQP QH VJG UGSWGUVGTKPI OGODTCPG YKVJ VJG XCEWQNG CPF KKK) FGITCFCÿ VKQP QH VJG QTICPGNNCT EQPVGPVU D[ XCEWQNCT J[FTQNCUGU. /KETQRGZQRJCI[ JCU DGGP QDUGTXGF WRQP CFFKVKQP QH INWEQUG VQ OGVJCPQNÿITQYP P. pastoris EGNNU (6WVVNG CPF &WPP ú99ó). 6JKU RTQEGUU KU EJCTCEVGTKUGF D[ VJG GPIWNHOGPV CPF UWDUGSWGPV WRÿ VCMG QH ENWUVGTU QH RGTQZKUQOGU KP VJG XCEWQNG CRRCTGPVN[ YKVJQWV RTKQT UGSWGUVTCÿ VKQP QH VJG QTICPGNNGU. ÷GPGU KPXQNXGF KP RGTQZKUQOG DKQIGPGUKU (PEX IGPGU) JCXG DGGP ENQPGF WUKPI OWVCPVU VJCV NCEM HWPEVKQPCN RGTQZKUQOGU (pex OWVCPVU). H. polymorpha CPF P. pastoris pex OWVCPVU JCXG NQUV VJGKT ECRCEKV[ VQ ITQY QP OGVJCPQN (H. polymorpha, P. pastoris) QT QNGKE CEKF (P. pastoris), YJKEJ CNNQYU VQ ENQPG VJG EQTTGURQPFÿ KPI IGPGU D[ HWPEVKQPCN EQORNGOGPVCVKQP WRQP VTCPUHQTOCVKQP QH VJG UVTCKPU WUKPI IGPG NKDTCTKGU. H. polymorpha pex OWVCPVU YGTG HKTUV FGUETKDGF D[ %TGII GV CN. (ú99ý). 6JGUG OWVCPVU YGTG UGNGEVGF HTQO OGVJCPQN WVKNKUCVKQP FGHKEKGPV UVTCKPU (/WVÿ) VJCV YGTG QDVCKPGF WRQP EJGOKECN OWVCIGPGUKU. /QTG TGEGPVN[, CP CNVGTPCÿ VKXG CRRTQCEJ YCU FGXGNQRGF DCUGF QP IGPG VCIIKPI D[ 4#PFQO KPVGITCVKQP QH LKPGCT &0# (TCIOGPVU (ò#Lþ; 8CP &KjM GV CN. 2ýýú). /QTGQXGT, H. polymorpha QTVJQNQIWGU QH PEX IGPGU VJCV YGTG HKTUV KFGPVKHKGF KP QVJGT QTICPKUOU JCXG DGGP ENQPGF D[ û%ò CRRTQCEJGU WUKPI RTKOGTU DCUGF QP EQPUGTXGF TGIKQPU KP VJGUG IGPGU. 6JGUG CRRTQCEJGU JCXG TGUWNVGF KP VJG ENQPKPI QH ú4 H. polymorpha PEX
2ú4
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
IGPGU KP QWT NCDQTCVQT[ (PEX1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 14, 19, CPF 20). 5KOKÿ NCT CRRTQCEJGU JCXG DGGP WUGF VQ KUQNCVG P. pastoris PEX IGPGU GZEGRV VJCV P. pastoris pex OWVCPVU CTG DQVJ /WVÿ CPF 1NGÿ (FGHGEVKXG VQ ITQY QP QNGKE CEKF; ÷QWNF GV CN ú992; LKW GV CN. ú992). #V RTGUGPV, VJG ENQPKPI QH úó FKHHGTGPV P. pastoris IGPGU JCU DGGP TGRQTVGF (PEX1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 13, 14, 17, 19, CPF 22). þQT C TGEGPV TGXKGY QP VJG HWPEVKQP QH PEX IGPGU, UGG ûWTFWG CPF LC\CTQY (2ýýú). 5GNGEVKQP QH OWVCPVU QH OGVJ[NQVTQRJKE [GCUVU FGHGEVKXG KP RGZQRJCI[ KU DCUGF QP VJG TGVGPVKQP QH RGTQZKUQOCN #1 CEVKXKV[ WRQP CFCRVCVKQP QH OGVJCPQNÿKPFWEGF EGNNU VQ INWEQUG QT GVJCPQN. #1 CEVKXKV[ ECP GCUKN[ DG XKUWCNKUGF D[ CP #1 CEVKXKV[ RNCVG QXGTNC[ CUUC[ (6KVQTGPMQ GV CN. ú99ó). 6JKU CUUC[ KU RGTHQTOGF HQNNQYKPI TGRÿ NKEC RNCVKPI QH OGVJCPQNÿITQYP EQNQPKGU VQ INWEQUGÿEQPVCKPKPI QT GVJCPQNÿ EQPVCKPKPI RNCVGU. þGY JQWTU CHVGT RNCVKPI, #1 CEVKXKV[ KU UKIPKHKECPVN[ TGFWEGF KP ñ6 EGNNU FWG VQ RGTQZKUQOG FGITCFCVKQP, YJGTGCU OWVCPVU FGHGEVKXG KP RGZQRJCI[ TGVCKP VJKU CEVKXKV[. #NVGTPCVKXGN[, VJG UGPUKVKXKV[ QH OWVCPVU VQYCTFU CNN[N CNEQJQN ECP DG WUGF VQ FGVGTOKPG VJG RTGUGPEG QH #1 CEVKXKV[. #NN[N CNEQJQN KU EQPXGTVGF D[ #1 KPVQ CETQNGKP, C JKIJN[ VQZKE EQORQWPF VJCV MKNNU VJG EGNNU. +P OWVCPVU CHÿ HGEVGF KP RGZQRJCI[, #1 CEVKXKV[ YKNN PQV FTQR WRQP CFCRVCVKQP VQ INWEQUG QT GVJCPQN, TGUWNVKPI KP KPETGCUGF UGPUKVKXKV[ VQYCTFU CNN[N CNEQJQN EQORCTGF VQ ñ6 EGNNU (-QOFWWT GV CN. 2ýý2). #V RTGUGPV, C NCTIG PWODGT QH H. polymorpha CPF P. pastoris IGPGU VJCV EQOÿ RNGOGPV OWVCPVU CHHGEVGF KP FKHHGTGPV UVCIGU QH OKETQRGZQRJCI[ CPFøQT OCETQRGZÿ QRJCI[ JCXG DGGP KFGPVKHKGF. +PVGTGUVKPIN[, OCP[ QH VJGUG VWTPGF QWV VQ DG (JQOQÿ NQIWGU QH) MPQYP IGPGU KPXQNXGF KP XCTKQWU RTQVGKP VTCPURQTV RCVJYC[U VQ VJG XCEWQNG (G.I. VPS IGPGU ÿ XCEWQNCT RTQVGKP UQTVKPI, CVT IGPGU ÿ E[VQUQN VQ XCEWQNG VCTIGVKPI, END IGPGU GPFQE[VQUKU QT IGPGU KPXQNXGF KP IGPGTCN CWVQRJCI[). *QYGXGT, HGY IGPGU JCXG DGGP KUQNCVGF VJCV UGGO VQ DG URGEKHKECNN[ KPXQNXGF KP RGZQRJCI[ (C TGEGPV TGXKGY UGG $GNNW CPF -KGN 2ýý3). #NN PEX IGPGU MPQYP VQ FCVG CTG WPKSWG IGPGU CPF QPN[ KPXQNXGF KP RGTQZÿ KUQOG DKQIGPGUKU. /QUV GPEQFG RTQVGKPU (FGUKIPCVGF RGTQZKPU) VJCV HWPEVKQP KP RGTQZKUQOCN OCVTKZ RTQVGKP KORQTV. %GNNU VJCV NCEM VJGUG RGTQZKPU UVKNN U[PVJGUKUG RGTQZKUQOCN OGODTCPGU, DWV CTG JCORGTGF KP OCVTKZ RTQVGKP KORQTV. 1VJGT RGTQZÿ KPU CTG VJQWIJV VQ DG GUUGPVKCN HQT VJG HQTOCVKQP QH VJG RGTQZKUQOCN OGODTCPG (ûGZ3R CPF ûGZú9R). 6JKU XKGY KU DCUGF QP VJG HKPFKPI VJCV EQPUVTWEVGF OWVCPVU KP YJKEJ VJG PEX3 QT PEX19 IGPG KU FKUTWRVGF NCEM RGTQZKUQOCN OGODTCPG TGOÿ PCPVU. 6JG QRRQUKVG RJGPQOGPQP JCU CNUQ DGGP QDUGTXGF. +P H. polymorpha UVTCKPU VJCV UVTQPIN[ QXGTGZRTGUU PEX3, VJG RGTQZKUQOCN OGODTCPG UWTHCEG KU FTCOCVKÿ ECNN[ KPETGCUGF NGCFKPI VQ VJG HQTOCVKQP QH CP GZVTGOGN[ JKIJ PWODGT QH XGT[ UOCNN RGTQZKUQOGU ($CGTGPFU GV CN. ú99ö). 7.3.1.2 Sorting of heterologous proteins to the peroxisomal matrix +P EQPVTCUV VQ OKVQEJQPFTKC CPF EJNQTQRNCUVU, RGTQZKUQOGU FQ PQV EQPVCKP &0# CPF C RTQVGKP U[PVJGUKUKPI OCEJKPGT[. *GPEG, CNN RGTQZKUQOCN RTQVGKPU CTG GPEQFGF D[ PWENGCT IGPGU. ûTGEWTUQTU QH VJGUG RTQVGKPU CTG U[PVJGUKUGF QP HTGG RQN[UQOGU KP VJG E[VQUQN CPF KORQTVGF RQUVÿVTCPUNCVKQPCNN[ KPVQ VJG VCTIGV QTICPGNNG. ûGTQZÿ KUQOCN VCTIGVKPI UKIPCNU (û65) TGUKFG YKVJKP VJG OCVWTG RQN[RGRVKFG. þKTGHN[
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
2úó
NWEKHGTCUG YCU VJG HKTUV RTQVGKP KP YJKEJ C û65 YCU KFGPVKHKGF CU VJG UQÿECNNGF 5-LÿOQVKH: 5-Lÿ%11* (û65ú) (÷QWNF GV CN. ú9üö). LCVGT UVWFKGU TGXGCNGF VJCV VJKU û65ú KU C IGPGTCN CPF SWKVG FGIGPGTCVG UKIPCN KP PCVWTG ($NCVVPGT GV CN. ú992; ©NIGTUOC GV CN. ú99ô; 5QOOGT GV CN. ú992). +P DQVJ OGVJ[NQVTQRJKE [GCUVU, VJG GPÿ \[OGU KPXQNXGF KP OGVJCPQN OGVCDQNKUO, #1, &*#5, CPF %#6, CTG UQTVGF VQ RGTÿ QZKUQOGU XKC VJGKT TGURGEVKXG û65úÿV[RG VCTIGVKPI UKIPCNU (*CPUGP GV CN. ú992). /QWUG FKJ[FTQHQNCVG TGFWEVCUG, DCEVGTKCN ¾ÿNCEVCOCUG (þKI. ú), C HWUKQP RTQVGKP EQPUKUVKPI QH JWOCP KPUWNKPÿNKMG ITQYVJ HCEVQT CPF C ECTTKGT RTQVGKP CPF VJG ÷TGGP þNWQTGUEGPV ûTQVGKP (÷þû) (þKI. 2) CTG GZCORNGU QH JGVGTQNQIQWU RTQVGKPU YJKEJ JCXG DGGP UWEEGUUHWNN[ RTQFWEGF VQ JKIJ NGXGNU CPF UQTVGF VQ VJG RGTQZKUQOCN OCÿ VTKZ, WUKPI VJG û65ú (þCDGT GV CN. ú99ô; *CPUGP GV CN. ú992; ñCVGTJCO GV CN. ú994).
(KI. 1. ©NGEVTQP OKETQITCRJ UJQYKPI C Hansenula polymorpha UVTCKP RTQFWEKPI C û65ú EQPVCKPKPI ¾ÿNCEVCOCUG HWUKQP RTQVGKP. +OOWPQE[VQEJGOKUVT[ WUKPI URGEKHKE CPVKDQFKGU CICKPUV ¾ÿNCEVCOCUG, TGXGCNGF VJCV VJG DCEVGTKCN ¾ÿNCEVCOCUG YCU NQECNK\GF KP RGTQZKUQOGU. M OKVQEJQPFTKQP, N PWENGWU, P RGTQZKUQOG. $CT = ý.ó ËO.
2úô
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
(KI. ÿ. # HNWQTGUEGPEG OKETQUEQR[ KOCIG QH Hansenula polymorpha EGNNU GZRTGUUKPI ÷þûÿ 5-L. ûGTQZKUQOCN NQECNK\CVKQP QH VJG ÷þû KU GXKFGPV
6JG 5-LÿOQVKH KU PQV VJG QPN[ û65 VJCV JCU DGGP KFGPVKHKGF. òCV VJKQNCUG YCU VJG HKTUV RTQVGKP KP YJKEJ CP CNVGTPCVKXG û65 YCU KFGPVKHKGF ECNNGF û652. 6JG û652 EQPUGPUWU UGSWGPEG (òø-)(Lø8ø+)îó(*øí)(Lø#) (÷NQXGT GV CN. ú994) TGUKFGU CV VJG 0ÿVGTOKPWU QH VJG RTQVGKP. 5WDUGSWGPVN[, û652 UGSWGPEGU YGTG KFGPVKHKGF KP UGXÿ GTCN QVJGT RGTQZKUQOCN RTQVGKPU NKMG YCVGTOGNQP OCNCVG FGJ[FTQIGPCUG, [GCUV VJKQÿ NCUGU, CPF H. polymorpha COKPG QZKFCUG (þCDGT GV CN. ú99ó; ÷KGVN GV CN. ú994). 6JKU UKIPCN KU CNUQ UWKVCDNG VQ FKTGEV JGVGTQNQIQWU RTQVGKPU VQ VJG RGTQZKUQOCN OCVTKZ (ñCVGTJCO GV CN. ú994). 6JG CFXCPVCIG QH VJG CEEWOWNCVKQP QH RTQVGKPU KP RGTQZKUQOGU KU QDXKQWU KP ECUGU YJGP VJG RTQVGKPU CTG VQZKE HQT VJG JQUV QTICPKUO. +P VJKU ECUG, VJG RGTQZÿ KUQOCN OGODTCPG HQTOU C DCTTKGT, RTGXGPVKPI VJCV VJG RTQVGKPU ECP GZGTV VJGKT VQZKE CEVKXKV[ VQ VJG [GCUV EGNN. #PQVJGT RQVGPVKCN CFXCPVCIG QH UVQTCIG KP RGTQZKUQOGU KU VJG CDUGPEG QH OQFKH[KPI GP\[OGU KPUKFG VJG QTICPGNNG. 7PFGUKTGF OQFKHKECVKQPU NKMG G.I. IN[EQU[NCVKQP CTG RTGXGPVGF KP VJKU YC[. #NUQ, RTQVGKPU, YJKEJ CTG UGPUKÿ VKXG VQ RTQVGQN[VKE FGITCFCVKQP, CTG RTQVGEVGF HTQO RTQVGQN[UKU KPUKFG VJG OCVTKZ QH VJG QTICPGNNG. +P EQPENWUKQP: KP ECUG QH RTQFWEVKQP QH UQNWDNG JGVGTQNQIQWU RTQÿ VGKPU, VJG RGTQZKUQOCN OCVTKZ KU CP GZEGNNGPV UVQTCIG EQORCTVOGPV VQ GUVCDNKUJ C TGÿ NKCDNG EGNN HCEVQT[. 7.3.1.3 Sorting of heterologous membrane proteins to the peroxisomal membrane 7R VQ PQY, C TGNKCDNG U[UVGO HQT VJG RTQFWEVKQP QH JGVGTQNQIQWU OGODTCPG RTQÿ VGKPU KU PQV CXCKNCDNG. 5WEJ C U[UVGO KU QH WVOQUV KORQTVCPEG HQT XCTKQWU TGCUQPU G.I. VQ HCEKNKVCVG TCVKQPCN FTWI FGUKIP. 6JG PGGF HQT VJKU DGEQOGU ENGCT HTQO VJG QDÿ UGTXCVKQP VJCV OCP[ JWOCP FKUGCUGU CTG ECWUGF D[ OCNHWPEVKQPKPI QH OGODTCPG RTQVGKPU. /QTGQXGT, HWPFCOGPVCN TGUGCTEJ, NKMG TGUQNXKPI VJG UVTWEVWTG CPF HWPEÿ
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
2úö
VKQPCN UVWFKGU QH KORQTVCPV OGODTCPG RTQVGKPU TGNKGU QP VJG CXCKNCDKNKV[ QH TGNCÿ VKXGN[ NCTIG COQWPVU QH DKQNQIKECNN[ CEVKXG OGODTCPG RTQVGKPU. ñG JCXG UJQYP VJCV VJG H. polymorpha RGTQZKUQOCN OGODTCPG OC[ DG VJG PKEJG QH EJQKEG VQ QXGTRTQFWEG OGODTCPG RTQVGKPU. +P VJKU QTICPKUO, DWV PQV KP P. pastoris VJCV PQTÿ OCNN[ ITQYU QP QNGCVG KP EQPVTCUV VQ H. polymorpha, GZEGUUKXG RGTQZKUQOCN OGOÿ DTCPGU FGXGNQRGF VJCV JCTFN[ EQPVCKP RTQVGKP EQORQPGPVU CPF VJWU CTG CP GZEGNNGPV UVQTCIG RNCEG HQT JGVGTQNQIQWU OGODTCPG RTQVGKPU (þKI. 3) (8GGPJWKU GV CN. ú99ý). 5QTVKPI VQ VJGUG OGODTCPGU ECP DG OGFKCVGF D[ VJG VCTIGVKPI UKIPCN QH VJG H. polymorpha RGTQZKUQOCN OGODTCPG RTQVGKP ûGZ3R ($CGTGPFU GV CN. ú99ô). ûTQOKUKPI TGUWNVU HQT VJG QXGTRTQFWEVKQP QH JWOCP %%òó CPF %î%ò4 (*+8ÿú EQÿTGEGRVQTU) JCXG CNTGCF[ DGGP QDVCKPGF (ñCIPGT GV CN. 2ýýú). 6JWU, H. polymorpha JCU RTQXGP VQ DG C XCNWCDNG VQQN HQT VJG JGVGTQNQIQWU RTQFWEVKQP QH OGODTCPG RTQVGKPU. #P CNVGTPCVKXG OQFG VQ GPJCPEG VJG RGTQZKUQOCN OGODTCPG UWTHCEG KU VQ QXGTÿ RTQFWEG *RûGZ3R, YJKEJ NGCFU VQ OCUUKXG RGTQZKUQOG RTQNKHGTCVKQP CPF VJG HQTÿ OCVKQP QH PWOGTQWU UOCNN RGTQZKUQOGU ($CGTGPFU GV CN. ú99ö).
(KI. 3. ©NGEVTQP OKETQITCRJ QH C ñ6 Hansenula polymorpha EGNN KPEWDCVGF KP QNGKE CEKF EQPVCKPKPI OGFKWO. 6JG RTGUGPEG QH PWOGTQWU RGTQZKUQOCN OGODTCPG NC[GTU KU GXKFGPV. M OKVQEJQPFTKQP, N PWENGWU, arrow RGTQZKUQOCN OGODTCPG NC[GTU. $CT = ý.ó ËO.
2úü
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
7.3.2 ÿTQVGKP UGETGVKQP #NVJQWIJ RGTQZKUQOGU ECP CEV UWEEGUUHWNN[ CU C UVQTCIG EQORCTVOGPV HQT JGVGTQNQÿ IQWUN[ RTQFWEGF RTQVGKPU, KP OCP[ ECUGU, UGETGVKQP KU HCXQWTCDNG, G.I. YJGP OQFKÿ HKECVKQP QH VJG RTQFWEGF RTQVGKP KU PGEGUUCT[ HQT CEVKXCVKQP CPFøQT HWPEVKQPKPI (G.I. HQTOCVKQP QH FKÿUWNRJKFG DQPFU, IN[EQU[NCVKQP). 5GETGVKQP CNUQ PGEGUUKVCVGU TGEQXÿ GT[ QH VJG RTQFWEGF RTQVGKP HTQO VJG EWNVWTG HNWKF. 7UKPI H. polymorpha QT P. pastoris, RWTKHKECVKQP QH VJG UGETGVGF RTQVGKP VQ JQOQIGPGKV[ KU TGNCVKXGN[ UKORNG UKPEG EQPFKVKQPU ECP DG ETGCVGF VJCV GPFQIGPQWU RTQVGKP UGETGVKQP KU GZVTGOGN[ NQY. +P VJKU YC[, VJG RTQVGKP VJCV KU RTQFWEGF EQPUVKVWVGU >9ýõ QH VJG GZVTCEGNNWNCT RTQVGKP CPF ECP DG TGCFKN[ TGEQXGTGF. 6TCPUNQECVKQP QH RTQVGKPU KPVQ VJG ©ò KU VJG HKTUV UVGR EQOOKVVGF KP VJG UGETGVQT[ RCVJYC[. /QUV RTQVGKPU VJCV GPVGT VJG ©ò EQPVCKP CP 0ÿVGTOKPCN GZVGPUKQP MPQYP CU UKIPCN RGRVKFG, UKIPCN UGSWGPEG, QT NGCFGT UGSWGPEG. 5KIPCN RGRVKFGU EQPVCKP C VTKRCTVKVG UVTWEVWTG. 6JG HKTUV RCTV EQPUKUVU QH QPG VQ HKXG COKPQ CEKFU, YJKEJ HQTO C RQUKVKXGN[ EJCTIGF TGIKQP. 6JG EGPVTCN RCTV KU C J[FTQRJQDKE EQTG QH UGXGP VQ HKHÿ VGGP COKPQ CEKFU, YJKEJ KU HKPCNN[ HQNNQYGF D[ C RQNCT TGIKQP QH VJTGG VQ UGXGP COKPQ CEKFU. 6JKU NCUV RQNCT TGIKQP KU VJG UKVG HQT TGEQIPKVKQP D[ VJG UKIPCN RGRVKÿ FCUG EQORNGZ (5û%). 0GZV VQ RTQVGKPU EQPVCKPKPI C RTGUGSWGPEG, UQOG UGETGVQT[ RTQVGKPU EQPVCKP C RTGRTQUGSWGPEG G.I. VJG S. cerevisiae OCVKPI HCEVQT ú½ (/þú½) (%CRNCP CPF -WTjCP ú99ú). +P VJGUG ECUGU, VJG RTGUGSWGPEG KU ENGCXGF D[ VJG ©òÿ DQTPG 5û% YJGTGCU VJG RTQUGSWGPEG KU TGOQXGF KP VJG NCVG ÷QNIK D[ VJG GPFQRTQÿ VGCUG -GZ2R. -GZ2R ENGCXGU VJG RTQUGSWGPEG CV VJG FKDCUKE UGSWGPEG (-ò QT òò Á CNVGTPCVKXGN[; L[Uÿ#TI QT #TIÿ#TI) (/K\WPQ GV CN. ú9ü9). 8CTKQWU JGVGTQNQIQWU UKIPCN UGSWGPEGU CTG CXCKNCDNG HQT WUG KP H. polymorpha, KPENWFKPI VJG NGCFGT UGSWGPEGU QH S. cerevisiae /þú½, S. cerevisiae KPXGTVCUG, Schwanniomyces occidentalis INWEQCO[NCUG, CPF VJG NGCFGT UGSWGPEG QH VJG J[ÿ RGTIN[EGOKE JQTOQPG HTQO VJG UJQTG ETCD Carcinus maenas (*QFIMKPU GV CN. ú993; òQFTKIWG\ GV CN. ú99ô; XCP FGT *GKFG GV CN. 2ýý2; 8GCNG GV CN. ú992; ñG[FGÿ OCPP GV CN. ú99ó). +P H. polymorpha, JGVGTQNQIQWU RTQVGKPU ECTT[KPI VJGKT GPFQIGÿ PQWU NGCFGT UGSWGPEGU CNUQ CTG GHHGEVKXG, CU KP VJG ECUG QH JGVGTQNQIQWU RTQFWEVKQP QH Humicola insolens EGNNWNCUG ++ CPF Z[NCPCUG + (/WNNGT GV CN. ú99ü). 1PN[ TGÿ EGPVN[, JQOQNQIQWU H. polymorpha UKIPCN UGSWGPEGU JCXG DGGP KFGPVKHKGF, PCOGN[ VJG NGCFGT UGSWGPEGU QH CEKF RJQURJCVCUG (GPEQFGF D[ PHO1), ECTDQZ[RGRVKFCUG ù (%ûù), CPF $Kû ($GNNW GV CN. ú999). ÷TGGP þNWQTGUEGPV ûTQVGKP (÷þû) KU GHHKEKGPVN[ UQTVGF VQ VJG ©ò WUKPI VJG 0ÿVGTOKPCN 3ý COKPQ CEKFU QH H. polymorpha $Kû (þCDGT GV CN. 2ýý2). *QYGXGT, VJG HWPEVKQPCNKV[ QH VJGUG NGCFGT UGSWGPEGU VQ OGFKÿ CVG UGETGVKQP QH HQTGKIP RTQVGKPU TGOCKPU VQ DG GUVCDNKUJGF. 7UKPI P. pastoris CU C EGNN HCEVQT[, UWEEGUUHWN UGETGVKQP QH JGVGTQNQIQWU RTQVGKPU KU KP OQUV ECUGU QDVCKPGF YJGP UGETGVKQP KU OGFKCVGF D[ VJG S. cerevisiae /þú½. #PQVJGT EQOOQPN[ WUGF UGETGVKQP UKIPCN KU VJG P. pastoris CEKF RJQURJCVCUG (PHO1) UKIPCN (%GTGIJKPQ CPF %TGII 2ýýý). #NVJQWIJ VJG UWEEGUU KP UGETGVKQP QH OCP[ HQTGKIP RTQVGKPU WUKPI VJG /þú½ QT VJG PHO1 UKIPCNU, UQOG RTQVGKPU CTG PQV GHHKEKGPVN[ UGETGVGF QT VJG UGETGVGF RTQVGKP KU KORTQRGTN[ RTQEGUUGF. 6Q FCVG, XCTKÿ QWU QVJGT UKIPCN UGSWGPEGU CTG WUGF VQ GUVCDNKUJ UGETGVKQP D[ P. pastoris. 6JGUG KPÿ ENWFG, PCVKXG UGETGVKQP UKIPCNU RTGUGPV QP VJG JGVGTQNQIQWU RTQVGKP, VJG UKIPCN UGÿ
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
2ú9
SWGPEG QH VJG R÷-L MKNNGT RTQVGKP, VJG UGETGVKQP UKIPCN QH VJG MKNNGT VQZKP QH Pichia acaciae, CPF VJG NGCFGT UGSWGPEG QH Phaseolus vulgaris RJ[VQJGOCIINWVKPKP (-CVQ GV CN. 2ýýú; òCGOCGMGTU GV CN. ú999).
7.4 %QPENWFKPI TGOCTMU 6Q FCVG, C DTQCF TCPIG QH EGNN HCEVQTKGU KU CXCKNCDNG VQ RTQFWEG VJG XCTKQWU RTQVGKPU VJCV CTG WUGF HQT HWPFCOGPVCN TGUGCTEJ, HQT VGEJPKECN GP\[OGU CPF HQT RJCTOCEGWÿ VKECN RWTRQUGU. /GVJ[NQVTQRKE [GCUVU JCXG DGEQOG RTGHGTTGF JQUVU HQT JGVGTQNQIQWU RTQVGKP RTQFWEVKQP FWTKPI VJG NCUV HGY [GCTU. 5GXGTCN GZCORNGU GZKUV KP YJKEJ RTQÿ VGKPU CTG RTQFWEGF YKVJ JKIJGT [KGNFU EQORCTGF VQ QVJGT U[UVGOU. 6JG UCHG CPF EQPXGPKGPV WUG QH OGVJ[NQVTQRJU HQT EQOOGTEKCN RWTRQUGU KU JKIJNKIJVGF D[ VJG RTQFWEVKQP QH C PQXGN JGRCVKVKU $ XCEEKPG D[ H. polymorpha. 6JKU TGEQODKPCPV RTQFWEV JCU RCUUGF ENKPKECN GZCOKPCVKQPU CPF KU PQY QP VJG OCTMGV (÷GNNKUUGP CPF /GNDGT ú99ô). #NUQ, H. polymorpha UGGOU VQ DG VJG QTICPKUO QH EJQKEG HQT VJG RTQFWEVKQP QH IGNÿHQTOKPI RTQVGKPU (G.I. IGNCVKP) DGECWUG KV EQPVCKPU CP GPFQIGÿ PQWU RTQN[N 4ÿJ[FTQZ[NCUG CEVKXKV[, GUUGPVKCN HQT VJG IGNÿHQTOKPI CEVKXKV[ QH VJGUG RTQVGKPU, YJKEJ KU PQV QDUGTXGF KP P. pastoris (FG $TWKP 2ýý2; -GK\GTÿ÷WPPKPM GV CN. 2ýýý; 0QMGNCKPGP GV CN. 2ýýú). #RCTV HTQO EQOOGTEKCNN[ RTQFWEGF RTQVGKPU, OGVJ[NQVTQRJU, CPF KP RCTVKEWNCT H. polymorpha OKIJV DG VJG RTGHGTTGF JQUV HQT VJG U[PVJGUKU QH OGODTCPG RTQVGKPU. 6JG RGTQZKUQOCN OGODTCPGU KPFWEGF FWTKPI KPÿ EWDCVKQP QH EGNNU QP QNGCVG OC[ RTQXKFG C WUGHWN PKEJG HQT VJG CEEWOWNCVKQP QH JGVÿ GTQNQIQWU OGODTCPG RTQVGKPU. þWTVJGT FGXGNQROGPV QH VJKU U[UVGO YQWNF UVKOWÿ NCVG VJG CPCN[UKU QH VJKU KORQTVCPV ENCUU QH RTQVGKPU.
#EMPQYNGFIGOGPVU ñG VJCPM -NCCU 5jQNNGOC HQT JKU CUUKUVCPEG KP RTGRCTKPI VJG RKEVWTGU. /. X.F. * KU UWRRQTVGF D[ C ITCPV QH VJG 0GVJGTNCPFU 6GEJPQNQI[ þQWPFCVKQP (56ñ), YJKEJ KU UWDUKFKUGF D[ VJG 0GVJGTNCPFU 1TICPKUCVKQP HQT 5EKGPVKHKE òGUGCTEJ (0ñ1). +.ð. X.F. -NGK JQNFU C ûKQPKGT ITCPV, UWDUKFKUGF D[ 0ñ1ø#Lñ.
þGHGTGPEGU #ICRJQPQX /1, ûQ\P[CMQXUMK #+, $QIFCPQXC #+, 6GTÿ#XCPGU[CP /& (ú994) +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH VJG LEU2 IGPG QH Hansenula polymorpha. ùGCUV úý:óý9ÿóú3 #XIGTKPQU ÷%, 6WTPGT $÷, ÷QTGNKEM -ð, ûCRGPFKGEM #, ñG[FGOCPP 7, ÷GNNKUUGP ÷ (2ýýú) ûTQFWEVKQP CPF ENKPKECN FGXGNQROGPV QH C Hansenula polymorphaÿFGTKXGF û©÷[NCVGF JKTWFKP. 5GOKP 6JTQOD *GOQUV 2ö:3óöÿ3ö2
22ý
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
#XKNC ð, ûGTG\ /&, $TKVQ 0, ÷QP\CNG\ %, 5KXGTKQ ð/ (ú99ó) %NQPKPI CPF FKUTWRVKQP QH VJG YNR1 IGPG GPEQFKPI VJG PKVTCVG TGFWEVCUG CRQGP\[OG QH VJG [GCUV Hansenula polymorpha. þ©$5 LGVV 3ôô:ú3öÿú42 $CGTGPFU òð5, òCUOWUUGP 5ñ, *KNDTCPFU ò©, XCP FGT *GKFG /, þCDGT -0, òGWXGMCOR û6, -KGN ð#-ñ, %TGII ð/, XCP FGT -NGK +ð, 8GGPJWKU / (ú99ô) 6JG Hansenula polymorpha PER9 IGPG GPEQFGU C RGTQZKUQOCN OGODTCPG RTQVGKP GUUGPVKCN HQT RGTQZKUQOG CUUGODN[ CPF KPVGITKV[. ð $KQN %JGO 2öú:üüüöÿüü94 $CGTGPFU òð5, 5CNQOQPU þ#, -KGN ð#-ñ, XCP FGT -NGK +ð, 8GGPJWKU / (ú99ö) &GXKCPV ûGZ3R NGXGNU CHHGEV PQTOCN RGTQZKUQOG HQTOCVKQP KP Hansenula polymorpha: C UJCTR KPETGCUG QH VJG RTQVGKP NGXGN KPFWEGU VJG RTQNKHGTCVKQP QH PWOGTQWU, UOCNN RTQVGKPÿ KORQTV EQORGVGPV RGTQZKUQOGU. ùGCUV ú3:ú449ÿú4ô3 $CGTGPFU òð5, 5WNVGT ÷ð, ðGHHTKGU 6ñ, %TGII ð/, 8GGPJWKU / (2ýý2) /QNGEWNCT EJCTCEÿ VGTK\CVKQP QH VJG Hansenula poN[OQTRJC FLD1 IGPG GPEQFKPI HQTOCNFGJ[FG FGJ[FTQÿ IGPCUG. ùGCUV ú9:3öÿ42 $GNNW #ò, XCP FGT -NGK +ð, òGEJKPIGT -$, ùCXW\ /, 8GGPJWKU /, -KGN ð#-ñ (ú999) %JCTCEVGTK\CVKQP QH VJG Hansenula polymorpha CPY IGPG GPEQFKPI ECTDQZ[RGRVKFCUG ù. ùGCUV úó:úüúÿúü9 $GNNW #ò, -KGN ð#-ñ (2ýý3) ûGTQZKUQOG FGITCFCVKQP KP [GCUV. /KET òGU 6GEJP KP RTGUU $NCVVPGT ð, 5YKPMGNU $, &QTUCO *, ûTQURGTQ 6, 5WDTCOCPK 5, %NC[VQP % (ú992) ÷N[EQUQOG CUUGODN[ KP VT[RCPQUQOGU: XCTKCVKQPU KP VJG CEEGRVCDNG FGIGPGTCE[ QH C %11*ÿ VGTOKPCN OKETQDQF[ VCTIGVKPI UKIPCN. ð %GNN $KQN úú9:úú29ÿúú3ô $QGFGMGT $÷ (2ýýú) ûTQFWEVKQP RTQEGUUGU QH NKEGPUGF TGEQODKPCPV HCEVQT 8+++ RTGRCTCÿ VKQPU. 5GOKP 6JTQOD *GOQUV 2ö:3üóÿ394 $QIFCPQXC #+, #ICRJQPQX /1, 6GTÿ#XCPGU[CP /& (ú99ó) ûNCUOKF TGQTICPK\CVKQP FWTKPI KPVGITCVKXG VTCPUHQTOCVKQP KP Hansenula polymorpha. ùGCUV úú:343ÿ3ó3 $TKVQ 0, #XKNC ð, ûGTG\ /&, ÷QP\CNG\ %, 5KXGTKQ ð/ (ú99ô) 6JG IGPGU YNI1 CPF YNR1, GPÿ EQFKPI PKVTKVG TGFWEVCUG CPF PKVTCVG TGFWEVCUG TGURGEVKXGN[ KP VJG [GCUV Hansenula polymorpha, CTG ENWUVGTGF CPF EQÿQTFKPCVGN[ TGIWNCVGF. $KQEJGO ð 3úö:ü9ÿ9ó $TWKPGPDGTI û÷, ©XGTU /©, ñCVGTJCO *ò, -WKRGTU ð, #TPDGTI #%, #$ ÷ (ú9ü9) %NQPKPI CPF UGSWGPEKPI QH VJG RGTQZKUQOCN COKPG QZKFCUG IGPG HTQO Hansenula polymorpha. $KQEJKO $KQRJ[U #EVC úýýü:úóöÿúôö %CRNCP 5, -WTjCP ð (ú99ú) òQNG QH CNRJCÿHCEVQT CPF VJG /þ CNRJC ú CNRJCÿHCEVQT RTGEWTUQT KP OCVKPI KP [GCUV. ÷GPGVKEU ú2ö:299ÿ3ýö %GTGIJKPQ ÷ûL, %GTGIJKPQ ðL, 5WPIC #ð, ðQJPUQP /#, LKO /, ÷NGGUQP /#, %TGII ð/ (2ýýú) 0GY UGNGEVCDNG OCTMGTøCWZQVTQRJKE JQUV UVTCKP EQODKPCVKQPU HQT OQNGEWNCT IGÿ PGVKE OCPKRWNCVKQP QH Pichia pastoris. ÷GPG 2ô3:úó9ÿúô9 %GTGIJKPQ ðL, %TGII ð/ (2ýýý) *GVGTQNQIQWU RTQVGKP GZRTGUUKQP KP VJG OGVJ[NQVTQRJKE [GCUV Pichia pastoris. þ©/5 /KETQDKQN òGX 24:4óÿôô %NCTG ðð, òQOCPQU /#, òC[OGPV þ$, òQYGFFGT ð©, 5OKVJ /#, ûC[PG //, 5TGGMTKUJPC -, *GPYQQF %# (ú99ú) ûTQFWEVKQP QH OQWUG GRKFGTOCN ITQYVJ HCEVQT KP [GCUV: JKIJÿ NGXGN UGETGVKQP WUKPI Pichia pastoris UVTCKPU EQPVCKPKPI OWNVKRNG IGPG EQRKGU. ÷GPG úýó:2ýóÿ2ú2 %QZ *, /GCF &, 5WFDGT[ û, ©NCPF ò/, /CPPC\\W +, ©XCPU L (2ýýý) %QPUVKVWVKXG GZRTGUÿ UKQP QH TGEQODKPCPV RTQVGKPU KP VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha WUKPI VJG PMA1 RTQOQVGT. ùGCUV úô:úú9úÿú2ý3 %TGII ð/, $CTTKPIGT -ð, *GUUNGT #ù, /CFFGP -ò (ú9üó) Pichia pastoris CU C JQUV U[UVGO HQT VTCPUHQTOCVKQPU. /QN %GNN $KQN ó:33öôÿ33üó
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
22ú
%TGII ð/, /CFFGP -ò, $CTTKPIGT -ð, 6JKNN ÷û, 5VKNNOCP %# (ú9ü9) þWPEVKQPCN EJCTCEVGTKÿ \CVKQP QH VJG VYQ CNEQJQN QZKFCUG IGPGU HTQO VJG [GCUV Pichia pastoris. /QN %GNN $KQN 9:ú3úôÿú323 %TGII ð/, 8CP FGT -NGK +ð, 5WNVGT ÷ð, 8GGPJWKU /, *CTFGT ñ (ú99ý) ûGTQZKUQOGÿFGHKEKGPV OWVCPVU QH Hansenula polymorpha. ùGCUV ô:üöÿ9ö &G $TWKP ©% (2ýý2) òGEQODKPCPV IGNCVKP CPF EQNNCIGP HTQO OGVJ[NQVTQRJKE [GCUVU. ûJ& VJGUKU ñCIGPKPIGP 7PKXGTUKV[, VJG 0GVJGTNCPFU &QJOGP òð, 5VTCUUGT #ñ, *QPGT %$, *QNNGPDGTI %û (ú99ú) #P GHHKEKGPV VTCPUHQTOCVKQP RTQEGFWTG GPCDNKPI NQPIÿVGTO UVQTCIG QH EQORGVGPV EGNNU QH XCTKQWU [GCUV IGPGTC. ùGCUV ö:ô9úÿô92 ©NIGTUOC ù, 8QU #, XCP &GP $/, XCP òQGTOWPF %ñ, XCP FGT 5û, &KUVGN $, 6CDCM *þ (ú99ô) #PCN[UKU QH VJG ECTDQZ[NÿVGTOKPCN RGTQZKUQOCN VCTIGVKPI UKIPCN ú KP C JQOQNQÿ IQWU EQPVGZV KP Saccharomyces cerevisiae. ð $KQN %JGO 2öú:2ô3öóÿ2ô3ü2 þCDGT -0, *CKOC û, *CTFGT ñ, 8GGPJWKU /, #D ÷ (ú994) *KIJN[ÿGHHKEKGPV GNGEVTQVTCPUÿ HQTOCVKQP QH VJG [GCUV Hansenula polymorpha. %WTT ÷GPGV 2ó:3ýóÿ3úý þCDGT -0, -GK\GTÿ÷WPPKPM +, ûNWKO &, *CTFGT ñ, #D ÷, 8GGPJWKU / (ú99ó) 6JG 0ÿ VGTOKPWU QH COKPG QZKFCUG QH Hansenula polymorpha EQPVCKPU C RGTQZKUQOCN VCTIGVKPI UKIPCN. þ©$5 LGVV 3óö:úúóÿú2ý þCDGT -0, ñGUVTC 5, ñCVGTJCO *ò, -GK\GTÿ÷WPPKPM +, *CTFGT ñ, 8GGPJWKU ÷# (ú99ô) þQTGKIP IGPG GZRTGUUKQP KP Hansenula polymorpha. # U[UVGO HQT VJG U[PVJGUKU QH UOCNN HWPEVKQPCN RGRVKFGU. #RRN /KETQDKQN $KQVGEJPQN 4ó:ö2ÿö9 þCDGT -0, ©NIGTUOC ù, *G[OCP ð#, -QNNGT #, LWGTU ÷*, 0WVVNG[ ñ/, 6GTNGEM[ 5ò, ñGP\GN 6ð, 5WDTCOCPK 5 (ú99ü) 7UG QH Pichia pastoris CU C OQFGN GWMCT[QVKE U[UVGO. ûGTQZKUQOG DKQIGPGUKU. /GVJQFU /QN $KQN. úý3:ú2úÿú4ö. þCDGT -0, *CCP ÷ð, $CGTGPFU òð5, -TCO #/, 8GGPJWKU / (2ýý2) 0QTOCN RGTQZKUQOG FGXGNQROGPV HTQO XGUKENGU KPFWEGF D[ VTWPECVGF Hansenula polymorpha ûGZ3R. ð $KQN %JGO 2öö:úúý2ôÿúúý33 ÷CV\MG ò, ñG[FGOCPP 7, ðCPQYKE\ <#, *QNNGPDGTI %û (ú99ó) 5VCDNG OWNVKEQR[ KPVGITCÿ VKQP QH XGEVQT UGSWGPEGU KP Hansenula polymorpha. #RRN /KETQDKQN $KQVGEJPQN 43:ü44ÿü49 ÷GNNKUUGP ÷, ðCPQYKE\ <#, /GTEMGNDCEJ #, ûKQPVGM /, -GWR û, ñG[FGOCPP 7, *QNNGPÿ DGTI %û, 5VTCUUGT #ñ (ú99ú) *GVGTQNQIQWU IGPG GZRTGUUKQP KP Hansenula polymorpha: GHHKEKGPV UGETGVKQP QH INWEQCO[NCUG. $KQVGEJPQNQI[ (0 ù) 9:29úÿ29ó ÷GNNKUUGP ÷, ñG[FGOCPP 7, 5VTCUUGT #ñ, ûKQPVGM /, ðCPQYKE\ <#, *QNNGPDGTI %û (ú992) ûTQITGUU KP FGXGNQRKPI OGVJ[NQVTQRJKE [GCUVU CU GZRTGUUKQP U[UVGOU. 6TGPFU $KQVGEJÿ PQN úý:4ú3ÿ4úö ÷GNNKUUGP ÷, /GNDGT - (ú99ô) /GVJ[NQVTQRJKE [GCUV Hansenula polymorpha CU RTQFWEVKQP QTICPKUO HQT TGEQODKPCPV RJCTOCEGWVKECNU. #T\PGKOKVVGNHQTUEJWPI 4ô:943ÿ94ü ÷KGVN %, þCDGT -0, XCP FGT -NGK +ð, 8GGPJWKU / (ú994) /WVCVKQPCN CPCN[UKU QH VJG 0ÿ VGTOKPCN VQRQIGPKE UKIPCN QH YCVGTOGNQP IN[QZ[UQOCN OCNCVG FGJ[FTQIGPCUG WUKPI VJG JGVGTQNQIQWU JQUV Hansenula polymorpha . ûTQE 0CVN #ECF 5EK 7 5 # 9ú:3úóúÿ3úóó ÷NQXGT ðò, #PFTGYU &ñ, 5WDTCOCPK 5, òCEJWDKPUMK ò# (ú994) /WVCIGPGUKU QH VJG COKPQ VCTIGVKPI UKIPCN QH Saccharomyces cerevisiae 3ÿMGVQCE[Nÿ%Q# VJKQNCUG TGXGCNU EQPÿ UGTXGF COKPQ CEKFU TGSWKTGF HQT KORQTV KPVQ RGTQZKUQOGU KP XKXQ. ð $KQN %JGO 2ô9:öóóüÿöóô3 ÷QWNF 5ð, -GNNGT ÷ÿ#, 5WDTCOCPK 5 (ú9üö) +FGPVKHKECVKQP QH C RGTQZKUQOCN VCTIGVKPI UKIPCN CV VJG ECTDQZ[ VGTOKPWU QH HKTGHN[ NWEKHGTCUG. ð %GNN $KQN úýó:ü9öÿ9ýó
222
/GKU XCP FGT *GKFG, /CTVGP 8GGPJWKU CPF +FC XCP FGT -NGK
÷QWNF 5ð, /E%QNNWO &, 5RQPI #û, *G[OCP ð#, 5WDTCOCPK 5 (ú992) &GXGNQROGPV QH VJG [GCUV Pichia pastoris CU C OQFGN QTICPKUO HQT C IGPGVKE CPF OQNGEWNCT CPCN[UKU QH RGTÿ QZKUQOG CUUGODN[. ùGCUV ü:ôú3ÿô2ü *CPUGP *, &KFKQP 6, 6JKGOCPP #, 8GGPJWKU /, òQIIGPMCOR ò (ú992) 6CTIGVKPI UGÿ SWGPEGU QH VJG VYQ OCjQT RGTQZKUQOCN RTQVGKPU KP VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha. /QN ÷GP ÷GPGV 23ó:2ô9ÿ2öü *KIIKPU &ò, %TGII ð/ (ú99ü) +PVTQFWEVKQP VQ Pichia pastoris. /GVJQFU /QN $KQN úý3:úÿ úó.:úÿúó *KNN ð, &QPCNF -#, ÷TKHHKVJU &©, &QPCNF ÷ (ú99ú) &/51ÿGPJCPEGF YJQNG EGNN [GCUV VTCPUÿ HQTOCVKQP. 0WENGKE #EKFU òGU ú9:óö9ú *QFIMKPU /, /GCF &, $CNNCPEG &ð, ÷QQFG[ #, 5WFDGT[ û© (ú993) ©ZRTGUUKQP QH VJG INWÿ EQUG QZKFCUG IGPG HTQO Aspergillus niger KP Hansenula polymorpha CPF KVU WUG CU C TGRQTVGT IGPG VQ KUQNCVG TGIWNCVQT[ OWVCVKQPU. ùGCUV 9:ô2óÿô3ó *QNNGPDGTI %û, ÷GNNKUUGP ÷ (ú99ö) ûTQFWEVKQP QH TGEQODKPCPV RTQVGKPU D[ OGVJ[NQVTQRJKE [GCUVU. %WTT 1RKP $KQVGEJPQN ü:óó4ÿóôý ðCPQYKE\ <#, ©EMCTV /ò, &TGYMG %, òQIIGPMCOR ò1, *QNNGPDGTI %û, /CCV ð, LGFGDQGT #/, 8KUUGT %, 8GTTKRU %6 (ú9üó) %NQPKPI CPF EJCTCEVGTK\CVKQP QH VJG IGPG GPÿ EQFKPI VJG OCjQT OGVJCPQN CUUKOKNCVQT[ GP\[OG HTQO VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha. 0WENGKE #EKFU òGU ú3:3ý43ÿ3ýô2 -CVQ 5, +UJKDCUJK /, 6CVUWFC &, 6QMWPCIC *, 6QMWPCIC / (2ýýú) ©HHKEKGPV GZRTGUUKQP, RWÿ TKHKECVKQP CPF EJCTCEVGTK\CVKQP QH OQWUG UCNKXCT[ CNRJCÿCO[NCUG UGETGVGF HTQO OGVJ[ÿ NQVTQRJKE [GCUV Pichia pastoris. ùGCUV úü:ô43ÿôóó -GK\GTÿ÷WPPKPM +, 8WQTGNC #, /[NN[JCTjW ð, ûKJNCjCPKGOK 6, -KXKTKMMQ -+, 8GGPJWKU / (2ýýý) #EEWOWNCVKQP QH RTQRGTN[ HQNFGF JWOCP V[RG +++ RTQEQNNCIGP OQNGEWNGU KP URGÿ EKHKE KPVTCEGNNWNCT OGODTCPQWU EQORCTVOGPVU KP VJG [GCUV Pichia pastoris. /CVTKZ $KQN ú9:29ÿ3ô -KO ð, -NKQPUM[ &ð (2ýýý) #WVQRJCI[, E[VQRNCUOÿVQÿXCEWQNG VCTIGVKPI RCVJYC[, CPF RGZQÿ RJCI[ KP [GCUV CPF OCOOCNKCP EGNNU. #PP òGX $KQEJGO ô9:3ý3ÿ342 -KO /&, òJGG 5-, 5GQ ð* (2ýýú) ©PJCPEGF RTQFWEVKQP QH CPVKEQCIWNCPV JKTWFKP KP TGÿ EQODKPCPV Saccharomyces cerevisiae D[ EJTQOQUQOCN FGNVCÿKPVGITCVKQP. ð $KQVGEJPQN üó:4úÿ4ü -QOFWWT ð#, LGCQ #0, /QPCUV[TUMC +, 8GGPJWKU /, -KGN ð#-ñ (2ýý2) 1NF [GNNQY GPÿ \[OG EQPHGTU TGUKUVCPEG QH Hansenula polymorpha VQYCTFU CNN[N CNEQJQN. %WTT ÷GPGV 4ú:4ýúÿ4ýô LGFGDQGT #/, ©FGPU L, /CCV ð, 8KUUGT %, $QU ðñ, 8GTTKRU %6, ðCPQYKE\ <, ©EMCTV /, òQIIGPMCOR ò, *QNNGPDGTI %û (ú9üó) /QNGEWNCT ENQPKPI CPF EJCTCEVGTK\CVKQP QH C IGPG EQFKPI HQT OGVJCPQN QZKFCUG KP Hansenula polymorpha. 0WENGKE #EKFU òGU ú3:3ýô3ÿ3ýü2 LKW *, 6CP î, 8GGPJWKU /, /E%QNNWO &, %TGII ð/ (ú992) #P GHHKEKGPV UETGGP HQT RGTQZÿ KUQOGÿFGHKEKGPV OWVCPVU QH ûKEJKC RCUVQTKU. ð $CEVGTKQN úö4:4943ÿ49óú LKW *, 6CP î, òWUUGNN -#, 8GGPJWKU /, %TGII ð/ (ú99ó) PER3, C IGPG TGSWKTGF HQT RGTQZÿ KUQOG DKQIGPGUKU KP Pichia pastoris, GPEQFGU C RGTQZKUQOCN OGODTCPG RTQVGKP KPÿ XQNXGF KP RTQVGKP KORQTV. ð $KQN %JGO 2öý:úý94ýÿúý9óú /CVVGU ò (2ýýú) 6JG RTQFWEVKQP QH KORTQXGF VKUUWGÿV[RG RNCUOKPQIGP CEVKXCVQT KP Escherichia coli. 5GOKP 6JTQOD *GOQUV 2ö:32óÿ33ô /C[GT #þ, *GNNOWVJ -, 5EJNKGMGT *, LQRG\ÿ7NKDCTTK ò, 1GTVGN 5, &CJNGOU 7, 5VTCUUGT #ñ, XCP LQQP #û (ú999) #P GZRTGUUKQP U[UVGO OCVWTGU: C JKIJN[ GHHKEKGPV CPF EQUVÿ
ö 6JG OGVJ[NQVTQRJKE [GCUVU Hansenula polymorpha CPF Pichia pastoris
223
GHHGEVKXG RTQEGUU HQT RJ[VCUG RTQFWEVKQP D[ TGEQODKPCPV UVTCKPU QH Hansenula polymorpha. $KQVGEJPQN $KQGPI ô3:3ö3ÿ3üú /GTEMGNDCEJ #, ÷QFGEMG 5, ðCPQYKE\ <#, *QNNGPDGTI %û (ú993) %NQPKPI CPF UGSWGPEKPI QH VJG ura3 NQEWU QH VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha CPF KVU WUG HQT VJG IGPGTCVKQP QH C FGNGVKQP D[ IGPG TGRNCEGOGPV. #RRN /KETQDKQN $KQVGEJPQN 4ý:3ôúÿ 3ô4 /K\WPQ -, 0CMCOWTC 6, 1JUJKOC 6, 6CPCMC 5, /CVUWQ * (ú9ü9) %JCTCEVGTK\CVKQP QH KEX2ÿGPEQFGF GPFQRGRVKFCUG HTQO [GCUV Saccharomyces cerevisiae. $KQEJGO $KQÿ RJ[U òGU %QOOWP úó9:3ýóÿ3úú /WMCK[COC *, 1MW /, $CDC /, 5COK\Q 6, *COOQPF #6, ÷NKEM $5, -CVQ 0, 5CMCK ù (2ýý2) Paz2 CPF ú3 QVJGT PAZ IGPG RTQFWEVU TGIWNCVG XCEWQNCT GPIWNHOGPV QH RGTQZÿ KUQOGU FWTKPI OKETQRGZQRJCI[. ÷GPGU %GNNU ö:öóÿ9ý /WNNGT 5, 5CPFCN 6, -CORÿ*CPUGP û, &CNDQIG * (ú99ü) %QORCTKUQP QH GZRTGUUKQP U[UVGOU KP VJG [GCUVU Saccharomyces cerevisiae, Hansenula polymorpha, Klyveromyces lactis, Schizosaccharomyces pombe CPF Yarrowia lipolytica. %NQPKPI QH VYQ PQXGN RTQOQVGTU HTQO Yarrowia lipolytica. ùGCUV ú4:ú2ôöÿú2ü3 0QMGNCKPGP /, 6W *, 8WQTGNC #, 0QVDQJO *, -KXKTKMMQ -+, /[NN[JCTjW ð (2ýýú) *KIJÿ NGXGN RTQFWEVKQP QH JWOCP V[RG + EQNNCIGP KP VJG [GCUV Pichia pastoris. ùGCUV úü:ö9öÿ üýô ûGTG\ /&, ÷QP\CNG\ %, #XKNC ð, $TKVQ 0, 5KXGTKQ ð/ (ú99ö) 6JG YNT1 IGPG GPEQFKPI VJG PKVTCVG VTCPURQTVGT KP VJG [GCUV Hansenula polymorpha KU ENWUVGTGF YKVJ IGPGU YNI1 CPF YNR1 GPEQFKPI PKVTKVG TGFWEVCUG CPF PKVTCVG TGFWEVCUG, CPF KVU FKUTWRVKQP ECWUGU KPCDKNÿ KV[ VQ ITQY KP PKVTCVG. $KQEJGO ð 32ú:39öÿ4ý3 ûJQPIFCTC #, /GTEMGNDCEJ #, -GWR û, ÷GNNKUUGP ÷, *QNNGPDGTI %û (ú99ü) %NQPKPI CPF EJCTCEVGTK\CVKQP QH VJG IGPG GPEQFKPI C TGRTGUUKDNG CEKF RJQURJCVCUG (PHO1) HTQO VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha. #RRN /KETQDKQN $KQVGEJPQN óý:ööÿü4 ûWTFWG û©, LC\CTQY û$ (2ýýú) ûGTQZKUQOG DKQIGPGUKU. #PPW òGX %GNN &GX $KQN úö:öýúÿ öó2 òCGOCGMGTU òð, FG /WTQ L, ÷CVGJQWUG ð#, þQTFJCOÿ5MGNVQP #û (ú999) þWPEVKQPCN RJ[VQÿ JGOCIINWVKPKP (û*#) CPF Galanthus nivalis CIINWVKPKP (÷0#) GZRTGUUGF KP Pichia pastoris EQTTGEV 0ÿVGTOKPCN RTQEGUUKPI CPF UGETGVKQP QH JGVGTQNQIQWU RTQVGKPU GZÿ RTGUUGF WUKPI VJG û*#ÿ© UKIPCN RGRVKFG. ©WT ð $KQEJGO 2ôó:394ÿ4ý3 òQFTKIWG\ L, 0CTEKCPFK ò©, òQEC *, %TGOCVC ð, /QPVGUKPQU ò, òQFTKIWG\ ©, ÷TKNNQ ð/, /W\KQ 8, *GTTGTC L5, &GNICFQ ð/ (ú99ô) +PXGTVCUG UGETGVKQP KP Hansenula polymorpha WPFGT VJG AOX1 RTQOQVGT HTQO Pichia pastoris. ùGCUV ú2:üúóÿü22 òQIIGPMCOR ò, *CPUGP *, ©EMCTV /, ðCPQYKE\ <#, *QNNGPDGTI %û (ú9üô) 6TCPUHQTOCVKQP QH VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha D[ CWVQPQOQWU TGRNKECVKPI CPF KPÿ VGITCVKQP XGEVQTU. /QN ÷GP ÷GPGV 2ý2:3ý2ÿ3ýü 5CMCK ù, 5WDTCOCPK 5 (2ýýý) ©PXKTQPOGPVCN TGURQPUG QH [GCUV RGTQZKUQOGU. #URGEVU QH QTÿ ICPGNNG CUUGODN[ CPF FGITCFCVKQP. %GNN $KQEJGO $KQRJ[U 32:óúÿôú 5EQTGT %#, %NCTG ðð, /E%QODKG ñò, òQOCPQU /#, 5TGGMTKUJPC - (ú994) òCRKF UGNGEVKQP WUKPI ÷4úü QH JKIJ EQR[ PWODGT VTCPUHQTOCPVU QH Pichia pastoris HQT JKIJÿNGXGN HQTÿ GKIP IGPG GZRTGUUKQP. $KQVGEJPQNQI[ (0 ù ) ú2:úüúÿúü4 5GCTU +$, 1'%QPPQT ð, òQUUCPGUG 1ñ, ÷NKEM $5 (ú99ü) # XGTUCVKNG UGV QH XGEVQTU HQT EQPUVKÿ VWVKXG CPF TGIWNCVGF IGPG GZRTGUUKQP KP Pichia pastoris. ùGCUV ú4:öü3ÿö9ý 5JGP 5, 5WNVGT ÷, ðGHHTKGU 6ñ, %TGII ð/ (ú99ü) # UVTQPI PKVTQIGP UQWTEGÿTGIWNCVGF RTQÿ OQVGT HQT EQPVTQNNGF GZRTGUUKQP QH HQTGKIP IGPGU KP VJG [GCUV Pichia pastoris. ÷GPG 2úô:93ÿúý2
224
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5QJP ð*, %JQK ©5, -CPI *#, òJGG ð5, #ICRJQPQX /1, 6GT #XCPGU[CP /&, òJGG 5(ú999C) # FQOKPCPV UGNGEVKQP U[UVGO FGUKIPGF HQT EQR[ÿPWODGTÿEQPVTQNNGF IGPG KPVGÿ ITCVKQP KP Hansenula polymorpha &Lÿú. #RRN /KETQDKQN $KQVGEJPQN óú:üýýÿüýö 5QJP ð*, %JQK ©5, -CPI *#, òJGG ð5, òJGG 5- (ú999D) # HCOKN[ QH VGNQOGTGÿCUUQEKCVGF CWVQPQOQWUN[ TGRNKECVKPI UGSWGPEGU CPF VJGKT HWPEVKQPU KP VCTIGVGF TGEQODKPCVKQP KP Hansenula polymorpha &Lÿú. ð $CEVGTKQN úüú:úýýóÿúýú3 5QJP ð*, %JQK ©5, -KO %*, #ICRJQPQX /1, 6GTÿ#XCPGU[CP /&, òJGG ð5, òJGG 5(ú99ô) # PQXGN CWVQPQOQWUN[ TGRNKECVKPI UGSWGPEG (#ò5) HQT OWNVKRNG KPVGITCVKQP KP VJG [GCUV Hansenula polymorpha &Lÿú. ð $CEVGTKQN úöü:442ýÿ442ü 5QJP ð*, -CPI *#, òCQ -ð, -KO %*, %JQK ©5, %JWPI $*, òJGG 5- (2ýýú) %WTTGPV UVCVWU QH VJG CPVKEQCIWNCPV JKTWFKP: KVU DKQVGEJPQNQIKECN RTQFWEVKQP CPF ENKPKECN RTCEVKEG. #RRN /KETQDKQN $KQVGEJPQN óö:ôýôÿôú3 5QOOGT ð/, %JGPI íL, -GNNGT ÷#, ñCPI %% (ú992) +P XKXQ KORQTV QH HKTGHN[ NWEKHGTCUG KPVQ VJG IN[EQUQOGU QH Trypanosoma brucei CPF OWVCVKQPCN CPCN[UKU QH VJG %ÿVGTOKPCN VCTIGVKPI UKIPCN. /QN $KQN %GNN 3:ö49ÿöó9 6KVQTGPMQ 8+, -GK\GT +, *CTFGT ñ, 8GGPJWKU / (ú99ó) +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH OWÿ VCPVU KORCKTGF KP VJG UGNGEVKXG FGITCFCVKQP QH RGTQZKUQOGU KP VJG [GCUV Hansenula polymorpha. ð $CEVGTKQN úöö:3óöÿ3ô3 6UEJQRR ðþ, $TWUV ûþ, %TGII ð/, 5VKNNOCP %#, ÷KPIGTCU 6ò (ú9üö) ©ZRTGUUKQP QH VJG NCE< IGPG HTQO VYQ OGVJCPQNÿTGIWNCVGF RTQOQVGTU KP Pichia pastoris. 0WENGKE #EKFU òGU úó:3üó9ÿ3üöô 6WVVNG &L, &WPP ñ# (ú99ó) &KXGTIGPV OQFGU QH CWVQRJCI[ QH RGTQZKUQOGU KP OGVJ[NQVTQÿ RJKE [GCUVU. ©WT ð %GNN $KQN ôý:2ü3ÿ29ý 8CP &KjM ò, þCDGT -0, *COOQPF #6, ÷NKEM $5, 8GGPJWKU /, -KGN ð#-ñ (2ýýú). 6CIÿ IKPI Hansenula polymorpha IGPGU D[ TCPFQO KPVGITCVKQP QH NKPGCT &0# HTCIOGPVU (ò#Lþ). /QN ÷GPGV ÷GPQOKEU 2ôô:ô4ôÿôóô. 8CP FGT *GKFG /, *QNNGPDGTI %û, XCP FGT -NGK +ð, 8GGPJWKU / (2ýý2) 1XGTRTQFWEVKQP QH $Kû PGICVKXGN[ CHHGEVU VJG UGETGVKQP QH Aspergillus niger INWEQUG QZKFCUG D[ VJG [GCUV Hansenula polymorpha. #RRN /KETQDKQN $KQVGEJPQN óü:4üöÿ494 8CP FGT -NGK +ð, 8GGPJWKU / (2ýý2) Hansenula polymorpha: C XGTUCVKNG OQFGN QTICPKUO KP RGTQZKUQOG TGUGCTEJ. +P ÷GNNKUUGP ÷ (GF) Hansenula polymorpha $KQNQI[ CPF #RRNKECÿ VKQPU. ñKNG[ÿ8%* 8GTNCI ÷OD*, ñGKPJGKO, RR öôÿ94 8CP FGT -NGK +ð, 8GGPJWKU / (2ýý2) ûGTQZKUQOGU: HNGZKDNG CPF F[PCOKE QTICPGNNGU. %WTT 1RKP %GNN $KQN ú4:óýýÿóýó. 8CUUKNGXC #, %JWIJ &#, 5YCOKPCVJCP 5, -JCPPC 0 (2ýýú) ©HHGEV QH EQR[ PWODGT QP VJG GZRTGUUKQP NGXGNU QH JGRCVKVKU $ UWTHCEG CPVKIGP KP VJG OGVJ[NQVTQRJKE [GCUV Pichia pastoris. ûTQVGKP ©ZRT ûWTKH 2ú:öúÿüý 8GCNG ò#, ÷KWUGRRKP /L, XCP ©KjM */, 5WFDGT[ û©, 8GTTKRU %6 (ú992) &GXGNQROGPV QH C UVTCKP QH Hansenula polymorpha HQT VJG GHHKEKGPV GZRTGUUKQP QH IWCT CÿICNCEVQUKFCUG. ùGCUV ü:3ôúÿ3ö2 8GGPJWKU /, 8CP &KjMGP ðû, ûKNQP 5#, *CTFGT ñ (ú9öü) &GXGNQROGPV QH ET[UVCNNKPG RGTQZÿ KUQOGU KP OGVJCPQNÿITQYP EGNNU QH VJG [GCUV Hansenula polymorpha CPF KVU TGNCVKQP VQ GPXKTQPOGPVCN EQPFKVKQPU. #TEJ /KETQDKQN úúö:úó3ÿúô3 8GGPJWKU /, *CTFGT ñ (ú9üü) /KETQDQFKGU KP [GCUVU: UVTWEVWTG, HWPEVKQP CPF DKQIGPGUKU. /KETQDKQN 5EK ó:34öÿ3óú 8GGPJWKU /., -TCO #/, -WPCW ñ*, *CTFGT ñ (ú99ý) ©ZEGUUKXG OGODTCPG FGXGNQROGPV HQNNQYKPI GZRQUWTG QH VJG OGVJ[NQVTQRJKE [GCUV Hansenula polymorpha VQ QNGKE CEKFÿ EQPVCKPKPI OGFKC. ùGCUV ô:óúúÿóú9
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8GGPJWKU /, 5CNQOQPU þ#, XCP FGT -NGK +ð (2ýýý) ûGTQZKUQOG DKQIGPGUKU CPF FGITCFCVKQP KP [GCUV: C UVTWEVWTGøHWPEVKQP CPCN[UKU. /KETQUE òGU 6GEJ óú:óü4ÿôýý 8GGPJWKU /, XCP FGT -NGK +ð (2ýý2) ûGTQZKUQOGU: UWTRTKUKPIN[ XGTUCVKNG QTICPGNNGU. $KQÿ EJKO $KQRJ[U #EVC úóóó:44ÿ4ö ñCIPGT ò, -NGKP 8, $CGTGPFU òð5, 8GGPJWKU /., ûCVVWU þ (2ýýú) # PQXGN [GCUV JGVGTQNQÿ IQWU GZRTGUUKQP U[UVGO FGUKIPGF HQT OGODTCPG RTQVGKPU RTQFWEVKQP. ùGCUV úü:2üöÿ2üö ñCVGTJCO *ò, 6KVQTGPMQ 8+, *CKOC û, %TGII ð/, *CTFGT ñ, 8GGPJWKU / (ú994) 6JG Hansenula polymorpha PER1 IGPG KU GUUGPVKCN HQT RGTQZKUQOG DKQIGPGUKU CPF GPEQFGU C RGTQZKUQOCN OCVTKZ RTQVGKP YKVJ DQVJ ECTDQZ[ÿ CPF COKPQÿVGTOKPCN VCTIGVKPI UKIPCNU. ð %GNN $KQN ú2ö:ö3öÿö49 ñCVGTJCO *ò, &KICP /©, -QWV\ ûð, LCKT 58, %TGII ð/ (ú99ö) +UQNCVKQP QH VJG Pichia pastoris IN[EGTCNFGJ[FGÿ3ÿRJQURJCVG FGJ[FTQIGPCUG IGPG CPF TGIWNCVKQP CPF WUG QH KVU RTQOQVGT. ÷GPG úüô:3öÿ44 ñG[FGOCPP 7, -GWR û, ûKQPVGM /, 5VTCUUGT #ñ, 5EJYGFGP ð, ÷GNNKUUGP ÷, ðCPQYKE\ <# (ú99ó) *KIJÿNGXGN UGETGVKQP QH JKTWFKP D[ Hansenula polymorphaÿÿCWVJGPVKE RTQEGUUÿ KPI QH VJTGG FKHHGTGPV RTGRTQJKTWFKPU. #RRN /KETQDKQN $KQVGEJPQN 44:3ööÿ3üó
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#DUVTCEV 6JG [GCUV Yarrowia lipolytica JCU DGGP WUGF HQT KPFWUVTKCN DKQEQPXGTUKQPU UKPEG VJG NCVG ý9ü0•U, OCKPN[ D[ VJG HQQF CPF EJGOKECN KPFWUVTKGU. 6JG OQUV KORQTVCPV RTQEGUU KU RTGUGPVN[ VJG RTQFWEVKQP QH EKVTKE CEKF HTQO TCRGUGGF QKN, DWV Y. lipolytica JCU CNUQ DGGP EQPUKFGTGF HQT XCTKQWU QVJGT CRRNKECVKQPU, TCPIKPI HTQO UKPING EGNN RTQVGKP VQ JGVGTQNQIQWU RTQVGKPU RTQFWEVKQP. 5GXGTCN QH VJGUG RTQEGFWTGU JCXG TGEGKXGF C )4#5 UVCVWU, YJKEJ HCEKNKVCVGU HWTVJGT CRRNKECVKQPU QH VJKU [GCUV KP VJG HQQF CPF RJCTOCEGWVKECN KPFWUVTKGU. 6JKU TGXKGY HQEWUGU QP VJG CESWKUKVKQP QH DCUKE MPQYNGFIG QP Y. lipolytica, YJKEJ KU RJ[UKQNQIKECNN[ CPF VCZQPQOKECNN[ XGT[ FKUVCPV HTQO Saccharomyces cerevisiae. 6JCPMU VQ VJG FGXGNQROGPV QH CFXCPEGF IGPGVKE VQQNU HQT DQVJ ENCUUKECN CRRTQCEJGU CPF TGEQODKPCPV &û# VGEJPQNQI[, UKIPKHKECPV RTQITGUUGU JCXG DGGP CEJKGXGF KP UGXGTCN CURGEVU QH KVU WPWUWCN DKQNQI[. 2CTVKEWNCT CVVGPVKQP JCU DGGP RCKF VQ VJG UVWF[ QH GCTN[ UVGRU QH RTQVGKP UGETGVKQP, RGTQZKUQOG DKQIGPGUKU, ECTDQP OGVCDQNKUO CPF WVKNKUCVKQP QH J[FTQRJQDKE UWDUVTCVGU, OKVQEJQPFTKCN DKQNQI[, OQTRJQIGPGVKE EQPVTQN, CPF RTQVGCG TGIWNCVKQP. #P KPVGTPCVKQPCN GHHQTV TGNC[GF D[ VJG )GPQUEQRG YKNN RGTOKV CEEGUU VQ KVU HWNN IGPQOG UGSWGPEG D[ VJG GPF QH ú003, VJWU HCEKNKVCVKPI HWTVJGT FGXGNQROGPVU QH KVU HWPEVKQPCN IGPGVKEU.
8.1 +PVTQFWEVKQP Yarrowia lipolytica KU CP QDNKICVG CGTQDKE [GCUV, YJKEJ KU TQWVKPGN[ KUQNCVGF HTQO FKHHGTGPV HQQF OGFKC (EJGGUGU, UCWUCIGUãù. 6JKU JCRNQFKRNQDKQPVKE, CUEQO[EGVQWU URGEKGU KU FKOQTRJKE CPF FKHHGTGPVKCVGU KPVQ [GCUV, RUGWFQO[EGNKWO CPF VTWG UGRVCVG O[EGNKCN HQTOU. +V KU CDNG VQ WUG PQTOCN J[FTQECTDQPU CPF XCTKQWU HCVU CU ECTDQP UQWTEGU, YJKEJ JCU HQTOGF VJG DCUKU QH XCTKQWU KPFWUVTKCN CRRNKECVKQPU. +PKVKCN KPVGTGUV HQEWUGF QP VJG RTQFWEVKQP QH UKPING EGNN RTQVGKPU HTQO PQTOCN RCTCHHKPU, FWTKPI YJKEJ C DCUKE MPQYNGFIG QP JKIJ UECNG, JKIJ FGPUKV[ HGTOGPVCVKQP YCU FGXGNQRGF. 5GEQPF, UKPEG UGXGTCN KUQNCVGU GZETGVG QTICPKE CEKFU KP VJGKT ITQYVJ OGFKWO, QTICPKE CEKF RTQFWEVKQP HTQO XCTKQWU EJGCR ECTDQP UQWTEGU YCU FGXGNQRGF: HQT GZCORNG EKVTKE CEKF RTQFWEVKQP HTQO TCRG UGGF QKN KU UVKNN KORNGOGPVGF QP C NCTIG UECNG VQFC[ D[ VJG #TEJGT &CPKGNU /KFNCPF EQORCP[ (&GECVWT, +NN., 75#ù. &GXGNQROGPV QH VJGUG CRRNKECVKQPU VTKIIGTGF CNUQ DKQUCHGV[ VGUVU QP VJKU [GCUV. Y. lipolytica UVTCKPU WUGF KP VJG CDQXG RTQEGUUGU, YJKEJ CNN TGEGKXGF C )4#5 UVCVWU, Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
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CTG PQP RCVJQIGPKE HQT OCP CPF CPKOCN CU CTG RTGUWOCDN[ OQUV UVTCKPU KUQNCVGF HTQO PCVWTG, RTGUWOCDN[ DGECWUG VJG[ CTG WPCDNG VQ ITQY CV VGORGTCVWTGU CDQXG 3ú¸-3÷¸%. 4GEGPV CRRNKECVKQPU EQPEGTP DKQEQPXGTUKQP QH HCVV[ CEKFU (2CIQV GV CN. ý99ø; öCEJG GV CN. ú00ýù CPF OCPWHCEVWTKPI QH JGVGTQNQIQWU RTQVGKPU, C TGEGPVN[ TGXKGYGF VQRKE (/CF\CM GV CN. ú003ù. #ECFGOKE KPVGTGUV KP VJKU [GCUV UVCTVGF KP VJG GCTN[ ý9õ0•U, HKTUV YKVJ VJG FGXGNQROGPV QH GHHKEKGPV IGPGVKE VQQNU CPF KPDTGF UVTCKPU. 8CTKQWU OGVCDQNKE RTQEGUUGU YGTG VJGP KPXGUVKICVGF (J[FTQECTDQP WVKNKUCVKQP, QTICPKE CEKF, CPF COKPQ CEKF OGVCDQNKUOù, DQVJ DKQEJGOKECNN[ CPF IGPGVKECNN[ (*GUNQV ý990ù. 5KPEG VJG ý9ø0•U, KPVGTGUV GZVGPFGF VQ VJG FGXGNQROGPV QH IGPGVKE GPIKPGGTKPI VQQNU CPF VJGP VQ UVWFKGU QH EGNNWNCT RTQEGUUGU NKMG RTQVGKP UGETGVKQP CPF RGTQZKUQOG DKQIGPGUKU, OQTRJQIGPGUKU CPF UKIPCN VTCPUFWEVKQP RCVJYC[U ($CTVJ CPF )CKNNCTFKP ý99õù. /QTG TGEGPV FGXGNQROGPVU, NKMG OKVQEJQPFTKCN EQORNGZ + DKQIGPGUKU QT IGPQOG UVTWEVWTG CTG TCRKFN[ GZRCPFKPI CPF YKNN DG TGXKGYGF DGNQY. +P CNN VJGUG HKGNFU, Y. lipolytica RTQXGF VQ DG CP QTKIKPCN CPF GZEKVKPI OQFGN: COQPI IGPGVKECNN[ COGPCDNG PQP-EQPXGPVKQPCN [GCUVU, KV KU VCZQPQOKECNN[ VJG OQUV FKUVCPV QPG HTQO S. cerevisiae (5QWEKGV GV CN. ú000ù, CPF VJWU RTQXKFGU WU YKVJ CP WPTKXCNNGF RQKPV QH XKGY HQT CUUGUUKPI GXQNWVKQP CPF EQPUGTXCVKQP QH DCUKE DKQNQIKECN RTQEGUUGU COQPI JGOKCUEQO[EGVQWU [GCUVU.
8.2 )GPGVKEU CPF IGPGVKE VQQNU ©CTN[ FGXGNQROGPVU QH IGPGVKEU CPF IGPGVKE VQQNU KP Yarrowia lipolytica (HQTOGTN[ Candida, Endomycopsis QT Saccharomycopsis lipolyticaù, CU YGNN CU VJG FGUETKRVKQP QH UVTCKPU, NKDTCTKGU CPF RTQVQEQNU HQT IGPGVKE CPF TGEQODKPCPV &û# VGEJPKSWGU JCXG DGGP EQXGTGF KP FGVCKN KP C RTGXKQWU TGXKGY ($CTVJ CPF )CKNNCTFKP ý99üù. 6JKU UGEVKQP YKNN GORJCUKUG TGEGPV FGXGNQROGPVU. 8.2.1.)GPGVKEU 8.2.1.1 Breeding programs Y. lipolytica KU C JGVGTQVJCNNKE [GCUV VJCV GZKUVU KP PCVWTG WPFGT VYQ OCVKPI V[RGU, FGUKIPCVGF CU /CV# CPF /CV$ (UGG ý.ý.úù. 5VCDNG FKRNQKFU CTG QEECUKQPCNN[ HQWPF KP VJG YKNF QT ECP DG ETGCVGF KP XKVTQ D[ HQTEGF OCVKPI DGVYGGP EQORCVKDNG JCRNQV[RGU. $TGGFKPI RTQITCOU CPF KORTQXGF RTQEGFWTGU HQT OCVKPI CPF URQTWNCVKQP ITGCVN[ HCEKNKVCVGF OCPKRWNCVKQP QH NCDQTCVQT[ UVTCKPU ($CTVJ CPF )CKNNCTFKP ý99üù. 6JG NCUV RCRGT QP VJG IGPGVKE OCR YCU RWDNKUJGF KP ý9øú (1IT[F\KCM GV CN. ý9øúù, CPF NKVVNG JCU DGGP CFFGF QT GZVGPFGF UKPEG VJGP. )GPG CUUKIPOGPV D[ J[DTKFK\CVKQP VQ RWNUG HKGNF IGN UGRCTCVGF EJTQOQUQOGU YCU TGRQTVGF KPKVKCNN[ KP ý99õ (%CUCTGIQNC GV CN. ý99õù: QPN[ CTQWPF ø0 IGPGU JCXG RTGUGPVN[ DGGP CUUKIPGF VQ QPG QH VJG UKZ EJTQOQUQOGU QH VJKU URGEKGU (UGG ý.3ù. &KHHGTGPV KPDTGF NKPGU CTG CXCKNCDNG, YJKEJ CTG RQQTN[ KPVGTHGTVKNG ($CTVJ CPF )CKNNCTFKP ý99üù, DWV UVCPFCTF
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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IGPGVKE VGUVU ECP DG GCUKN[ EQPFWEVGF YKVJKP GCEJ NKPG, KPENWFKPI EQORNGOGPVCVKQP CPF NKPMCIG UVWFKGU D[ TCPFQO URQTG CPCN[UKU QT VGVTCF FKUUGEVKQP. 8.2.1.2 Mating type genes and mating pathway /CVKPI V[RG NQEK JCXG DGGP ENQPGF: GCEJ # CPF $ EGNNU ECTT[ C UKPING NQEWU QP EJTQOQUQOG +++, PGZV VQ VJG SLA2 IGPG (þWTKUEJMQ GV CN.ý999; 4. 4QUCU óWKLCPQ, WPRWDNKUJGFù. 7PNKMG KP S. cerevisiae CPF Kluyveromyces lactis, VJGTG CTG PQ UKNGPV OCVKPI-V[RG ECUUGVVGU KP VJG IGPQOG, C UKVWCVKQP CNUQ HQWPF KP C. albicans (*WNN CPF ÿQJPUQP ý999ù. ©CEJ NQEWU ECTTKGU VYQ 14ôU: KP VJG ECUG QH MATA, MATA1 KU TGSWKTGF HQT OCVKPI, YJGTGCU MATA2 TGRTGUUGU URQTWNCVKQP (þWTKUEJMQ GV CN. ý999ù. /CV#ýR UJCTGU PQ UKOKNCTKV[ YKVJ S. cerevisiae /CVCýR, YJGTGCU /CV#úR KU QPN[ FKUVCPVN[ TGNCVGF VQ 5E/CVCúR, DQVJ CTG ENQUGT VQ HWPICN OCVKPI RTQVGKPU (WPRWDNKUJGFù. /CV$ýR CRRGCTU TGNCVGF VQ Sc/CVCNRJCúR, YJGTGCU /CV$úR KU FKUVCPVN[ TGNCVGF VQ Sc/CVCNRJCýR (WPRWDNKUJGFù. #V NGCUV QPG EQR[ QH C RTG-RTQRJGTQOQPG GPEQFKPI IGPG TGNCVGF VQ S. cerevisiae /ôCNRJCR GZKUVU KP VJG IGPQOG, CPF VJG CUUQEKCVGF /#2þ UKIPCNNKPI RCVJYC[ GZKUVU CPF JCU DGGP RCTVKCNN[ UVWFKGF (UGG 3.òù. 8.2.2 )GPGVKE GPIKPGGTKPI 8.2.2.1 Transformation and vectors +PVGITCVKXG VTCPUHQTOCVKQP KU CEJKGXGF D[ VJG NKVJKWO CEGVCVG OGVJQF WUKPI NKPGCTKUGF RNCUOKFU, YJKEJ WPFGTIQ JQOQNQIQWU TGEQODKPCVKQP KPVQ VJG EJTQOQUQOG. 6TCPUHQTOCVKQP HTGSWGPEKGU QH WR VQ ý0ü RGT OKETQITCO QH &û# YGTG TGRQTVGF OCMKPI NKDTCT[ UETGGPKPI RQUUKDNG. ©NGEVTQRQTCVKQP RTQVQEQNU CTG RTGHGTTGF HQT TGRNKECVKPI RNCUOKFU CPF C QPG UVGR OGVJQF HTQO UKPING EQNQPKGU JCU DGGP FGXGNQRGF (%JGP GV CN. ý99õCù. 5GCTEJGU HQT TGUKFGPV RNCUOKFU CPF HQT ARS (#WVQPQOQWUN[ 4GRNKECVKPI 5GSWGPEGù XGEVQTU YGTG KPKVKCNN[ WPUWEEGUUHWN. +V VWTPGF QWV VJCV ARSs CNQPG CTG WPCDNG VQ UWRRQTV GZVTCEJTQOQUQOCN TGRNKECVKQP WPNGUU VJG[ CTG CUUQEKCVGF YKVJ C CEN UGSWGPEG (ôQWTPKGT GV CN. ý993; 8GTPKU GV CN. ý99õù. ARSs CTG UJQTV UGSWGPEGU YKVJQWV PQVKEGCDNG EQPUGTXGF HGCVWTGU, YJKEJ RTQOQVG KPKVKCVKQP QH &û# TGRNKECVKQP CV EJTQOQUQOCN UKVGU (8GTPKU GV CN. ý999ù. ôWPEVKQPCN CEN UGSWGPEGU CTG UKOKNCT KP UK\G (DWV PQV UGSWGPEGù VQ VJQUG QH S. cerevisiae. ôKXG QH VJG ü EGPVTQOGTG TGIKQPU JCXG DGGP KUQNCVGF CPF UGSWGPEGF; KPVGTGUVKPIN[, VJG[ FQ PQV UJQY CP[ EQPUGTXGF UGSWGPEG OQVKH. 2NCUOKFU ECTT[KPI CEN/ARS TGRNKECVG GZVTCEJTQOQUCOCN[ YKVJ C NQY EQR[ PWODGT CPF CTG SWKVG UVCDNG OKVQVKECNN[, VJG[ GZJKDKV OGPFGNKCP UGITGICVKQP CV OGKQUKU, NKMG S. cerevisiae CEN RNCUOKFU (8GTPKU GV CN. ú00ýù. 6JGUG RNCUOKFU JCXG DGGP WUGF KP RNCUOKF UJWHHNGU VQ KUQNCVG EQPFKVKQPCN OWVCVKQPU CPF GZVTCIGPKE UWRRTGUUQTU. /WNVKEQR[ KPVGITCVKXG XGEVQTU JCXG DGGP FGXGNQRGF. 6JG QTKIKPCN XGEVQTU ECTTKGF C FGHGEVKXG URA3 OCTMGT, CP KPUGTV QH T&û#, CPF ENQPKPI UKVGU HQT VJG IGPG QH KPVGTGUV (ñG &CNN GV CN. ý99òù. öKVJ UWEJ XGEVQTU, EQR[ PWODGTU CU JKIJ CU ü0 JCXG
ú30
). $CTVJ GV CN.
DGGP TGRQTVGF. #P CNVGTPCVKXG OWNVKEQR[ KPVGITCVKXG U[UVGO YCU FGXGNQRGF DCUGF QP VJG FGHGEVKXG URA3 IGPG CPF VJG ü0ù KP VJG IGPQOG, YJGTGCU KP
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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VJG URA3 IGPG FGNGVGF. 6JG UJQTV OWVCIGPGUKU ECUUGVVG KPUGTVU NCTIGN[ CV TCPFQO CPF VJG VCIIGF IGPGU CTG TGCFKN[ KFGPVKHKGF (/CWGTUDGTIGT GV CN. ú00ýù. 8.2.3 )GPQOKEU 6JG IGPQOG QH Y. lipolytica EQPUKUVU KP UKZ EJTQOQUQOGU, VQVCNNKPI EC. ú0 /D QH &û#, YJKEJ CTG JKIJN[ RQN[OQTRJKE COQPI FKHHGTGPV KUQNCVGU (%CUCTGIQNC GV CN. ý99õù. 4GRGCVGF UGSWGPEGU EQPUKUV KP QPG VQ UKZ UWDVGNQOGTKE ENWUVGTU QH T&û# (ôQWTPKGT GV CN. ý9øü; XCP *GGTKMJWK\GP GV CN. ý9ø÷ù, CV NGCUV VYQ TGVTQVTCPURQUQPU QH VJG 6[3 HCOKN[ (ûGWXGINKUG GV CN. ú00ú; 5EJOKF-$GTIGT GV CN. ý99òù CPF C ñKPG GNGOGPV, ðKNK (%CUCTGIQNC GV CN. ú00úù. ñKPG GNGOGPVU CTG TGVTQVTCPURQUQPU YKVJQWV ñ64 YJKEJ CTG CDUGPV HTQO S. cerevisiae CPF S. pombe IGPQOGU, DWV CTG CDWPFCPV KP JKIJGT GWMCT[QVGU CPF FQ QEEWT KP HKNCOGPVQWU HWPIK. 4GEGPVN[, QPG ñKPG YCU FGUETKDGF KP C. albicans ()QQFYKP GV CN. ú00ýù, UWIIGUVKPI VJCV ñKPGU YGTG RTGUGPV KP VJG CPEGUVQT QH OQFGTP [GCUVU CPF OC[ JCXG RNC[GF CP KORQTVCPV TQNG KP IGPQOG UJCRKPI. #TQWPF ý÷0 PQP-TGFWPFCPV GPVTKGU EWTTGPVN[ GZKUV KP )GP$CPM, JCNH QH YJKEJ DGKPI CUUKIPGF VQ C EJTQOQUQOG, CPF VJG EQORNGVG OKVQEJQPFTKCN &û# UGSWGPEG KU CXCKNCDNG (þGTUEJGT GV CN. ú00ýCù; UGG DGNQYù. #DQWV ý3ï QH VJG IGPGU EQPVCKP QPG QT QEECUKQPCNN[ VYQ KPVTQPU, QHVGP QH C TGNCVKXGN[ NCTIG UK\G (ò00PV, UGG CNUQ ($QP GV CN. ú003ùù. #P GZRNQTCVQT[ UGSWGPEKPI RTQITCO ECNNGF )oPQNGXWTGU (5QWEKGV GV CN ú000ù; UGG CNUQ %JCRVGT úù JGNRGF KFGPVKH[KPI ýúú9 IGPGU COQPI ò9ò0 ý MD TCPFQO UGSWGPEG VCIU VQVCNNKPI ò,9 /D (%CUCTGIQNC GV CN. ú000ù. +V EQPHKTOGF VJCV Y. lipolytica KU XGT[ FKUVCPV HTQO S. cerevisiae CPF CEVWCNN[ HTQO OQUV QVJGT JGOKCUEQO[EGVQWU [GCUVU ($CTPU GV CN. ý99ýù. # RCTVKCNN[ QTFGTGF $#% NKDTCT[ KU CXCKNCDNG CPF VJG HWNN IGPQOKE UGSWGPEG KU EWTTGPVN[ DGKPI CUUGODNGF D[ VJG ôTGPEJ ûCVKQPCN 5GSWGPEKPI %GPVTG ()GPQUEQRG, ©XT[, ôTCPEGù.
8.3 Yarrowia lipolytica CU C OQFGN HQT RTQVGKP UGETGVKQP Yarrowia lipolytica UGETGVGU UGXGTCN RTQVGKPU KPVQ KVU ITQYVJ OGFKWO CEEQTFKPI VQ GPXKTQPOGPVCN EQPFKVKQPU, KPENWFKPI CV NGCUV VYQ RTQVGCUGU (CP CNMCNKPG GZVTCEGNNWNCT RTQVGCUG =#©2? CPF CP CEKF GZVTCEGNNWNCT RTQVGCUG =#î2?ù, UGXGTCN NKRCUGU CPF RJQURJCVCUGU, C 4ûCUG CPF CP GUVGTCUG ($CTVJ CPF )CKNNCTFKP ý99õù. 6JKU RTQORVGF KVU WVKNK\CVKQP CU CP GZRTGUUKQP U[UVGO HQT VJG UGETGVKQP QH HQTGKIP RTQVGKPU, C UWEEGUUHWN FQOCKP VJCV JCU DGGP TGEGPVN[ TGXKGYGF (&QOKPIWG\ GV CN. ý99ø; /CF\CM GV CN. ú003ù: OQTG VJCP ò0 JGVGTQNQIQWU RTQVGKPU JCXG DGGP GZRTGUUGF UWEEGUUHWNN[ KP VJKU JQUV. +P UGXGTCN ECUGU, JQYGXGT, GZRTGUUKQP NGXGNU YGTG FKUCRRQKPVKPIN[ NQY, YJKEJ RTQORVGF UGXGTCN ITQWRU VQ FQ CP KP FGRVJ CPCN[UKU QH VJG UGETGVKQP RCVJYC[ CPF QH RQVGPVKCN UGETGVQT[ DQVVNGPGEMU. 6JG GZKUVGPEG QH C JKIJN[ GZRTGUUGF UGETGVQT[ RTQVGKP (#©2ù, YJKEJ WPFGTIQGU C YGNN-FGHKPGF RTQEGUUKPI CNQPI KVU UGETGVQT[ TQWVG, JCU ITGCVN[ HCEKNKVCVGF VJG CPCN[UKU QH UGETGVQT[ OWVCPVU (*G GV CN. ý99ú; ðCXGT GV CN. ý99úù. 6JG HKTUV KPVGTOGFKCVG FGVGEVGF KU C IN[EQU[NCVGF, UKIPCN UGSWGPEG-RTQEGUUGF RTGEWTUQT ECTT[KPI C NCTIG RTQTGIKQP,
ú3ú
). $CTVJ GV CN.
YJKEJ KU RTQEGUUGF KP VJG )QNIK CRRCTCVWU CPF VJGP UGETGVGF KP VJG ITQYVJ OGFKWO YKVJ C VTCPUKV VKOG QH CDQWV ú OKP. /QUV UVWFKGU QH VJG UGETGVQT[ RCVJYC[ YGTG DCUGF QP IGPGVKE UETGGPU EGPVTGF QP VJG GCTN[ UGETGVQT[ UVGRU, NGCFKPI HTQO VJG TKDQUQOGU VQ VJG ©4 NWOGP. 8.3.1 ÿTGFQOKPCPEG QH C EQVTCPUNCVKQPCN VTCPUNQECVKQP RCVJYC[ HTQO E[VQRNCUO VQ ©4 6YQ RCVJYC[U VCTIGVKPI UGETGVQT[ RQN[RGRVKFGU VQ VJG GPFQRNCUOKE TGVKEWNWO (©4ù JCXG DGGP FGUETKDGF KP GWMCT[QVGU: (Kù C RQUV-VTCPUNCVKQPCN RCVJYC[, YJGTG HWNN NGPIVJ E[VQRNCUOKE RTGEWTUQTU, CUUKUVGF D[ EJCRGTQPGU, CTG VCTIGVGF VQ C RQUVVTCPUNCVKQPCN VTCPUNQECVKQP RQTG CPF (KKù C EQVTCPUNCVKQPCN RCVJYC[, YJGTG C 5KIPCN 4GEQIPKVKQP 2CTVKENG (542ù DKPFU VQ VJG TKDQUQOG CPF UNQYU FQYP VTCPUNCVKQP CPF FQEMU VJG YJQNG EQORNGZ VQ CP ©4 DQWPF 542 TGEGRVQT HTQO YJKEJ VJG PCUEGPV RQN[RGVKFG KU VTCPUHGTTGF VQ C VTCPUNQECVKQP RQTG, YJGTG VTCPUNCVKQP TGUWOGU VQIGVJGT YKVJ VTCPUNQECVKQP KPVQ VJG ©4 NWOGP. +P Yarrowia, EQORQPGPVU DGNQPIKPI VQ GCEJ QH VJGUG RCVJYC[U JCXG DGGP KFGPVKHKGF. ôQT VJG RQUV-VTCPUNCVKQPCN RCVJYC[, SEC61, SEC62, SEC632 YGTG ENQPGF UGXGTCN [GCTU CIQ ($TQWIJVQP GV CN. ý99õ; 5YGPPGP GV CN. ý99õù. ôQT VJG EQVTCPUNCVKQPCN RCVJYC[, EQORQPGPVU QH VJG 542 YGTG EJCTCEVGTKUGF: SCR1, SCR2, SRP54, SEC65, CPF SRP101 GPEQFKPI VJG 542 TGEGRVQT (*G GV CN. ý99ú; þKO CPF 1IT[F\KCM ú000; ñGG CPF 1IT[F\KCM ý99õ; 5CPEJG\ GV CN. ý99õ; ðCXGT GV CN. ý99úù. +PVGTGUVKPIN[, VJG õ5 4û# EQORQPGPV QH VJG 542 QH ðCTTQYKC, YJKEJ KU WPWUWCN KP S. cerevisiae, YCU HQWPF VQ ENQUGN[ TGUGODNG KVU JWOCP CPF S. pombe JQOQNQIWGU CPF FGNGVKQPU CHHGEVKPI SCR QT SRP IGPGU YGTG WUWCNN[ NGVJCN EQPVTCT[ VQ VJG UKVWCVKQP QDUGTXGF KP S. cerevisiae. #NVJQWIJ DQVJ RCVJYC[U CRRGCT VQ DG HWPEVKQPCN KP Yarrowia, VJG EQVTCPUNCVKQPCN TQWVG UGGOU VQ DG RTGFQOKPCPV, EQPVTCT[ VQ S. cerevisiae. 2WNUG-EJCUG CPCN[UKU QH #©2 VTCPUNQECVKQP KPKVKCNN[ UWIIGUVGF VJCV #©2 VTCPUNQECVKQP YCU GUUGPVKCNN[ EQVTCPUNCVKQPCN (/CVQDC GV CN. ý9øøù, C HKPFKPI NCVGT GZVGPFGF VQ OQUV UGETGVQT[ RTQVGKPU: OQTG VJCP õ÷ï QH VTCPUNQECVKQP RQTGU CTG TKDQUQOG DQWPF CPF VJWU QH VJG RQUV-VTCPUNCVKQPCN V[RG ($QKUTCOG GV CN. ý99øù. ôWTVJGTOQTG, EQPFKVKQPCN OWVCVKQPU QH VJG 542 EQORQPGPVU NKMG Ucr2-II-13 UVTQPIN[ FGETGCUG #©2 VTCPUNCVKQP GHHKEKGPE[ YKVJQWV CEEWOWNCVKQP QH E[VQRNCUOKE RTGEWTUQTU, KPFKECVKPI VKIJV EQWRNKPI QH VTCPUNCVKQP CPF VTCPUNQECVKQP. 6JKU UKVWCVKQP KU TCVJGT UKOKNCT VQ VJG QPG FGUETKDGF HQT JKIJGT GWMCT[QVGU. +V RTQORVGF C JWPV HQT QVJGT EQORQPGPVU QH VJKU RCVJYC[. 8.3.2 5NU1R, RTQVQV[RG QH C PGY #&ÿ/#6ÿ GZEJCPIG HCEVQT HCOKN[ HQT GWMCT[QVKE *UR70R 5[PVJGVKE NGVJCN OWVCPVU CPF UWRRTGUUQTU QH VJG EQPFKVKQPCN OWVCVKQP scr2-II-13, YJKEJ KORCKTU VJG HWPEVKQP QH VJG 542 YGTG HKTUV KUQNCVGF CPF CPCN[UGF.
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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SLS1 YCU KFGPVKHKGF KP C U[PVJGVKE NGVJCN UETGGP YKVJ scr2-II-13 ($QKUTCOG GV CN. ý99üù CU C PGY IGPG YKVJQWV RTGFKEVCDNG HWPEVKQP, DWV DGNQPIKPI VQ C HCOKN[ QH IGPGU EQPUGTXGF HTQO [GCUV VQ OCP. +P C YKNF V[RG DCEMITQWPF, VJG UWRRTGUUQT OWVCVKQP sls1-1 CU YGNN CU VJG EQORNGVG FGNGVKQP •sls1 YGTG UJQYP VQ EQPHGT C FGHGEV KP EQVTCPUNCVKQPCN VTCPUNQECVKQP. +V YCU NCVGT UJQYP VJCV sls1-1 NGCFU VQ 5NUýR FGUVCDKNK\CVKQP CPF TCRKF RTQVGQN[UKU CV VJG PQP-RGTOKUUKXG VGORGTCVWTG. #U UJQYP D[ KOOWPQNQECNKUCVKQP CPF UWDEGNNWNCT HTCEVKQPCVKQP, 5NUýR KU C NWOGPCN EQORQPGPV QH VJG ©4, YKVJ CP û-VGTOKPCN UKIPCN UGSWGPEG CPF C %-VGTOKPCN ©4 TGVGPVKQP VGVTCRGRVKFG. ôTCEVKQPCVKQP UVWFKGU UJQYGF VJCV 5NUýR YCU CUUQEKCVGF YKVJ VJG 5GEüýR EQVTCPUNCVKQPCN VTCPUNQEQP. 6JKU CUUQEKCVKQP KU FKUTWRVGF KP VJG sls1-5 OWVCPV ($QKUTCOG GV CN. ý99øù KPFWEGF D[ UKVG-FKTGEVGF OWVCIGPGUKU KP C TGIKQP EQPUGTXGF CETQUU VJG 5NUýR HCOKN[. $[ VYQ-J[DTKF UETGGPKPI, CPF EQPHKTOGF D[ EQKOOWPQRTGEKRKVCVKQP, KV YCU FGOQPUVTCVGF VJCV 5NUýR DKPFU VQ VJG #62CUG FQOCKP QH þCTúR, VJG OCLQT JURõ0 QH VJG ©4 (þCDCPK GV CN. ú000ù. 6JGTGHQTG, 5NUýR CRRGCTGF VQ DG C PQXGN TGIWNCVQT QH þCTúR HWPEVKQP (ôKI. ýù.
(KI. 1. 5EJGOCVKE OQFGN HQT 5NUýR KPVGTCEVKQPU YKVJ þCTúR CPF 5GEü3R. þCTúR E[ENGU DGVYGGP CP "QRGP" #62-FGRGPFGPV EQPHQTOCVKQP, YJKEJ CNNQYU NQQUG KPVGTCEVKQPU YKVJ WPHQNFGF RQN[RGRVKFGU CPF C "ENQUGF" #&2-FGRGPFGPV EQPHQTOCVKQP, YJKEJ KPUWTGU C VKIJV DKPFKPI VQ VJG NKPMGF RQN[RGRVKFG. 6JG UYKVEJ HTQO VJG QRGP VQ VJG ENQUGF EQPHQTOCVKQP KU UVKOWNCVGF D[ VJG ÿ FQOCKP QH 5GEü3R, YJKEJ KPETGCUGU VJG #62CUG CEVKXKV[ QH þCTúR. 5NUýR KPFWEGF GZEJCPIG DGVYGGP #&2 DQWPF VQ þCTúR CPF GZVGTPCN #62 KORTQXKPI VJG TGE[ENKPI QH þCTúR CPF VJG TGNGCUG QH VJG DQWPF RQN[RGRVKFG (HTQO /GJFK þCDCPK, 2J& VJGUKUù.
+P VJG E. coli JURõ0í&PCþ U[UVGO, &PCÿ CEVU CU CP GHHGEVQT QH VJG #62CUG &PCþ CEVKXKV[ CPF )TR© CU CP #&2í#62 GZEJCPIG HCEVQT, DQVJ HCEVQTU EQQRGTCVKPI VQ KPETGCUG E[ENKPI QH &PCþ DGVYGGP KVU QRGP #62-DQWPF EQPHQTOCVKQP CPF KVU #&2-DQWPF EQPHQTOCVKQP ENQUGF QP C OKUHQNFGF RQN[RGRVKFG (ôQT TGXKGY, UGG HQT KPUVCPEG (©NNKU CPF *CTVN ý999ùù. +P VJG GWMCT[QVKE ©4 EQORCTVOGPV, JQOQ-
ú3ò
). $CTVJ GV CN.
NQIWGU QH &PCÿ CTG YGNN FGUETKDGF, CNN QH VJGO RQUUGUUKPI C EQOOQP EQPUGTXGF ÿ FQOCKP. ôQT KPUVCPEG, KP Saccharomyces, VJTGG RCTVPGTU YGTG FGUETKDGF: 5GEü3R, ÿGOýR, CPF 5ELý. 1P VJG EQPVTCT[, PQ JQOQNQIWG QH VJG DCEVGTKCN )TR© RTQVGKP HCOKN[ JCF DGGP KFGPVKHKGF KP VJG GWMCT[QVKE YQTNF. $[ in vitro OGVJQFU, YG JCXG UJQYP VJCV VJG S. cerevisiae JQOQNQIWG QH ðN5NUýR CEVU CU CP #&2í#62 GZEJCPIG HCEVQT HQT þCTúR (þCDCPK GV CN. ú000ù, NKMG )TR© HQT &PCþ CNVJQWIJ KV JCU PQ UGSWGPEG UKOKNCTKV[ YKVJ )TR©. 6JGUG TGUWNVU YGTG GZVGPFGF VQ C FKUVCPV E[VQRNCUOKE JQOQNQIWG QH 5NUýR, ôGUýR, YJKEJ GZGTVU CP 5NUýR-NKMG HWPEVKQP, DWV QP E[VQRNCUOKE *URõ0R (þCDCPK GV CN. ú00úù. *QYGXGT, KV TGOCKPU VQ DG GUVCDNKUJGF KP RTGEKUGN[ YJKEJ HWPEVKQP QH þCTúR 5NUýR KU KPXQNXGF. +P Yarrowia, 5NUýR UGGOU VQ DG URGEKHKECNN[ KPXQNXGF KP EQVTCPUNCVKQPCN VTCPUNQECVKQP, CPF YG JCXG PQ GXKFGPEG HQT CP KPXQNXGOGPV KP VJG RQUVVTCPUNCVKQPCN RCVJYC[. 6JG QDUGTXCVKQP VJCV SLS1 QXGTGZRTGUUKQP NGPIVJGPU VJG VTCPUKV QH VJG #©2 RTGEWTUQT VJTQWIJ VJG ©4 EQWNF KPFKECVG VJCV 5NUýR OC[ CNUQ RCTVKEKRCVG KP VJG EJCRGTQPG TQNG QH þCTúR KPUKFG VJG ©4 NWOGP CHVGT VTCPUNQECVKQP. 8.3.3 þVJGT PGY EQORQPGPVU CHHGEVKPI UGETGVKQP Sls2-1 YCU CNUQ KUQNCVGF KP C U[PVJGVKE NGVJCN OWVCPV UETGGP YKVJ scr2-II-13 ($QKUTCOG GV CN. ý999ù. SLS2 GPEQFGU C 9ýú COKPQ CEKF NQPI RQN[RGRVKFG, YJKEJ FKURNC[U C %-VGTOKPCN HCTPGU[NCVKQP UKIPCN. 6JG OWVCVKQP CNQPG JCF CNOQUV PQ GHHGEV QP #©2 VTCPUNQECVKQP CPF KV YCU UJQYP VJCV KVU S. cerevisiae JQOQNQIWG RCY1 YCU KPXQNXGF KP VJG GPFQE[VKE RCVJYC[ (öKGFGTMGJT GV CN. ú000ù, XGT[ HCT HTQO VJG GCTN[ UVGRU QH VJG RCVJYC[. TSR1 CPF TSR3 YGTG KUQNCVGF CU GZVTCIGPKE, OWNVKEQR[-UWRRTGUUQTU QH sls2-II13. $QVJ CTG GUUGPVKCN IGPGU KP Yarrowia. 7RQP C UJKHV CV VJG PQP-RGTOKUUKXG VGORGTCVWTG, DQVJ OWVCVKQPU XGT[ SWKEMN[ KORCKTGF #©2 VTCPUNQECVKQP, KPFKECVKPI VJCV VJG[ OC[ CNVGT C EQORQPGPV NKPMGF VQ VJKU RTQEGUU. TSR1 GPEQFGU C òüý COKPQ CEKF RQN[RGRVKFG YKVJ C VTCPUOGODTCPG FQOCKP (/COQWP GV CN. ý99üù. +V YCU UJQYP VQ KPVGTCEV D[ EQKOOWPQRTGEKRKVCVKQP YKVJ 542 CPF TKDQUQOG EQORQPGPVU. 1WT EWTTGPV OQFGN KU VJCV 6UTýR TGIWNCVGU KPVGTCEVKQP QH VJG 542 CPF VJG TKDQUQOG CV VJG ©4 OGODTCPG (/COQWP GV CN. ý99õù, DWV FGVCKNU CTG UVKNN OKUUKPI. +VU ENQUGUV JQOQNQIWG KP S. cerevisiae YHC8, CNUQ MPQYP CU WSC4, YCU UJQYP VQ DG NQECVGF KP VJG ©4 EQORCTVOGPV D[ KOOWPQHNWQTGUEGPEG (/COQWP GV CN. ý999ù. 6JKU IGPG KU PQV GUUGPVKCN KP 5. EGTGXKUKCG EQPVTCT[ VQ KVU JQOQNQIWG KP Yarrowia, CPF KVU FGNGVKQP CNQPG FQGU PQV EQPHGT CP[ QDXKQWU RJGPQV[RG. +P EQODKPCVKQP YKVJ EQPFKVKQPCN OWVCVKQPU FGHGEVKXG KP VTCPUNQECVKQP (G.I. KP SEC61ù, •yhc8 EQPHGTU C U[PVJGVKE FGHGEVKXG VTCPUNQECVKQP RJGPQV[RG. 6UTýR CPF ðJEøR DGNQPI VQ C RTQVGKP HCOKN[ YJQUG QVJGT OGODGTU CTG TGRQTVGF CU EGNN YCNN UGPUQTU (8GTPC GV CN. ý99õù. 6JGTGHQTG, VJG TQNG QH VJGUG EQORQPGPVU KU UVKNN GPKIOCVKE. TSR3 GPEQFGU C RQN[RGRVKFG YKVJQWV CP[ QDXKQWU OQVKH, DWV KV DGNQPIU VQ C EQPUGTXGF IGPG HCOKN[ HTQO [GCUV VQ OCOOCNU. +V CRRGCTU VQ DG TGNCVGF VQ VJG CEVKP PGVYQTM CU GXKFGPEGF VJTQWIJ VYQ-J[DTKF NKDTCT[ UETGGPKPI CU YGNN CU KO-
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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OWPQHNWQTGUEGPEG UVWFKGU (2. ÿQ[GV, WPRWDNKUJGF FCVCù, CPF OC[ TGRTGUGPV C PGY NKPM DGVYGGP VJG UGETGVQT[ PGVYQTM CPF VJG E[VQUMGNGVVQP. +P UWOOCT[, UETGGPKPI RTQEGFWTGU FGFKECVGF VQ HKPF RCTVPGTU QH 542 CNNQYGF TCVJGT UWTRTKUKPIN[ VQ KFGPVKH[ OWVCVKQPU CHHGEVKPI XGT[ FKHHGTGPV UVGRU QH VJG UGETGVKQP RCVJYC[. 6JKU TGHNGEVU OQUV RTQDCDN[ KPVGITCVKQP QH CNN UGETGVKQP UVGRU CNQPI VJG RCVJYC[: CP[ DNQEM KP VJG RTQEGUU KU VJTQYP DCEM VQ VJG KPKVKCN UVGR, VJG U[PVJGUKU QH VJG UGETGVGF RQN[RGRVKFG. 6JKU KPVGITCVKQP, YJKEJ KU RTQDCDN[ GUUGPVKCN HQT EGNN UWTXKXCN, YCU TGEGPVN[ CRRTQCEJGF KP Saccharomyces cerevisiae D[ C VTCPUETKRVQOG CPCN[UKU QH EGNNU WPFGTIQKPI CP WPHQNFGF RTQVGKP UVTGUU (6TCXGTU GV CN. ú000ù: VJKU UVWF[ JCU KFGPVKHKGF SLS1 CU C VCTIGV HQT VJG 7PHQNFGF 2TQVGKP 4GURQPUG (724ù TGIWNCVKQP. 8.3.ý (WPEVKQP QH VJG S. cerevisiae JQOQNQIWGU QH SEC IGPGU # HGY CVVGORVU YGTG EQPFWEVGF CV KPXGUVKICVKPI VJG HWPEVKQP QH JQOQNQIWGU QH S. cerevisiae IGPGU KPXQNXGF KP VTCPUNQECVKQP ($QKUTCOG GV CN. ú00úù. 5GEüý¾ KP OCOOCNU, 5DJýRí5DJúR KP Saccharomyces KU C UOCNN EQORQPGPV QH VJG VTCPUNQECVKQP RQTG. +VU TQNG KU WPMPQYP WPVKN PQY. 6JG Yarrowia SBH1 IGPG YCU ENQPGF D[ EQORNGOGPVCVKQP QH VJG VJGTOQUGPUKVKXKV[ QH CP •sbh1,•sbh2 UVTCKP QH S. cerevisiae WUKPI C Y. lipolytica E&û# NKDTCT[. YlSBH1 FGNGVKQP EQPHGTTGF PQ RJGPQV[RG. 6JGTGHQTG, RCTVPGTU YGTG UQWIJV D[ C VYQ-J[DTKF NKDTCT[ UETGGPKPI. # UKPING RCTVPGT YCU KFGPVKHKGF, ECNPGZKP (%PZRýù. %CNPGZKP KU C EQORQPGPV QH VJG ©4 OGODTCPG CPF KV KU VJQWIJV VQ RNC[ VJG TQNG QH C NGEVKP, DKPFKPI OKUHQNFGF IN[EQU[NCVGF RQN[RGRVKFGU. 6JG QDUGTXGF KPVGTCEVKQP QH 5DJýR YKVJ %PZýR DTKPIU ECNPGZKP PGCT VJG VTCPUNQECVKQP RQTG, CPF TCKUGU C PGY SWGUVKQP: FQGU ECNPGZKP DKPF PCUEGPV RQN[RGRVKFGU DGHQTG QT CHVGT VJGKT IN[EQU[NCVKQP! ôWTVJGT UVWF[ QH VJKU RCTVPGTUJKR KP Yarrowia EQWNF OQFKH[ QWT XKGY QH VJG HWPEVKQP QH ECNPGZKP. 1XGTNQQMKPI VJG GZCORNGU FGUETKDGF CDQXG, Yarrowia RTQXGU VQ DG C XGT[ HTWKVHWN QTICPKUO HQT VJG UVWF[ QH VJG UGETGVKQP RCVJYC[. +V KU VJG QPN[ WPKEGNNWNCT OKETQQTICPKUO YJGTG VTCPUNQECVKQP YCU UJQYP VQ DG OCKPN[ EQVTCPUNCVKQPCN CU KP OCOOCNKCP EGNNU. Yarrowia CRRGCTU VJWU CU C EQORNGOGPV VQ Saccharomyces YJQUG VTCPUNQECVKQP KU OCKPN[ RQUV-VTCPUNCVKQPCN. &KHHGTGPEGU KP VJG GCTN[ UVGRU QH VJG UGETGVKQP RCVJYC[ RTQDCDN[ GZRNCKP YJ[ EQORQPGPVU KFGPVKHKGF KP Yarrowia D[ IGPGVKE UETGGPU CTG FKHHGTGPV HTQO VJQUG HQWPF KP Saccharomyces. +V OC[ CNUQ GZRNCKP YJ[ EGTVCKP RQN[RGRVKFGU, PQTOCNN[ UGETGVGF RQUV-VTCPUNCVKQPCNN[ KP VJGKT QTKIKPCN JQUV, CTG GCUKGT VQ OCPWHCEVWTG KP VJKU QTICPKUO, CPF UWIIGUV JKPVU QP JQY VQ OCPKRWNCVG VJKU RCVJYC[ VQ HCEKNKVCVG RTQFWEVKQP QH TGNWEVCPV RQN[RGRVKFGU.
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). $CTVJ GV CN.
8.ý Yarrowia lipolytica CU C OQTRJQIGPGVKE OQFGN 8.ý.1 üQTRJQIGPGUKU KP Yarrowia lipolytica /CP[ HWPIK CTG CDNG VQ UYKVEJ DGVYGGP C WPKEGNNWNCT [GCUV HQTO CPF C HKNCOGPVQWU HQTO KP TGURQPUG VQ GPXKTQPOGPVCN CPF PWVTKVKQPCN UKIPCNU. 4GUGCTEJ KP [GCUV FKOQTRJKUO JCU DGPGHKVGF HTQO VJG HCEV VJCV VJG OQFGN [GCUV Saccharomyces cerevisiae WPFGTIQGU C FKOQTRJKE VTCPUKVKQP VQ HKNCOGPVQWU ITQYVJ (RUGWFQJ[RJCN FKHHGTGPVKCVKQPù KP TGURQPUG VQ PWVTKGPV UKIPCNU. 6YQ JWOCP HWPICN RCVJQIGPU, Candida albicans CPF Cryptococcus neoformans, CPF QPG RNCPV HWPICN RCVJQIGP Ustilago maydis CTG CNUQ UWKVCDNG OQFGNU HQT CPCN[\KPI FKOQTRJKUO (ñGPIGNGT GV CN. ú000ù. Y. lipolytica KU CDNG VQ WPFGTIQ C VTWG [GCUV-J[RJC VTCPUKVKQP KP DQVJ JCRNQKF CPF FKRNQKF EGNNU (*GTTGTQ GV CN. ý999ù. +V FKURNC[U C RQNCT DWFFKPI RCVVGTP KP FKRNQKF CPF JCRNQKF EGNNU QH DQVJ OCVKPI V[RGU, WPNKMG VJG UKVWCVKQP TGRQTVGF KP S. cerevisiae QT C. albicans (*GTTGTQ GV CN. ý999ù. 6JG [GCUV-J[RJC VTCPUKVKQP ECP DG UKORN[ KPFWEGF D[ GKVJGT EJCPIKPI VJG ECTDQP UQWTEG INWEQUG HQT ûCEGV[NINWEQUCOKPG, QT D[ CFFKPI UGTWO VQ VJG EWNVWTG OGFKWO (þKO GV CN. ú000; 2GTG\-%CORQ CPF &QOKPIWG\ ú00ýù. #ICT KPXCUKQP QEEWTU TGCFKN[ QP EQORNGVG UQNKF OGFKC (6TGVQP GV CN. ú000ù. 8.ý.2 ©ZRGTKOGPVCN CRRTQCEJGU HQT VJG OQNGEWNCT ENQPKPI QH IGPGU KPXQNXGF KP VJG [GCUV-J[RJC VTCPUKVKQP KP Yarrowia lipolytica 5KPEG OWVCPVU CHHGEVGF KP OQTRJQIGPGUKU FKURNC[ C ENGCT RJGPQV[RG QP RNVGU, KUQNCVKQP QH OQPQOQTRJKE OWVCPVU CPF VJGKT EQORNGOGPVCVKQP YKVJ IGPQOKE NKDTCTKGU YCU KPKVKCNN[ CVVGORVGF. #FFKVKQPCN OGVJQFU YGTG NCVGT KORNGOGPVGF, NKMG VTCPURQUQP-OGFKCVGF OWVCIGPGUKU, O4û# FKHHGTGPVKCN FKURNC[, FKHHGTGPVKCN J[DTKFK\CVKQP UETGGPKPI, CPF VJG ENQPKPI QH JQOQNQIQWU IGPGU QH S. cerevisiae CPF C. albicans WUKPI FGIGPGTCVG QNKIQPWENGQVKFGU KP EQPUGTXGF IGPG TGIKQPU. 6JCPMU VQ VJG EQORNGVKQP QH VJG S. cerevisiae, C. albicans CPF 5. pombe IGPQOGU, CPF VQ VJG )GPQNGXWTG RTQLGEV, in silico UGCTEJ HQT JQOQNQIWGU KU PQY CNUQ RQUUKDNG. 8.ý.3 +FGPVKHKECVKQP CPF EJCTCEVGTK\CVKQP QH Y. lipolytica OQTRJQIGPGVKE IGPGU +P QTFGT VQ KFGPVKH[ VJG OQNGEWNCT OGEJCPKUOU WPFGTN[KPI VJG FKOQTRJKE VTCPUKVKQP, C XKUWCN UETGGPKPI QP RNCVGU HQT Y. lipolytica ôKN- (HKNCOGPVCVKQP OKPWU OWVCPVUù JCU DGGP FGXGNQRGF (ôQWTPKGT GV CN. ý99ý; 6QTTGU-)W\OCP CPF &QOKPIWG\ ý99õ; 6QTTGU-)W\OCP GV CN. ý99üù. ûQTOCN Y. lipolytica UVTCKPU HQTO TQWIJUWTHCEGF EQNQPKGU QP ð©& (ôKI. ú#ù CPF CHVGT OWVCIGPGUKU, UOQQVJ PQPHKNCOGPVQWU EQNQPKGU ECP DG TGCFKN[ KUQNCVGF (ôKI. ú$ù.
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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(KI. 2. %QNQPKCN OQTRJQNQI[ KP ð. NKRQN[VKEC. (#ù: YKNF V[RG, YTKPMNGF EQNQP[; (($ù ôKN- OWVCPV, UOQQVJ, OQTRJQNQI[.
/WVCPVU DGNQPIKPI VQ CV NGCUV UGXGP EQORNGOGPVCVKQP ITQWRU YGTG KUQNCVGF CHVGT ©/5 OWVCIGPGUKU. 6Q FCVG, QPN[ QPG QH VJGO, HOY1, JCU DGGP EQORNGOGPVGF (6QTTGU-)W\OCP CPF &QOKPIWG\ ý99õ; 6QTTGU-)W\OCP GV CN. ý99üù &QOKPIWG\, WPRWDNKUJGF YQTMù. 6JTQWIJ UKOKNCT UETGGPU, CPQVJGT HKXG IGPGU - MHY1 (*WTVCFQ CPF 4CEJWDKPUMK ý999ù, YlRAC1 (*WTVCFQ GV CN. ú000ù, YlCLA4 (5\CDQ ú00ýù, YlBHM1 CPF YlBEM1 (*WTVCFQ CPF 4CEJWDKPUMK ú00úC; ú00úDù - JCXG DGGP KUQNCVGF. $[ VTCPURQUQP OWVCIGPGUKU, GKIJV VCIIGF OWVCPVU FGHGEVKXG KP J[RJCN HQTOCVKQP YGTG KFGPVKHKGF: YlSNF5, YlRAS2, YlCDC25, YlBUD6, YlGPI7, YlPPH21, CPF YlKEX2 (4KEJCTF GV CN. ú00ýù. YlMGF1 YCU KUQNCVGF D[ UKOKNCTKV[ YKVJ S. cerevisiae PHD1 QT SOK2 ()KOGPQ CPF ôKPM ý99òù CPF C. albicans EFG1, CPF UJQYP VQ JCXG VJG UCOG HWPEVKQP CU KVU JQOQNQIWGU (2oTG\-%CORQ CPF &QOsPIWG\, UWDOKVVGFù. 4GEGPVN[, YJKNG KP C UGCTEJ HQT OWVCPVU CHHGEVKPI VJG GZRTGUUKQP QH CEKFKE CPF CNMCNKPG RTQVGCUGU, UGXGTCN IGPGU CHHGEVKPI DQVJ RTQVGCUG KPFWEVKQP CPF OQTRJQIGPGUKU YGTG KFGPVKHKGF, KPENWFKPI SSY5, OPT1, MED4, CPF SIN3 ()QP\CNG\-ñQRG\ GV CN. ú00úù UGG 6CDNG ý. 7PGZRGEVGFN[, VJG XPR2 CPF YlSEC14 IGPGU EQFKPI TGURGEVKXGN[ HQT CP CNMCNKPG GZVTCEGNNWNCT RTQVGKPCUG (5\CDQ ý999ù CPF HQT C RJQURJCVKF[NKPQUKVQNíRJQURJCVKF[NEJQNKPG VTCPUHGT RTQVGKP QH VJG NCVG )QNIK (ñQRG\ GV CN. ý99òù CRRGCTGF VQ DG KPXQNXGF KP VJG FKOQTRJKE VTCPUKVKQP. 6JG YlODC IGPG EQFKPI HQT QTPKVJKPG FGECTDQZ[NCUG YCU KUQNCVGF WPFGT VJG VJGQTGVKECN RTGFKEVKQP QH KVU RWVCVKXG TQNG KP FKOQTRJKUO (ÿKOGPG\-$TGOQPV GV CN. ú00ýù. ôKPCNN[, C IGPG -YWP1- EQFKPI HQT C EGNN YCNN RTQVGKP URGEKHKE VQ VJG O[EGNKCN HQTO QH Y. lipolytica JCU CNUQ DGGP FGUETKDGF (4COQP GV CN. ý99ü; ý999ù, DWV FGNGVKQP QH VJKU IGPG FKF PQV CHHGEV OQTRJQIGPGUKU. 6JWU, WR VQ PQY OQTG VJCP ú0 IGPGU, YJKEJ CHHGEV VJG [GCUV-J[RJC VTCPUKVKQP JCXG DGGP KFGPVKHKGF (6CDNG ýù.
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6CDNG 1. )GPGU KPXQNXGF KP VJG [GCUV-J[RJC VTCPUKVKQP KP Yarrowia lipolytica 4GHGTGPEG
)GPG
(KNCOGPVCVKQP
*QOQNQIWG KP 5. EGTGXKUKCG
HOY1
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*QOQNQIWG KP %. CNDKECPU ûQ
MHY1
-
ûQ
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TUP1
RGWFQJ[RJC
TUP1
TUP1
MGF1
PF
CPH1 QT SOK2
EFG1
BEM1
-
BEM1
YF
BMH1
-
CLA4 SNF5 SIN3
RUGWFQJ[RJC
BEMH1 BMH2 CLA4 SNF5 SIN3
RAC1 STE7
TGFWEGF -
YNL180c STE7
YF HST7
RAS2 CDC25 XPR2 KEX2 PPH21 GPI7 BUD6
CDGTTCPV RUGWFQO[EGNKWO TGFWEGF
RAS2/RAS1 CDC25/SDC25 No KEX2 PPH21/PPH22 GPI7 BUD6
YF YF ûQ YF YF YF YF
(6QTTGU-)W\OCP CPF &QOKPIWG\ ý99õù (*WTVCFQ CPF 4CEJWDKPUMK ý999ù (#. &QOKPIWG\, WPRWDNKUJGFù (#. &QOKPIWG\, WPRWDNKUJGFù (*WTVCFQ CPF 4CEJWDKPUMK ú00úCù (*WTVCFQ CPF 4CEJWDKPUMK ú00úDù (5\CDQ ú00ýù (4KEJCTF GV CN. ú00ýù ()QP\CNG\-ñQRG\ GV CN. ú00úù (*WTVCFQ GV CN. ú000ù (#. &QOKPIWG\, UWDOKVVGFù (4KEJCTF GV CN. ú00ýù (4KEJCTF GV CN. ú00ýù (5\CDQ ý999ù (4KEJCTF GV CN. ú00ýù (4KEJCTF GV CN. ú00ýù (4KEJCTF GV CN. ú00ýù (4KEJCTF GV CN. ú00ýù
MED4
YF
CN.
MED4
or
YF CLA4 YF SIN3
()QP\CNG\-ñQRG\ GV ú00úù SSY5 TGFWEGF SSY5 YF ()QP\CNG\-ñQRG\ GV ú00úù OPT1 TGFWEGF OPT1 OPT1 ()QP\CNG\-ñQRG\ GV ú00úù SEC14 SEC14 SEC14 (ñQRG\ GV CN. ý99òù ODC RQN[COKPG FG- SPE1 ODC (ÿKOGPG\-$TGOQPV RGPFGPEG CN. ú00ýù YWP1 ûQ ûQ (4COQP GV CN. ý99üù #DDTGXKCVKQPU: PF, PQV FGVGTOKPGF, YF, YKVJQWV FCVC, - PQ HKNCOGPVCVKQP
CN. CN. GV
ôQWT IGPGU, HOY1, MHY1, TUP1, CPF MGF1, EQTTGURQPF VQ VTCPUETKRVKQPCN HCEVQTU, YJKEJ OC[ EQPVTQN FKTGEVN[ QT PQV OQTRJQIGPGUKU. 1VJGT NKPMU VQ VTCPUETKRVKQPCN EQPVTQNU CTG RTQXKFGF D[ YlSNF5, YJKEJ GPEQFGU C EQORQPGPV QH VJG JKIJN[ EQPUGTXGF 5ö+-5ûô EQORNGZ KPXQNXGF KP EJTQOCVKP TGOQFGNNKPI, CPF D[ YlMED4, YJKEJ GPEQFGU C UWDWPKV QH VJG OGFKCVQT EQORNGZ: DQVJ CTG JQYGXGT GZ-
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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RGEVGF VQ JCXG RNGKQVTQRKE GHHGEVU CPF OC[ PQV TGRTGUGPV RTKOCT[ VCTIGVU QH OQTRJQIGPGVKE UKIPCNU. 5GXGTCN IGPGU CTG MPQYP VQ CHHGEV EGNN RQNCTKV[ CPF CEVKP E[VQUMGNGVVQP TGCTTCPIGOGPVU KP S. cerevisiae, NKMG CDC42 CPF RAS2 (GPEQFKPI VYQ 4JQ-V[RG )62CUGU CEVKPI KP C EQORNGZ OGFKCVKPI GPXKTQPOGPVCN UKIPCNUù, CLA4 CPF BEM1 (GPEQFKPI VYQ %FEòúR DKPFKPI RTQVGKPUù, BUD6 (EQPPGEVKPI VJG CEVKP CPF VWDWNKP PGVYQTMU CV UKVGU QH CRKECN ITQYVJù. CDC25 GPEQFGU C )62 GZEJCPIG HCEVQT HQT 4CUúR CPF CEVKXCVGU VJG E#/2 RCVJYC[, YJGTGCU PPH21 GPEQFGU C V[RG ú# RTQVGKP RJQURJCVCUG TGIWNCVKPI CEVKP HKNCOGPV QTICPK\CVKQP, DQVJ DGKPI CEVKXCVGF KP TGURQPUG VQ PWVTKGPV UKIPCNNKPI. BHM1 GPEQFGU C ýò-3-3 RTQVGKP, DKPFKPI C NCTIG PWODGT QH VCTIGVU. RAC1 GPEQFGU C )62CUG QH VJG 4JQ HCOKN[, IGPGTCNN[ CEVKPI FQYPUVTGCO QH %FEòúR CPF QH VJG E#/2 RCVJYC[. 8.ý.ý ÿCVJYC[ EQPUGTXCVKQP KP Y. lipolytica, S. cerevisiae, CPF C. albicans %QORCTKUQP QH VJG IGPGU KUQNCVGF KP Y. lipolytica YKVJ VJQUG FGUETKDGF CPF EQPUGTXGF KP S. cerevisiae CPF C. albicans TCKUGU UGXGTCN KUUWGU. ôKTUV, VJTGG IGPGU -HOY1, YJKEJ GPEQFGU C RTQVGKP YKVJ C JQOGQFQOCKP JQOQNQIQWU VQ VJCV HQWPF KP VJG EJKEMGP *QZ ò.ø IGPG, MHY1, GPEQFKPI C %ú*ú-V[RG \KPE HKPIGT RTQVGKP, CPF YlRAC1, GPEQFKPI VJG HKTUV TGRQTVGF HWPICN RAC JQOQNQIWG - NCEM ENGCT JQOQNQIWGU KP S. cerevisiae CPF C. albicans. HOY1 CRRGCTU VQ UWRRTGUU UGXGTCN OQTRJQIGPGVKE FGHGEVU YJGP QXGTGZRTGUUGF CPF OC[ VJWU CEV HCKTN[ FQYPUVTGCO KP VJG RTQEGUU. +PVGTGUVKPIN[, YJGTGCU MHY1 FKUTWRVKQP NGCFU VQ EQORNGVG NQUU QH RUGWFQJ[RJCG CPF J[RJCG HQTOCVKQP, NQUU QH YlRAC1 RTGXGPVU J[RJCG HQTOCVKQP QPN[; UWRRTGUUKQP UVWFKGU KPFKECVG VJCV 4#%ý OC[ CEV FQYPUVTGCO QH /*ðý (*WTVCFQ GV CN. ú000ùù. 6YQ UGVU QH IGPGU JCXG ENGCT JQOQNQIWGU KP VJG VJTGG [GCUVU. 1PG UGV, YlTUP1, YlGMF1, YlRAS2, YlCDC25, YlCLA4 CPF YlODC, TGUWNVU KP VJG UCOG RJGPQV[RG YJGP OWVCVGF KP Y. lipolytica QT CV NGCUV KP QPG QH VJG QVJGT [GCUVU. 6JG HWPEVKQP QH VJG QVJGT ITQWR, YlSNF5, YlMED4, YlBUD6, YlGPI7, YlKEX2, CPF YlPPH21, TGOCKPU VQ DG VGUVGF KP S. cerevisiae and C. albicans. 6JG HQWT IGPGU, RAS2, CDC25, GMF1, CPF TUP1 OC[ KFGPVKH[ C E#/2-RTQVGKP-MKPCUG # RCVJYC[, EQPUGTXGF KP VJG VJTGG [GCUVU, YKVJ ðN)OHýR CPF ðN6WRýR CU FQYPUVTGCO VCTIGVU. ñKPMU VQ EGNN RQNCTKV[ CPF E[VQUMGNGVVQP TGCTTCPIGOGPVU CTG RTQXKFGF D[ CDC42, CLA4, BEM1, BHM1, BUD6, CPF PPH21. 5WTRTKUKPIN[, JQYGXGT, PGKVJGT VJG STE IGPGU QH VJG /#2þ OCVKPI RCVJYC[ PQT VJG RIM IGPGU QH VJG R* UGPUKPI RCVJYC[, YJKEJ CTG CNN KPXQNXGF KP OQTRJQIGPGUKU KP S. cerevisiae CPF KP C. albicans, UGGO VQ DG TGSWKTGF KP Y. lipolytica. ûQ GHHGEV YCU QDUGTXGF WRQP FKUTWRVKQP QH YlSTE7 (JQOQNQIWG QH ScSTE7 CPF CaHST7) QT WRQP KPVGTTWRVKQP QH CP[ QH VJG UKZ YlRIM IGPGU ()QP\CNG\-ñQRG\ GV CN. ú00úù. # UKOKNCT UKVWCVKQP YCU TGRQTVGF KP Candida Iusitaniae (ðQWPI GV CN. ú000ù CHVGT FGNGVKQP QH VJG STE12 JQOQNQIWG, UWIIGUVKPI VJCV UGXGTCN [GCUV URGEKGU OC[ PQV JCXG TGETWKVGF VJG STE RCVJYC[ HQT EQPVTQNNKPI FKOQTRJKUO.
úò0
). $CTVJ GV CN.
ôKPCNN[, C NKPM DGVYGGP PKVTQIGPQWU EQORQWPF CUUKOKNCVKQP CPF FKOQTRJKUO KU UWIIGUVGF D[ VJG DGJCXKQWT QH Y. lipolytica OWVCPVU CHHGEVGF KP SSY5 CPF OPT1, YJKEJ CTG FGNC[GF KP VJG KPFWEVKQP QH J[RJC HQTOCVKQP, CU FGUETKDGF HQT FKRNQKF UVTCKPU QH S. cerevisiae. +V VJWU CRRGCTU VJCV VJG E#/2-RTQVGKP MKPCUG # UKIPCNNKPI RCVJYC[ KU VJG OQUV EQPUGTXGF, CPF RQUUKDN[ OQTG CPEKGPV, RCVJYC[ KP FKUVCPVN[ TGNCVGF CUEQO[EGVQWU [GCUVU. &KHHGTGPV URGEKGU OC[ QT PQV JCXG TGETWKVGF CFFKVKQPCN RCVJYC[U (STE, RIMù. +P VJKU EQPVGZV, PQP-EQPXGPVKQPCN [GCUVU NKMG Y. lipolytica QHHGT KPVGTGUVKPI CNVGTPCVKXG OQFGNU VQ WPFGTUVCPF GXQNWVKQP QH VJG OQTRJQIGPGVKE RTQEGUU CPF KFGPVKH[ PGY IGPGU TGNGXCPV HQT VJG EQPVTQN QH HKNCOGPVQWU ITQYVJ KP QVJGT FKOQTRJKE HWPIK.
8.û Yarrowia lipolytica CU C OQFGN HQT OKVQEJQPFTKCN EQORNGZ + UVWF[ 6JG QDNKICVG CGTQDKE [GCUV Yarrowia lipolytica, KP EQPVTCUV VQ Saccharomyces cerevisiae, KU WPCDNG VQ IGPGTCVG VJG #62 PGGFGF HQT ITQYVJ D[ GVJCPQNKE HGTOGPVCVKQP. +P EQPUGSWGPEG, KV KU DQVJ petite PGICVKXG (KV ECP PQV UWTXKXG YKVJQWV C HWPEVKQPCN OKVQEJQPFTKCN TGURKTCVQT[ EJCKPù CPF rho PGICVKXG (KV ECP PQV UWTXKXG YKVJQWV C OKVQEJQPFTKCN IGPQOGù CPF JCU CP KPXGPVQT[ QH TGURKTCVQT[ EJCKP GP\[OGU VJCV KU SWKVG FKHHGTGPV HTQO VJG QPG KP S. cerevisiae. /KVQEJQPFTKCN RJ[UKQNQI[ CPF IGPGVKEU QH Y. lipolytica CTG QH TGNGXCPEG DQVJ HQT KVU WUG KP DKQVGEJPQNQI[, GZGORNKHKGF D[ VJG RTQFWEVKQP QH 6%# E[ENG EQORQWPFU NKMG EKVTKE CEKF CPF !MGVQINWVCTKE CEKF (#T\WOCPQX GV CN. ú000; %JGTP[CXUMC[C GV CN. ú000; +N'EJGPMQ GV CN. ú00ýù CPF KVU WUG KP DCUKE TGUGCTEJ, YJKEJ YKNN DG UWOOCTKUGF JGTG YKVJ C URGEKCN GORJCUKU QP VJG IGPGVKE CPCN[UKU QH TGURKTCVQT[ EJCKP EQORNGZ + (þGTUEJGT GV CN. ú00ýD; ú00úù. 8.û.1 üKVQEJQPFTKCN OGVCDQNKUO 1H VJG OCP[ DKQEJGOKECN RCVJYC[U VJCV VCMG RNCEG RCTVN[ QT GZENWUKXGN[ KP VJG OKVQEJQPFTKCN OCVTKZ, UGXGTCN JCXG DGGP UVWFKGF WUKPI Y. lipolytica. ñ[UKPG U[PVJGUKU KP HWPIK CPF KP Euglena QEEWTU XKC VJG !-COKPQCFKRCVG RCVJYC[, YJKEJ UVCTVU YKVJ VJG EQPFGPUCVKQP QH CEGV[N-%Q# CPF !-MGVQINWVCTCVG KPVQ JQOQEKVTCVG. Y. lipolytica YCU VJG HKTUV QTICPKUO KP YJKEJ OWVCPVU CHHGEVKPI VJKU UVGR YGTG KUQNCVGF CPF WUGF VQ ENQPG VJG LYS1 IGPG ()CKNNCTFKP CPF *GUNQV ý9õ9; 2GTG\-%CORQ GV CN. ý99üù. öJKNG LYS1 KU VJG QPN[ IGPG HQT JQOQEKVTCVG U[PVJCUG KP Y. lipolytica, VJGTG CTG VYQ KUQIGPGU (LYS20, LYS21ù KP S. cerevisiae (%JGP GV CN. ý99õDù, GZRNCKPKPI YJ[ UKPING IGPG OWVCPVU FQ PQV TGUWNV KP N[UKPG CWZQVTQRJ[ KP VJKU URGEKGU. %CVCDQNKUO QH RTQRKQP[N-%Q# (ôKI. 3ù, YJKEJ KU RTQFWEGF KP VJG RGTQZKUQOCN " QZKFCVKQP QH QFF-PWODGTGF n-CNMCPGU CPF KP VJG FGITCFCVKQP QH UGXGTCN COKPQ CEKFU, QEEWTU XKC VJG OGVJ[NEKVTKE CEKF E[ENG. $QVJ KP INWEQUG-ITQYP CPF KP n-CNMCPG-
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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(KI. 3. /KVQEJQPFTKCN OGVCDQNKUO QH RTQRKQPCVG KP ðCTTQYKC NKRQN[VKEC. ©P\[OGU QH VJG OGVJ[NEKVTKE CEKF E[ENG CTG UJCFGF KP ITG[. 4GFTCYP HTQO (7EJK[COC GV CN. ý9øúù.
úòú
). $CTVJ GV CN.
ITQYP EGNNU QH Y. lipolytica, VJG HKTUV VJTGG GP\[OGU QH VJKU E[ENG, ú-OGVJ[NEKVTCVG U[PVJCUG, ú-OGVJ[NEKVTCVG FGJ[FTCVCUG, CPF ú-OGVJ[NKUQEKVTCVG FGJ[FTCVCUG YGTG HQWPF VQ DG EQPUVKVWVKXGN[ GZRTGUUGF KP OKVQEJQPFTKC. 6JG HQWTVJ GP\[OG, úOGVJ[NEKVTCVG N[CUG YCU RTGUGPV KP DQVJ OKVQEJQPFTKC CPF E[VQRNCUO, YJGTG KV OC[ UWRRN[ VJG GPF RTQFWEV QH VJG E[ENG, R[TWXCVG, VQ XCTKQWU OGVCDQNKE RCVJYC[U (7EJK[COC GV CN. ý9øúù. 8.û.2 üKVQEJQPFTKCN TGURKTCVQT[ EJCKP 4GICTFKPI VJG OKVQEJQPFTKCN TGURKTCVQT[ EJCKP, Y. lipolytica JCU OQTG KP EQOOQP YKVJ OCOOCNU VJCP YKVJ HGTOGPVCVKXG [GCUVU NKMG S. cerevisiae. 6JG WDKSWKPQPG URGEKGU KU ó9 (þWTV\OCP ý99òù, YKVJ C PKPG WPKV KUQRTGPQKF UKFG EJCKP. 6JKU KU ENQUGT VQ ó9-ý0, CU HQWPF KP OCOOCNU VJCP VQ óü HQWPF KP S. cerevisiae. 6JG RJQURJQNKRKF EQORQUKVKQP QH OKVQEJQPFTKCN OGODTCPG HTCEVKQPU (#TVJWT CPF öCVUQP ý9õüù KU CNUQ UKOKNCT VQ OCOOCNKCP OKVQEJQPFTKC. #NN VJG GP\[OGU QH VJG OCOOCNKCP TGURKTCVQT[ EJCKP CTG RTGUGPV KP Y. lipolytica, PCOGN[ EQORNGZGU + (û#&*:WDKSWKPQPG QZKFQTGFWEVCUGù, ++ (UWEEKPCVG:WDKSWKPQPG QZKFQTGFWEVCUGù, +++ (WDKSWKPQN:E[VQEJTQOG c QZKFQTGFWEVCUGù, +8 (E[VQEJTQOG c QZKFCUGù, CPF 8 (#62 U[PVJGVCUGù. #FFKVKQPCN GP\[OGU VJCV CTG PQV HQWPF KP VJG OCOOCNKCP TGURKTCVQT[ EJCKP CTG CNVGTPCVKXG û#&* FGJ[FTQIGPCUG CPF CNVGTPCVKXG VGTOKPCN QZKFCUG. #NVGTPCVKXG û#&*:WDKSWKPQPG QZKFQTGFWEVCUGU CTG HQWPF KP OKVQEJQPFTKC HTQO RNCPVU CPF HWPIK CPF KP OCP[ DCEVGTKC (þGTUEJGT ú000ù. 6JG[ CTG UKPING UWDWPKV GP\[OGU, YJKEJ EQPVCKP QPG OQNGEWNG QH PQP-EQXCNGPVN[ DQWPF ô#&, ECTT[ QWV VJG UCOG TGFQZ TGCEVKQP CU EQORNGZ +, DWV CTG WPCDNG VQ RWOR RTQVQPU. û&*ú, VJG UKPING CNVGTPCVKXG û#&* FGJ[FTQIGPCUG QH Y. lipolytica JCU KVU CEVKXG UKVG QTKGPVGF VQYCTFU VJG GZVGTPCN HCEG QH VJG KPPGT OKVQEJQPFTKCN OGODTCPG CPF HWPEVKQPU VQ VTCPUHGT GNGEVTQPU HTQO û#&* IGPGTCVGF KP VJG E[VQRNCUO KPVQ VJG OKVQEJQPFTKCN TGURKTCVQT[ EJCKP. &GNGVKQP QH VJG NDH2 IGPG FKF PQV CRRTGEKCDN[ CHHGEV ITQYVJ TCVGU QP UGXGTCN EQORNGVG CPF OKPKOCN OGFKC (þGTUEJGT GV CN. ý999ù. 6JKU KPFKECVGU VJCV VJG CEVKXKV[ QH QVJGT GP\[OGU, NKMG IN[EGTQN-3-RJQURJCVG FGJ[FTQIGPCUG U[UVGOU CPF VJG GVJCPQN-CEGVCNFGJ[FG UJWVVNG VJCV YGTG FGUETKDGF KP S. cerevisiae (1XGTMCOR GV CN. ú000; $CMMGT GV CN. ú000ù KU UWHHKEKGPV VQ EQORGPUCVG HQT VJG NQUU QH û&*ú. 5GXGTCN CNEQJQN FGJ[FTQIGPCUG KUQGP\[OGU JCXG DGGP FGVGEVGF KP Y. lipolytica ($CTVJ CPF þWPMGN ý9õ9ù, UQOG QH YJKEJ EQWNF JCXG C HWPEVKQP KP û#&* UJWVVNKPI. #NVGTPCVKXG VGTOKPCN QZKFCUGU (#1îù CTG GZENWUKXGN[ HQWPF KP OKVQEJQPFTKC HTQO RNCPVU CPF HWPIK (ÿQUGRJ-*QTPG GV CN. ú00ýù. 6JG[ CTG CNUQ UKPING UWDWPKV GP\[OGU, YJKEJ EQPVCKP C FK-KTQP EGPVTG CPF ECTT[ QWV E[CPKFG-TGUKUVCPV GNGEVTQP VTCPUHGT HTQO WDKSWKPQN FKTGEVN[ VQ OQNGEWNCT QZ[IGP. 6JKU ÉYCUVGHWNÌ TGCEVKQP OWUV DG VKIJVN[ EQPVTQNNGF; EQPUGSWGPVN[, #1î CEVKXKV[ ECP DG FGVGEVGF KP Y. lipolytica QPN[ YJGP EGNNU CTG RQKUQPGF YKVJ KPJKDKVQTU HQT EQORNGZGU +++ QT +8. +PVGTGUVKPIN[, #1î UGGOU VQ DG GZRTGUUGF KP UVCVKQPCT[ RJCUG EWNVWTGU KP CP KPCEVKXG HQTO CPF VQ DG CEVKXCVGF D[ EQPFKVKQPU VJCV TCKUG #/2 NGXGNU (/GFGPVUGX CPF #MKOGPMQ ý999ù.
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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+P EQPVTCUV VQ HGTOGPVCVKXG [GCUV URGEKGU NKMG S. cerevisiae, YJGTG KV KU CDUGPV, VJG OWNVK-UWDWPKV RTQVQP-VTCPUNQECVKPI û#&*:WDKSWKPQPG QZKFQTGFWEVCUG MPQYP CU EQORNGZ + KU RTGUGPV KP VJG TGURKTCVQT[ EJCKP QH Y. lipolytica. ©XKFGPEG HQT VJKU JCU EQOG HTQO DKQGPGTIGVKE UVWFKGU (*GPT[ GV CN. ý9õøù, HTQO VJG FGVGEVKQP QH JKIJ OQNGEWNCT YGKIJV û#&*:HGTTKE[CPKFG QZKFQTGFWEVCUG CEVKXKV[ CHVGT UWETQUG ITCFKGPV WNVTCEGPVTKHWICVKQP ($WUEJIGU GV CN. ý99òù, CPF HTQO VJG FGVGEVKQP D[ 5QWVJGTP DNQVVKPI QH UGXGTCN û& IGPGU HQT J[FTQRJQDKE UWDWPKVU QH EQORNGZ + KP VJG OKVQEJQPFTKCN IGPQOG (ûQUGM CPF ôWMWJCTC ý99ò; þKVCPQ GV CN. ý99÷ù. S. cerevisiae ECP UWTXKXG YKVJQWV EQORNGZ + DGECWUG KV JCU PQV QPN[ VYQ GZVGTPCN (ñWVVKM GV CN. ý99øù, DWV CNUQ CP KPVGTPCN CNVGTPCVKXG û#&*:WDKSWKPQPG QZKFQTGFWEVCUG (FG 8TKGU CPF )TKXGNN ý9øø; /CTTGU GV CN. ý99ý; &G 8TKGU GV CN. ý99úù, YJKEJ JCU KVU û#&* QZKFCVKQP UKVG QTKGPVGF VQYCTFU VJG OKVQEJQPFTKCN OCVTKZ . 5KOKNCTN[, KP Y. lipolytica, C IGPGVKECNN[ GPIKPGGTGF KPVGTPCN XGTUKQP QH û&*ú YCU CDNG VQ TGUEWG VJG NGVJCNKV[ QH NQUU-QH HWPEVKQP OWVCVKQP KP EQORNGZ + (þGTUEJGT GV CN. ú00ýEù. 6JG HCEV VJCV VJG ÉPQTOCNÌ, GZVGTPCN XGTUKQP QH û&*ú ECPPQV EQORGPUCVG HQT EQORNGZ + FGHGEVU CNUQ EQPHKTOU VJCV UJWVVNKPI QH TGFQZ GSWKXCNGPV HTQO VJG OKVQEJQPFTKCN OCVTKZ KPVQ VJG E[VQUQN FQGU PQV QEEWT KP Y. lipolytica. 8.û.3 üKVQEJQPFTKCN IGPQOG # OKVQEJQPFTKCN ü.ü MD HTCIOGPV EQPVCKPKPI VJG IGPGU #62ø, #62ü, %1î3, û&ò, CPF #629 YCU ENQPGF CPF UGSWGPEGF HTQO VJG Y. lipolytica YKNF V[RG UVTCKP #6%%òòü0ý (/CVUWQMC GV CN. ý99òC; ý99òDù. 6JG UCOG ITQWR JCU CNUQ KUQNCVGF C OKVQEJQPFTKCN OWVCVKQP VJCV TGUWNVU KP TGUKUVCPEG VQ VJG EQORNGZ 8 KPJKDKVQT QNKIQO[EKP (/CVUWQMC GV CN. ý9øúù. 6JG EQORNGVG UGSWGPEG QH VJG Y. lipolytica OKVQEJQPFTKCN IGPQOG (þGTUEJGT GV CN. ú00ýCù, CU RTGUGPV KP VJG YKNF-V[RG KUQNCVG öú9 CPF VJG YKFGN[ WUGF NCD UVTCKPU ©ýú9 CPF ©ý÷0, YCU QDVCKPGF D[ EQORKNKPI EQPVKIU HTQO VJG )GPQNGXWTGU RTQLGEV (%CUCTGIQNC GV CN. ú000ù YKVJ VJG UGSWGPEGU QH UWDENQPGF HinF+++ HTCIOGPVU QH OKVQEJQPFTKCN &û# CPF ENQUWTG QH ICRU D[ FKTGEV UGSWGPEKPI QH 2%4 RTQFWEVU. #U GZRGEVGF, VJG OKVQEJQPFTKCN IGPQOG QH Y. lipolytica GPEQFGU UGXGP J[FTQRJQDKE UWDWPKVU QH EQORNGZ + (û&ý-û&ü, û&òñù, CRQE[VQEJTQOG b QH EQORNGZ +++ (%1$ù, VJTGG UWDWPKVU QH EQORNGZ +8 (%1îý, ú, 3ù CPF VJTGG UWDWPKVU QH EQORNGZ 8 (#62ü, ø, 9ù VJG 4û# EQORQPGPVU QH VJG NCTIG (rnlù CPF UOCNN (rnsù UWDWPKVU QH VJG OKVQEJQPFTKCN TKDQUQOG CPF CP KPEQORNGVG UGV QH V4û#U. 6JG UVCPFCTF HWPICN OKVQEJQPFTKCN IGPGVKE EQFG KU WUGF. 6JKU OGCPU VJCV VJG QPN[ FGXKCVKQP HTQO VJG UVCPFCTF IGPGVKE EQFG EQPEGTPU VJG 6)# EQFQP, YJKEJ KPUVGCF QH UGTXKPI CU C UVQR EQFQP KU TGCF CU VT[RVQRJCPG. 6JGTG KU PQ V4û# CDNG VQ TGCF VJG %)û EQFQP HCOKN[ GPEQFKPI CTIKPKPG. ýõ V[RG + KPVTQPU CTG RTGUGPV, ý3 QH YJKEJ EQPVCKP QRGP TGCFKPI HTCOGU HQT OCVWTCUGU VJCV HWPEVKQP KP O4û# URNKEKPI. +PVGTGUVKPIN[, UKZ QH VJG KPVTQPKE 14ôU CRRGCT VQ DG PQP-HWPEVKQPCN RUGWFQIGPGU. 6JTGG QH VJGO CTG FGHGEVKXG QPN[ DGECWUG VJG[ EQPVCKP WPTGCFCDNG %)û EQFQPU, YJKNG VJG VJTGG TGOCKPKPI JCXG CEEWOWNCVGF CFFKVKQPCN RQKPV OWVCVKQPU, HTCOGUJKHVU CPFíQT UOCNN KPUGTVKQPU.
úòò
). $CTVJ GV CN.
8.û.ý 4GURKTCVQT[ EJCKP EQORNGZ + %QORNGZ + KU C NCTIG CPF EQORNKECVGF GP\[OG (ôKI. òù, YJKEJ EQWRNGU VJG VTCPUHGT QH VYQ GNGEVTQPU HTQO û#&* VQ WDKSWKPQPG VQ VJG RWORKPI QH HQWT RTQVQPU CETQUU VJG OKVQEJQPFTKCN KPPGT OGODTCPG. +V KU CP ñ-UJCRGF OWNVK RTQVGKP EQORNGZ EQPUKUVKPI QH C OGODTCPG KPVGITCN CTO CPF C RGTKRJGTCN CTO YJKEJ RTQVTWFGU KPVQ VJG OKVQEJQPFTKCN OCVTKZ CPF EQPVCKPU VJG û#&* DKPFKPI UKVG, VJG ô/û EQHCEVQT CPF CNN MPQYP KTQP-UWNHWT ENWUVGTU. 6JG GP\[OG JCU OQTG VJCP ò0 UWDWPKVU KP OCOOCNU ((öCNMGT ý99ú; 5C\CPQX GV CN. ú000; ôGCTPNG[ GV CN. ú00ýùù CPF CV NGCUV 3÷ UWDWPKVU KP HWPIK (8KFGKTC ý99ø; &LCHCT\CFGJ GV CN. ú000ù. # ÉOKPKOCN HQTOÌ QH EQORNGZ +, CU HQWPF KP !-RTQVGQDCEVGTKC (ðCIK GV CN. ý99øù, EQPUKUVU QH ýò UWDWPKVU QPN[, CNN QH YJKEJ JCXG JQOQNQIWGU KP GWECT[QVKE EQORNGZ +, YJGTG UGXGP QH VJGUG EGPVTCN UWDWPKVU CTG GPEQFGF D[ VJG PWENGCT CPF VJG OKVQEJQPFTKCN IGPQOG, TGURGEVKXGN[ (UGG 6CDNG úù. %QORNGZ + RTQFWEGU CDQWV ò0ï QH VJG RTQVQP ITCFKGPV CETQUU VJG TGURKTCVQT[ OGODTCPG (öKMUVTQO ý9øòù CPF, UKPEG C JKIJN[ TGCEVKXG UGOKSWKPQPG KU C PCVWTCN KPVGTOGFKCVG KP KVU ECVCN[VKE E[ENG, KV KU EQPUKFGTGF C OCLQT UQWTEG QH QZ[IGP TCFKECNU RTQFWEGF KP VJG OKVQEJQPFTKCN OCVTKZ (4QDKPUQP ý99øù. %QORNGZ + KU VJG VCTIGV QH PWOGTQWU OWVCVKQPU NGCFKPI VQ OKVQEJQPFTKCN F[UHWPEVKQP KP JWOCPU (5EJCRKTC ý99ø; 5EJQP ú000; 5OGKVKPM CPF XCP FGP *GWXGN ý999ù. &GURKVG VJKU OGFKECN KORQTVCPEG, VJG TGCEVKQP OGEJCPKUO QH EQORNGZ + KU UVKNN WPMPQYP ($TCPFV ý99õ; ý999ù.
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ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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6JG KPFWEVKQP QH VJG IN[QZ[NCVG RCVJYC[ KU PGEGUUCT[ HQT VJG WVKNKUCVKQP QH CNMCPGU, HCVV[ CEKFU, CNEQJQNU, CPF CEGVCVG. +UQEKVTCVG N[CUG, VJG MG[ GP\[OG QH VJKU E[ENG, CPF KVU GPEQFKPI IGPG CTG YGNN UVWFKGF ($CTVJ CPF öGDGT ý9ø÷; ý9øõ; *QGPGU GV CN. ý99ý; þWLCW GV CN. ý99ú; /CVUWQMC GV CN. ý9ø0ù. 6JG UVTWEVWTCN IGPG JCU DGGP ENQPGF CPF UGSWGPEGF ($CTVJ CPF 5EJGWDGT ý993ù. +P EQPVTCUV VQ S. cerevisiae, VJG KUQEKVTCVG N[CUG QH Y. lipolytica KU C RGTQZKUQOCN OCVTKZ RTQVGKP CU FGOQPUVTCVGF D[ IQNF KOOWPQNCDGNKPI ($CTVJ, WPRWDNKUJGFù. *QOQNQI[ UGCTEJGU TGXGCNGF VJCV JKIJN[ EQPUGTXGF QTVJQNQIWGU VQ CNN MPQYP IGPGU KPXQNXGF KP CEGVCVG OGVCDQNKUO KP S. cerevisiae GZKUV KP VJG IGPQOG QH Y. lipolytica. &QOKPCPV OWVCVKQPU KP VJG IGPG GPR1 (IN[QZ[NCVG RCVJYC[ TGIWNCVQTù OCMG VJG EGNNU UGPUKVKXG VQ NQY EQPEGPVTCVKQPU QH CEGVCVG GXGP KP VJG RTGUGPEG QH INWEQUG (6\UEJQRRG GV CN. ý999ù. 5VWFKGU QH VJG GPR1 CNNGNGU CPF VJGKT GPEQFGF RTQVGKPU TGXGCNGF CP KPXQNXGOGPV QH VJKU RTQVGKP KP VJG OQNGEWNCT RTQEGUUGU QH CFCRVCVKQP VQ CEGVKE CEKF. )RTýR, CP KPVGITCN OGODTCPG RTQVGKP (#WIUVGKP GV CN. ú003ù, DGNQPIU VQ C PQXGN HCOKN[ QH EQPUGTXGF RTQVGKPU KP RTQ- CPF GWMCT[QVKE QTICPKUOU VJCV KU EJCTCEVGTKUGF D[ VYQ COKPQ CEKF OQVKHU (#í)ùû2#2ñ)ñ CPF 5ð)(îùôö. &GNGVKQP QH GPR1 UNQYU FQYP CFCRVCVKQP VQ CEGVKE CEKF DWV JCU PQ GHHGEV QP ITQYVJ KP VJG RTGUGPEG QH CEGVKE CEKF. ©ZRTGUUKQP QH GPR1 KU KPFWEGF D[ CEGVKE CEKF CPF OQFGTCVGN[ TGRTGUUGF D[ INWEQUG. Y. lipolytica WVKNKUGU &ñ-NCEVCVG, DWV WRVCMG QT NCEVCVG EQPXGTVKPI GP\[OGU YGTG PQV UVWFKGF UQ HCT. 8.ú.ý #EEWOWNCVKQP QH UVQTCIG ECTDQJ[FTCVGU Y. lipolytica FQGU PQV CEEWOWNCVG UVCTEJ DWV IN[EQIGP UVQTCIG QEEWTU FWTKPI GZRQPGPVKCN ITQYVJ QP INWEQUG EQPVCKPKPI OGFKWO. #EEWOWNCVKQP QH IN[EQIGP KU ECWUGF D[ C IN[EQIGP U[PVJCUG, YJKEJ KU CEVKXCVGF D[ FGRJQURJQT[NCVKQP CPF KPCEVKXCVGF D[ RJQURJQT[NCVKQP, NKMG KP S. cerevisiae (óWGKTQ\-%NCTGV GV CN. ú000ù. 8.ú.û ùVKNK\CVKQP QH J[FTQECTDQPU CU ECTDQP UQWTEG Y. lipolytica FQGU WVKNKUG n-CNMCPGU CPF ý-CNMGPGU CU ECTDQP UQWTEGU, UGG ôKIWTG 3 ($TW[P ý9÷ò; +UJKMWTC CPF ôQTUVGT ý9üý; þNWI CPF /CTMQXGV\ ý9üõ; û[PU GV CN. ý9üõù, RQN[OGVJ[NCVGF CPF EJNQTKPCVGF CNMCPGU (*CIKJCTC GV CN. ý9õõ; /WTRJ[ CPF 2GTT[ ý9øòù CU YGNN CU J[FTQZ[NCVGU CPF ENGCXGU E[ENKE J[FTQECTDQPU NKMG FKDGP\QHWTCP (4QOGT GV CN. ú00úù. # úõ M&C GZVTCEGNNWNCT GOWNUKHKGT, ECNNGF NKRQUCP, KU KPFWEGF KP EGNNU ITQYKPI QP n-CNMCPGU (%KTKINKCPQ CPF %CTOCP ý9øò; ý9ø÷ù. ñKRQUCP EQPVCKPU øøï ECTDQJ[FTCVG CPF ýúï RTQVGKP. 5OCNN FTQRNGVU QH VJG GOWNUKHKGF n-CNMCPG CFJGTG VQ VJG EGNN UWTHCEG CPF CTG KPVGTPCNKUGF CHVGTYCTFU. 7RVCMG QH n-CNMCPGU KU KPFWEKDNG CPF FWG VQ CEVKXG VTCPURQTV, DWV PQ CNMCPG URGEKHKE VTCPURQTV U[UVGO JCU DGGP EJCTCEVGTKUGF UQ HCT. 5GXGTCN CNMCPG PQP-WVKNKUKPI OWVCPVU JCXG DGGP KUQNCVGF CPF EJCTCEVGTKUGF ($CUUGN CPF
ú÷0
). $CTVJ GV CN.
/QTVKOGT ý9øú; ý9ø÷; þWLCW GV CN. ý99ú; /CWGTUDGTIGT ý99ýù. /CWGTUDGTIGT GV CN. (ý99ýù EJCTCEVGTKUGF VJTGG UWDV[RGU QH CNM# OWVCPVU, YJKEJ YGTG WPCDNG VQ WVKNKUG GKVJGT CNN V[RGU QH P-CNMCPGU (CNM#Cù, QT QPN[ UJQTV QPGU KP VJG %ø VQ %ýú TCPIG (CNM#Dù, QT QPN[ NQPI CNMCPGU KP VJG %ýü TCPIG (CNM#Eù. 6JGUG FCVC UWIIGUV VJCV Y. lipolytica OC[ EQPVCKP UGXGTCN NGPIVJ URGEKHKE CNMCPG-WRVCMG U[UVGOU KP CFFKVKQP VQ NGPIVJ URGEKHKE E[VQEJTQOG 2-ò÷0 GP\[OGU (UGG DGNQYù. #HVGT GPVT[ KPVQ VJG EGNN, n-CNMCPGU CTG J[FTQZ[NCVGF D[ C E[VQEJTQOG 2-ò÷0 OQPQQZ[IGPCUG U[UVGO. %[VQEJTQOG 2-ò÷0 YCU FGVGEVGF KP UGXGTCN UVTCKPU QH Y. lipolytica (&GNCKUUG GV CN. ý9øý; +N'EJGPMQ GV CN. ý9ø0; /CWGTUDGTIGT CPF /CVKCUJQXC ý9ø0ù. 6JG U[PVJGUKU QH VJKU GP\[OG KU KPFWEGF FWTKPI ITQYVJ QP nCNMCPGU, DWV PQV QP INWEQUG, GVJCPQN, QT CEGVCVG. &KHHGTGPEGU KP UGPUKVKXKV[ VQ INWEQUG TGRTGUUKQP FWTKPI ITQYVJ QP JGZCFGECPG CPF FGECPG KPFKECVG VJG RTGUGPEG QH UGXGTCN FKHHGTGPVN[ TGIWNCVGF E[VQEJTQOG 2-ò÷0 IGPGU KP Y. lipolytica (/CWGTUDGTIGT ý99ýù. ©KIJV FKHHGTGPV IGPGU (YlALK1 VQ YlALK8ù GPEQFKPI E[VQEJTQOGU 2ò÷0 QH VJG %ð2÷ú HCOKN[ JCXG DGGP ENQPGF CPF CPCN[UGF (+KFC GV CN. ý99ø; +KFC GV CN. ú000ù. 5KZ QH VJGUG ALK IGPGU CTG KPFWEGF D[ P-CNMCPGU CPF UVTQPIN[ TGRTGUUGF D[ IN[EGTQN DWV PQV D[ INWEQUG. YlALK1 HWPEVKQPU VQ CUUKOKNCVG n-FGECPG CPF NQPIGT OQNGEWNGU (+KFC GV CN. ý99øù. YlALK2 KU KPXQNXGF KP VJG WVKNKUCVKQP QH CNMCPGU NQPIGT VJCP n-FQFGECPG, YJGTGCU QVJGT YlALK IGPGU CTG PQV UKIPKHKECPVN[ KPXQNXGF KP VJG CUUKOKNCVKQP QH %ý0-%ýü n-CNMCPGU (+KFC GV CN. ú000ù. # PGY URGEKHKE CNMCPG-TGURQPUKXG TGIKQP (#44ýù, EQORQUGF QH VYQ FKHHGTGPV GNGOGPVU (#4©ý, #4©úù, YCU FGVGEVGF CU EKU-CEVKPI GNGOGPV KP VJG RTQOQVGT QH VJG YlALK1 IGPG (5WOKVC GV CN. ú00úù. %[VQEJTQOG 2-ò÷0 KU NQECNKUGF VQ VJG OGODTCPG QH VJG GPFQRNCUOKE TGVKEWNWO CU UJQYP D[ UWDEGNNWNCT HTCEVKQPCVKQP QH CNMCPG-ITQYP EGNNU QH Y. lipolytica (/CWGTUDGTIGT GV CN., WPRWDNKUJGFù. 1PG IGPG GPEQFKPI VJG E[VQEJTQOG TGIGPGTCVKPI û#&2-FGRGPFGPV E[VQEJTQOG 2ò÷0 TGFWEVCUG JCU CNUQ DGGP ENQPGF CPF UGSWGPEGF (6CMCIK, WPRWDNKUJGFù. 6JG RTGUGPEG QH C OGODTCPG CPEJQTKPI UGSWGPEG KP VJG û-VGTOKPCN RCTV QH VJG TGFWEVCUG CPF VJG RTQNKHGTCVKQP QH VJG OGODTCPG QH VJG GPFQRNCUOKE TGVKEWNWO ECWUGF D[ QXGTGZRTGUUKQP QH VJG TGFWEVCUG (ô{TUVGT ú00ýù KPFKECVG VJCV VJKU RTQVGKP KU NQECNKUGF VQ ©4-OGODTCPG. ý-CNMCPQN HQTOGF CHVGT VJG HKTUV UVGR KU HWTVJGT QZKFKUGF VQ VJG HCVV[ CEKF D[ C OGODTCPG-DQWPF HCVV[ CNEQJQN QZKFCUG (þGOR GV CN. ý990; /CWGTUDGTIGT GV CN. ý99úù, DWV PQV D[ û#&(2ù-FGRGPFGPV CNEQJQN FGJ[FTQIGPCUGU, YJKEJ CTG CNUQ RTGUGPV KP Y. lipolytica ($CTVJ CPF þWPMGN ý9õ9ù. 8.ú.ú *[FTQN[UKU QH HCVU /QUV UVTCKPU QH Y. lipolytica ITQY XGT[ YGNN QP HCVU CPF QKNU HTQO CPKOCNU CPF RNCPVU CPF GZRTGUU XCTKCDNG NKRQN[VKE CEVKXKV[ FGRGPFKPI QP GPXKTQPOGPVCN EQPFKVKQPU ()WGT\QPK GV CN. ú00ýù. 6JG IGPG YlLIP2 GPEQFKPI CP GZVTCEGNNWNCT NKRCUG JCU DGGP ENQPGF CPF CPCN[UGF (2KIPGFG GV CN. ú000C; ú000Dù. #V NGCUV ü QVJGT NKRCUG IGPGU JCXG TGEGPVN[ DGGP EJCTCEVGTKUGF (ÿ/ ûKECWF CPF #. &QOKPIWG\, WPRWDNKUJGFù.
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
ú÷ý
8.ú.7 (CVV[ CEKFU DKQU[PVJGUKU CPF FGITCFCVKQP ôCVV[ CEKF DKQU[PVJGUKU CPF FGITCFCVKQP YCU QPG QH VJG HKTUV RCVJYC[U KPXGUVKICVGF KP Y. lipolytica (þCOKT[Q GV CN. ý9õ9ù. #V NGCUV VYQ HCVV[ CEKF ECTTKGT U[UVGOU GZKUV KP Y. lipolytica (þQJNYGKP CPF 2CNVCWH ý9øòù, QPG DGKPI URGEKHKE HQT %ýú QT %ýò HCVV[ CEKFU, VJG QVJGT HQT UCVWTCVGF QT WPUCVWTCVGF %ýü CPF %ýø HCVV[ CEKFU. 1EVCPQKE CEKF (%øù CPF FGECPQKE CEKF (%ý0ù CTG PQV VCMGP WR D[ VJKU [GCUV. %QPVTCT[ VQ S. cerevisiae, CP KPVTCEGNNWNCT ý00 M&C HCVV[ CEKF DKPFKPI RTQVGKP KU TGCFKN[ KPFWEGF KP Y. lipolytica YJGP ITQYP QP RCNOKVCVG, CU YGNN CU C NQY OQNGEWNCT OCUU HCVV[ CEKF DKPFKPI RTQVGKP (ý÷M&Cù (&GNN'#PIGNKEC GV CN. ý99ú; ý99üù. #P CPCDQNKE CE[N-%Q# U[PVJGVCUG + CEVKXKV[ YCU HQWPF KP RGTQZKUQOGU, OKVQEJQPFTKC, CPF E[VQRNCUO CPF YCU UJQYP VQ DG TGSWKTGF HQT VJG KPEQTRQTCVKQP QH GZQIGPQWU HCVV[ CEKFU KPVQ EGNNWNCT NKRKFU. /WVCPVU FGHKEKGPV KP VJKU CEVKXKV[ JCXG DGGP KUQNCVGF (þCOKT[Q GV CN. ý9õ9ù. 6JG IGPG FAS1, GPEQFKPI VJG RGPVCHWPEVKQPCN ¾-UWDWPKV QH VJG HCVV[ CEKF U[PVJGVCUG YCU ENQPGF CPF UGSWGPEGF (þQVVKI GV CN. ý99ý; 5EJYGK\GT GV CN. ý9øøù. +VU QXGTCNN UVTWEVWTG YCU XGT[ UKOKNCT VQ VJCV QH S. cerevisiae. ôKXG IGPGU GPEQFKPI CE[N-%Q# QZKFCUGU QH FKHHGTGPV URGEKHKEKVKGU JCXG DGGP ENQPGF CPF CPCN[UGF (ñWQ GV CN. ú000; ú00ú; öCPI GV CN. ý99ø; ý999C; ý999Dù. #QZúR RTGHGTU NQPI-EJCKP CE[N-%Q#U, #QZ3 QZKFKUGU UJQTV-EJCKP CE[N-%Q#U, YJGTGCU #QZ÷R FQGU PQV UGGO VQ DG NGPIVJ URGEKHKE. 6JG HKXG KUQHQTOU QH CE[N%Q# QZKFCUG EQPUVKVWVG C òò3-M& JGVGTQRGPVCOGTKE EQORNGZ, YJKEJ KU KORQTVGF KPVQ RGTQZKUQOGU QH Y. lipolytica CU C EQHCEVQT-EQPVCKPKPI EQORNGZ (6KVQTGPMQ GV CN. ú00úù. 6JG WUG QH Y. lipolytica EGNNU HQT VJG RTQFWEVKQP QH Ä-FGECNCEVQPG HTQO CNM[N TKEKPQNGCVG (C FGTKXCVKXG QH ECUVQT QKNù JCU DGGP RCVGPVGF D[ $#5ô (&©òýúü99õ, ý993ù, (%CTFKNNQ GV CN. ý99ý; ©TEQNK GV CN. ý99úù. 6JG TQNG QH RGTQZKUQOCN ¾QZKFCVKQP CEVKXKVKGU CPF GURGEKCNN[ QH VJG CE[N-%Q# QZKFCUG KP Ä-FGECNCEVQPG CEEWOWNCVKQP JCU DGGP UVWFKGF (2CIQV GV CN. ý99ø; öCEJG GV CN. ú00ýù. 6JG CEGVQCEGV[N-%Q# VJKQNCUG GPEQFKPI IGPG YlPAT1 YCU ENQPGF D[ EQORNGOGPVCVKQP QH C OWVCPV VJCV ECP PQV WVKNKUG n-FGECPG CU UQNG ECTDQP UQWTEG. #EGVQCE[N-%Q# VJKQNCUG NQECNKUGU VQ VJG RGTQZKUQOCN OCVTKZ CPF KU GUUGPVKCN HQT WVKNKUCVKQP QH n-FGECPG (ðCOCICOK GV CN. ú00ýù. 6JG YlPOT1 IGPG GPEQFKPI C ECVCDQNKE RGTQZKUQOCN 3-QZQCE[N-%Q#-VJKQNCUG KPXQNXGF KP Ã-QZKFCVKQP QH HCVV[ CEKFU YCU ENQPGF CPF EJCTCEVGTKUGF ($GTPKPIGT GV CN. ý993ù. &KUTWRVKQP QH POT1 KPJKDKVGF VJG WVKNKUCVKQP QH QNGCVG DWV PQV VJG GNQPICVKQP QH VTKFGECPQKE CEKF VQ JKIJGT EJCKP-NGPIVJ HCVV[ CEKFU.
8.7 ÿGTQZKUQOG CUUGODN[ KP VJG [GCUV Yarrowia lipolytica +P Yarrowia lipolytica, GZVGPUKXG RGTQZKUQOG RTQNKHGTCVKQP QEEWTU FWTKPI ITQYVJ QP QNGKE CEKF (ôKI. ÷ù. 6JKU, EQODKPGF YKVJ VJG GXQNWVKQPCT[ EQPUGTXCVKQP QH VJG OGEJCPKUOU QH RGTQZKUQOG CUUGODN[ CPF VJG CXCKNCDKNKV[ QH GZEGNNGPV IGPGVKE VQQNU, OCMGU Y. lipolytica C RQYGTHWN OQFGN U[UVGO VQ UVWF[ VJG OQNGEWNCT OGEJC-
ú÷ú
). $CTVJ GV CN.
PKUOU KPXQNXGF KP RGTQZKUQOG CUUGODN[, FKXKUKQP CPF KPJGTKVCPEG (6KVQTGPMQ GV CN. ú000Dù. #P KORQTVCPV CFXCPVCIG QH Y. lipolytica, EQORCTGF VQ VJG [GCUVU Hansenula polymorpha CPF Pichia pastoris (UGG EJCRVGT õù, KP UVWF[KPI RGTQZKUQOGU KU VJCV VJGUG QTICPGNNGU CTG F[PCOKECNN[ KPVGITCVGF KP Y. lipolytica KPVQ C URGEKHKE FGXGNQROGPVCN RTQITCO, VJG FKOQTRJKE VTCPUKVKQP HTQO VJG [GCUV VQ VJG O[EGNKCN HQTO (6KVQTGPMQ GV CN. ý99õ; 6KVQTGPMQ CPF 4CEJWDKPUMK ý99øDù. $GECWUG F[UHWPEVKQP QH VJG RGTQZKUQOG ECWUGU PWOGTQWU KPJGTKVGF PGWTQNQIKECN FKUQTFGTU VJCV TGUWNV KP INQDCN FGXGNQROGPVCN FGNC[ CPF KPEQORNGVG FKHHGTGPVKCVKQP QH PGWTQDNCUVU FWTKPI RU[EJQOQVQT FGXGNQROGPV (5CEMUVGFGT CPF )QWNF ú000ù, VJG WUG QH Y. lipolytica CU C OQFGN U[UVGO YKVJ YJKEJ VQ UVWF[ VJG TQNG QH RGTQZKUQOGU KP FGXGNQROGPV CPF FKHHGTGPVKCVKQP DGEQOGU KPETGCUKPIN[ KORQTVCPV.
(KI. þ. #RRGCTCPEG QH RGTQZKUQOGU KP VJG [GCUV ðCTTQYKC NKRQN[VKEC. 6TCPUOKUUKQP GNGEVTQP OKETQITCRJU QH [GCUV EGNNU ITQYP KP OGFKWO EQPVCKPKPI GKVJGT INWEQUG QT QNGKE CEKF CU C ECTDQP UQWTEG. 2GTQZKUQOGU CTG URJGTKECN KP GNGEVTQP OKETQITCRJU, YKVJ FKCOGVGTU DGVYGGP 0.ý CPF ý.0 O. 6JGKT UK\G CPF PWODGT XCT[ FGRGPFKPI QP VJG GPXKTQPOGPVCN OKNKGW. #DDTGXKCVKQPU: /, OKVQEJQPFTKQP; û, PWENGWU; 2, RGTQZKUQOG; 8, XCEWQNG.
8.7.1 üGVCDQNKE HWPEVKQPU CPF DKQIGPGUKU QH RGTQZKUQOGU 2GTQZKUQOGU CTG QTICPGNNGU HQWPF KP OQUV GWMCT[QVKE EGNNU (5WDTCOCPK ý993ù. 6JG[ CTG FGHKPGF CU EQPVCKPKPI CV NGCUV QPG J[FTQIGP RGTQZKFG-RTQFWEKPI QZKFCUG CPF ECVCNCUG VQ FGEQORQUG VJG J[FTQIGP RGTQZKFG VQ YCVGT CPF OQNGEWNCT QZ[IGP (&G &WXG CPF $CWFJWKP ý9üüù. 2GTQZKUQOGU CTG URJGTKECN KP GNGEVTQP OKETQITCRJU, YKVJ FKCOGVGTU DGVYGGP 0.ý CPF ý.0 O. ©CEJ RGTQZKUQOG KU FGNKOKVGF D[ C UKPING WPKV OGODTCPG CPF EQPVCKPU C HKPG ITCPWNCT OCVTKZ CPF UQOGVKOGU C RCTCET[UVCNNKPG EQTG. 2GTQZKUQOGU FQ PQV EQPVCKP &û# QT CP KPFGRGPFGPV RTQVGKP U[PVJGUKU OCEJKPGT[, CU FQ OKVQEJQPFTKC CPF EJNQTQRNCUVU. #EEQTFKPIN[, CNN RGTQZ-
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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KUQOCN RTQVGKPU CTG GPEQFGF D[ PWENGCT IGPGU CPF U[PVJGUKUGF QP E[VQRNCUOKE RQN[UQOGU (2WTFWG CPF ñC\CTQY ú00ýù. 2GTQZKUQOGU UJQY TGOCTMCDNG OQTRJQNQIKECN CPF OGVCDQNKE RNCUVKEKV[; CU VJGKT PWODGT, UK\G, RTQVGKP EQORQUKVKQP, CPF DKQEJGOKECN HWPEVKQPU XCT[ FGRGPFKPI QP VJG QTICPKUO, EGNN V[RG CPFíQT GPXKTQPOGPVCN OKNKGW (XCP FGP $QUEJ GV CN. ý99úù. 6JGUG QTICPGNNGU EQORCTVOGPVCNKUG OGVCDQNKE RCVJYC[U HQT VJG FGITCFCVKQP CPF DKQU[PVJGUKU QH XCTKQWU EQORQWPFU, OQUVN[ NKRKFU. &GITCFCVKXG OGVCDQNKE RCVJYC[U KPENWFG VJG !- CPF "-QZKFCVKQP QH HCVV[ CEKFU, RWTKPG CPF COKPQ CEKF ECVCDQNKUO, CPF VJG FGITCFCVKQP QH RTQUVCINCPFKP CPF RQN[COKPGU (öCPFGTU GV CN. ú00ýù. $KQU[PVJGVKE OGVCDQNKE RCVJYC[U JQWUGF KP RGTQZKUQOGU NGCF VQ VJG HQTOCVKQP QH GVJGT RJQURJQNKRKFU (RNCUOCNQIGPUù, EJQNGUVGTQN, DKNG CEKFU, CPF RQN[WPUCVWTCVGF HCVV[ CEKFU (öCPFGTU CPF 6CIGT ý99øù. 6JG GUUGPVKCN PCVWTG QH RGTQZKUQOGU HQT PQTOCN JWOCP FGXGNQROGPV CPF RJ[UKQNQI[ KU WPFGTUEQTGF D[ VJG NGVJCNKV[ QH VJG RGTQZKUQOG DKQIGPGUKU FKUQTFGTU, KP YJKEJ RGTQZKUQOGU HCKN VQ CUUGODNG RTQRGTN[ (5CEMUVGFGT CPF )QWNF ú000ù. 6JGTGHQTG, C ITGCV FGCN QH CVVGPVKQP JCU DGGP RCKF VQ VJG OQNGEWNCT OGEJCPKUO TGIWNCVKPI RGTQZKUQOG DKQIGPGUKU. $KQIGPGUKU QH RGTQZKUQOGU, YJKEJ KPENWFGU VJGKT CUUGODN[, FKXKUKQP, CPF KPJGTKVCPEG, KU TGIWNCVGF D[ CV NGCUV ú÷ RTQVGKPU ECNNGF RGTQZKPU VJCV CTG GPEQFGF D[ VJG PEX IGPGU (6CO CPF 4CEJWDKPUMK ú00ú; 6KVQTGPMQ CPF 4CEJWDKPUMK ú00ýDù. 8.7.2 ÿGTQZKUQOG DKQIGPGUKU CU KV KU RTGUGPVGF KP EGNN-DKQNQI[ VGZVDQQMU # IGPGTCNN[ CEEGRVGF RCTCFKIO KU VJCV RGTQZKUQOGU, CU QTICPGNNGU QWVUKFG VJG UGETGVQT[ CPF GPFQE[VKE RCVJYC[U QH F[PCOKE XGUKEWNCT HNQY, EQPUVKVWVG C UVCVKE JQOQIGPQWU CPF UKPIWNCT QTICPGNNCT EQORCTVOGPV CPF VJCV RGTQZKUQOG CUUGODN[ FQGU PQV KPXQNXG KPVGTEQORCTVOGPVCN XGUKEWNCT VTCHHKEMKPI CPF OGODTCPG HWUKQP (ñQFKUJ GV CN. ý99÷ù. +P VJG ENCUUKECN RGTQZKUQOG DKQIGPGUKU OQFGN (ñC\CTQY CPF ôWLKMK ý9ø÷ù, UVTWEVWTCNN[ CPF HWPEVKQPCNN[ KFGPVKECN RGTQZKUQOGU CTG RTQRQUGF VQ GPNCTIG RTQITGUUKXGN[ D[ VJG RQUVVTCPUNCVKQPCN KORQTV QH RGTQZKUQOCN OGODTCPG CPF OCVTKZ RTQVGKPU KP DWNM CPF VJG KPEQTRQTCVKQP QH RJQURJQNKRKFU KPVQ VJG RGTQZKUQOCN OGODTCPG. 2GTQZKUQOGU VJGP WPFGTIQ FKXKUKQP VQ HQTO PGY QPGU. 8.7.3. # TGXKUKQP QH VJG RGTQZKUQOG DKQIGPGUKU RCTCFKIO: RGTQZKUQOGU CUUGODNG D[ C OWNVKUVGR RCVJYC[ 1WT TGEGPV HKPFKPIU JCXG NGCF VQ C EQORNGVG TGXKUKQP QH VJG EQPEGRV QH RGTQZKUQOGU DGKPI UVCVKE CPF JQOQIGPQWU EGNNWNCT UVTWEVWTGU (6KVQTGPMQ GV CN. ú000C; 6KVQTGPMQ CPF 4CEJWDKPUMK ý99øCù. +P Y. lipolytica, RGTQZKUQOGU, NKMG VJG )QNIK CRRCTCVWU, CEVWCNN[ EQPUVKVWVG C F[PCOKE QTICPGNNG RQRWNCVKQP (6KVQTGPMQ GV CN. ú000C; 6KVQTGPMQ CPF 4CEJWDKPUMK ú00ýDù. 6JG RQRWNCVKQP QH RGTQZKUQOGU KP C EGNN EQPUKUVU QH OCP[ UVTWEVWTCNN[ FKUVKPEV EQORCTVOGPVU (UWDHQTOUù VJCV FKHHGT KP VJGKT KORQTV EQORGVGPE[ HQT XCTKQWU RTQVGKPU CPF WPFGTIQ C URGEKHKE EQPXGTUKQP VQ OCVWTG RGTQZKUQOGU KP C VKOG-QTFGTGF OCPPGT (6KVQTGPMQ GV CN. ú000Cù. 5KZ UWD-
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HQTOU QH RGTQZKUQOGU, VGTOGF 2ý VQ 2ü, JCXG DGGP KFGPVKHKGF, RWTKHKGF, CPF EJCTCEVGTKUGF DKQEJGOKECNN[ CPF OQTRJQNQIKECNN[ (6KVQTGPMQ GV CN. ú000Cù. 6JGUG UWDHQTOU CTG QTICPKUGF KPVQ C OWNVKUVGR RGTQZKUQOG CUUGODN[ RCVJYC[ (ôKI. üù. 6JG RCVJYC[ NGCFU VQ VJG HQTOCVKQP QH OCVWTG RGTQZKUQOGU, 2ü, ECTT[KPI VJG EQORNGVG UGV QH OCVTKZ CPF OGODTCPG RTQVGKPU. 6JG GCTNKGUV KPVGTOGFKCVGU KP VJG RCVJYC[, VJG RGTQZKUQOCN XGUKENGU 2ý CPF 2ú, EQPVCKP OQUV QH RGTQZKUQOCN OGODTCPG RTQVGKPU (2/2Uù CUUQEKCVGF YKVJ OCVWTG RGTQZKUQOGU, DWV CTG CDNG VQ KORQTV QPN[ C HGY OCVTKZ RTQVGKPU (6KVQTGPMQ GV CN. ú000C; 6KVQTGPMQ CPF 4CEJWDKPUMK ú00ýCù. 2ý CPF 2ú WPFGTIQ HWUKQP VQ IGPGTCVG NCTIGT CPF FGPUGT XGUKENGU, 23 (UGG ôKIWTG üù. %QPXGTUKQP QH 23 VQ OCVWTG RGTQZKUQOGU 2ü RTQEGGFU VJTQWIJ UGXGTCN EQPUGEWVKXG UVGRU. #V GCEJ QH VJGUG UVGRU, VJG WRVCMG QH RJQURJQNKRKFU CPF UGNGEVKXG KORQTV QH C NKOKVGF UWDUGV QH OCVTKZ RTQVGKPU TGUWNV KP VJG HQTOCVKQP QH C FKUVKPEV RGTQZKUQOCN UWDHQTO VJCV KU NCTIGT CPF OQTG FGPUG VJCP KVU RTGEWTUQT (ôKI. üù. 8.7.ý ÿGTQZKUQOG HWUKQP #P KPKVKCN UVGR KP VJG OWNVKUVGR RGTQZKUQOG CUUGODN[ RCVJYC[ KPXQNXGU VJG HWUKQP QH GCTN[ KPVGTOGFKCVGU, VJG UOCNN (õ÷-ý00 POù RGTQZKUQOCN XGUKENGU 2ý CPF 2ú, VQ [KGNF ú00- VQ 3÷0-PO 23 RGTQZKUQOGU (6KVQTGPMQ GV CN. ú000Cù. 6JKU HWUKQP GXGPV JCU DGGP TGEQPUVKVWVGF in vitro, WUKPI KPVCEV RGTQZKUQOGU 2ý CPF 2ú QH ð. NKRQN[VKEC. 2GTQZKUQOG HWUKQP KU C OWNVKUVGR RTQEGUU VJCV KPENWFGU RTKOKPI, FQEMKPI, CPF HWUKQP GXGPVU (6KVQTGPMQ CPF 4CEJWDKPUMK ú000ù, UGG ôKIWTG õ. 2TKOKPI KU FTKXGP D[ #62 J[FTQN[UKU VJCV TGSWKTGU E[VQUQNKE RTQVGKPU CPF FGRGPFU QP VJG RGTQZKPU 2GZýR CPF 2GZüR (6KVQTGPMQ CPF 4CEJWDKPUMK ú000ù, VYQ #62CUGU QH VJG ### RTQVGKP HCOKN[ (#62CUGU CUUQEKCVGF YKVJ FKHHGTGPV EGNNWNCT CEVKXKVKGUù. $GHQTG RTKOKPI, 2GZýR KU CUUQEKCVGF YKVJ VJG E[VQUQNKE UWTHCEG QH 2ý, YJKNG 2GZýR CPF 2GZüR CTG DQWPF VQ VJG QWVGT UWTHCEG QH 2ú (ôKI. õù. &WTKPI RTKOKPI, #62 J[FTQN[UKU VTKIIGTU E[VQUQN-FGRGPFGPV TGNGCUG QH 2GZýR HTQO 2ý CPF QH 2GZüR HTQO 2ú, YJGTGCU 2ú-CUUQEKCVGF 2GZýR TGOCKPU DQWPF VQ VJG QTICPGNNG UWTHCEG. 2TKOKPI QH 2ý CPF 2ú RTKQT VQ VJGKT RJ[UKECN EQPVCEV EQOOKVU DQVJ HWUKQP RCTVPGTU VQ UWDUGSWGPV FQEMKPI. &QEMKPI QH RTKOGF 2ý CPF RTKOGF 2ú TGSWKTGU 2ú-CUUQEKCVGF 2GZýR CPF KU FTKXGP D[ E[VQUQN- CPF #62 J[FTQN[UKU-FGRGPFGPV TGNGCUG QH 2GZýR HTQO 2ú (ôKI. õù. &QEMKPI TGUWNVU KP VJG HQTOCVKQP QH C UVCDNG EQORNGZ (2ýí2úù CPF EQO
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(KI. ý. 2GTQZKUQOG CUUGODN[ RCVJYC[ CEVKPI KP ðCTTQYKC NKRQN[VKEC. 5KZ RGTQZKUQOCN UWDHQTOU, VGTOGF 2ý VQ 2ü, CTG QTICPKUGF KPVQ C OWNVKUVGR CUUGODN[ RCVJYC[ VJCV NGCFU VQ VJG HQTOCVKQP QH OCVWTG RGTQZKUQOGU, 2ü, ECTT[KPI VJG EQORNGVG UGV QH OCVTKZ RTQVGKPU. 5GG VGZV HQT FGVCKNU. #DDTGXKCVKQPU: #1î, CE[N-%Q# QZKFCUG; %#6, ECVCNCUG; /ñ5, OCNCVG U[PVJCUG; 6*+, VJKQNCUG; 2/2U, RGTQZKUQOCN OGODTCPG RTQVGKPU; 2GZýR, 2GZúR, 2GZüR CPF 2GZýüR, UQOG QH VJG OGODTCPG-DQWPF RGTQZKPU CUUQEKCVGF YKVJ VJG GCTNKGUV KPVGTOGFKCVGU KP VJG RCVJYC[, 2ý CPF 2ú.
OKVU 2ý CPF 2ú VQ UWDUGSWGPV HWUKQP. ôWUKQP QH 2ý CPF 2ú NGCFU HKPCNN[ VQ VJG HQTOCVKQP QH NCTIGT CPF FGPUGT RGTQZKUQOGU, 23. ôWUKQP KVUGNH KU KPFGRGPFGPV QH 2GZýR, 2GZüR, E[VQUQNKE RTQVGKPU, CPF #62 J[FTQN[UKU.
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8.7.û 6JG GPFQRNCUOKE TGVKEWNWO RNC[U CP GUUGPVKCN TQNG KP RGTQZKUQOG CUUGODN[ 6JG GCTN[ KPVGTOGFKCVGU 2ý CPF 2ú QH VJG RGTQZKUQOG CUUGODN[ RCVJYC[ CTG WPNKMGN[ VQ DG HQTOGF D[ DWFFKPI HTQO OCVWTG RGTQZKUQOGU CU RGTQZKUQOGU ECP TGCRRGCT KP [GCUV EGNNU NCEMKPI CP[ TGEQIPK\CDNG RGTQZKUQOG-NKMG UVTWEVWTGU HQNNQYKPI TGCEVKXCVKQP QT TGGZRTGUUKQP QH CP QTKIKPCNN[ FGHGEVKXG RGTQZKP-GPEQFKPI IGPG (öCVGTJCO GV CN. ý993ù. 5VWFKGU KP Y. lipolytica JCXG RTQXKFGF VJG HKTUV EQORTGJGPUKXG GXKFGPEG VJCV VJG GPFQRNCUOKE TGVKEWNWO (©4ù UGTXGU CU VJG GPFQOGODTCPG VGORNCVG HQT VJG HQTOCVKQP QH VJG GCTNKGUV RGTQZKUQOCN RTGEWTUQTU (6KVQTGPMQ GV CN. ý99õ; 6KVQTGPMQ CPF 4CEJWDKPUMK ý99øDù. 6JGUG UVWFKGU EJCNNGPIGF VJG RCTCFKIO VJCV CNN RGTQZKUQOGU CTKUG HTQO RTG-GZKUVKPI RGTQZKUQOGU CPF UWIIGUVGF C OQFGN HQT VJG HQTOCVKQP QH RTG-RGTQZKUQOCN XGUKENGU CPF VJGKT EQPXGTUKQP VQ VJG GCTN[ RGTQZKUQOCN RTGEWTUQTU 2ý CPF 2ú (6KVQTGPMQ CPF 4CEJWDKPUMK ý99øC; ú00ýC; ú00ýDù. +P VJKU OQFGN, C NKOKVGF UWDUGV QH RGTQZKUQOCN OGODTCPG RTQVGKPU
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(KI. 8. #P GUUGPVKCN TQNG HQT VJG GPFQRNCUOKE TGVKEWNWO (©4ù KP RGTQZKUQOG CUUGODN[. 6JG ©4 UGTXGU CU VJG GPFQOGODTCPG VGORNCVG HQT VJG HQTOCVKQP QH VJG GCTNKGUV RGTQZKUQOCN RTGEWTUQTU KP ðCTTQYKC NKRQN[VKEC. 5GG VGZV HQT FGVCKNU. #DDTGXKCVKQPU: 226, RTG-RGTQZKUQOCN VGORNCVG; 228ý CPF 228ú, VYQ V[RGU QH RTG-RGTQZKUQOCN XGUKENGU.
(2/2Uù KU HKTUV RQUV-VTCPUNCVKQPCNN[ VCTIGVGF VQ VJG ©4 CPF VJGP FKTGEVGF VQ C UWDFQOCKP QH VJG ©4, VJG RTG-RGTQZKUQOCN VGORNCVG (6KVQTGPMQ CPF 4CEJWDKPUMK ú00ýCù. $WFFKPI QH VJG RTG-RGTQZKUQOCN VGORNCVG NGCFU VQ VJG HQTOCVKQP QH FKUVKPEV RTG-RGTQZKUQOCN XGUKENGU, 228ý CPF 228ú (ôKI. øù. %QPXGTUKQP QH 228ý CPF 228ú VQ VJG GCTN[ RGTQZKUQOCN RTGEWTUQTU 2ý CPF 2ú KU CEEQORNKUJGF D[ WRVCMG QH FKUVKPEV UWDUGVU QH 2/2U (6KVQTGPMQ CPF 4CEJWDKPUMK ú00ýC; ú00ýDù. 8.7.ú (QNFGF, QNKIQOGTKE RTQVGKPU CTG KORQTVGF KPVQ RGTQZKUQOGU 4GEGPV HKPFKPIU KP Y. lipolytica JCXG EQPXKPEKPIN[ FGOQPUVTCVGF VJCV HQNFKPI, EQHCEVQT CVVCEJOGPV, CPF QNKIQOGTK\CVKQP QH UQOG RGTQZKUQOCN OCVTKZ RTQVGKPU QEEWT KP VJG E[VQUQN CPF CTG OCPFCVQT[ HQT VJGKT KORQTV KPVQ RGTQZKUQOGU (5\KNCTF GV CN. ý99÷; 6KVQTGPMQ GV CN. ý99ø; ú00úù. #V NGCUV VYQ FKUVKPEV RCVJYC[U HQT VJG RGTQZKUQOCN KORQTV QH HQNFGF, EQHCEVQT-EQPVCKPKPI, QNKIQOGTKE RTQVGKPU GZKUV KP Y. lipolytica. 1PG RCVJYC[ KU OGFKCVGF D[ VJG E[VQUQNKE EJCRGTQPG 2GZú0R. 2GZú0R DKPFU PGYN[ U[PVJGUKUGF OQPQOGTKE RGTQZKUQOCN OCVTKZ RTQVGKP VJKQNCUG (6JKù KP VJG E[VQUQN CPF RTQOQVGU VJG HQTOCVKQP QH C JGVGTQVGVTCOGTKE EQORNGZ EQORQUGF QH VYQ RQN[RGRVKFG EJCKPU QH GCEJ RTQVGKP (6KVQTGPMQ GV CN. ý99øù. 6JG JGVGTQVGVTCOGTKE EQORNGZ VJGP DKPFU VQ VJG OGODTCPG QH 2ú RGTQZKUQOG. 6TCPUNQECVKQP QH VJG 6JK JQOQFKOGT KPVQ VJG RGTQZKUQOCN OCVTKZ TGNGCUGU 2GZú0R OQPQOGTU DCEM VQ VJG E[VQUQN. 6JKU RCVJYC[ FQGU PQV KPXQNXG VJG 2GZ÷R-FGRGPFGPV VTCPUNQECVKQP OCEJKPGT[ VJCV CEVU CU VJG OCLQT RCVJYC[ HQT RTQVGKP KORQTV KPVQ VJG RGTQZKUQOCN OCVTKZ. # UGEQPF QNKIQOGTKE RGTQZKUQOCN OCVTKZ RTQVGKP, CE[N-%Q# QZKFCUG (#QZù, KU KORQTVGF KPVQ RGTQZKUQOGU CU C JGVGTQRGPVCOGTKE, EQHCEVQT-
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). $CTVJ GV CN.
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8.8 (WVWTG RTQURGEVU # MG[ EJCNNGPIG HQT VJG HWVWTG YKNN DG VQ KFGPVKH[ VJG OQNGEWNCT RNC[GTU KPXQNXGF KP OGODTCPG HWUKQP, UGNGEVKXG RTQVGKP KORQTV, FKXKUKQP CPF EQPXGTUKQP QH VJG FKUVKPEV RGTQZKUQOCN UWDHQTOU KFGPVKHKGF KP Y. lipolytica CPF VQ FGHKPG VJG OQNGEWNCT OGEJCPKUOU QH VJGKT CEVKQP. 6JG in vitro RGTQZKUQOG HWUKQP CUUC[ FGXGNQRGF HQT Y. lipolytica (6KVQTGPMQ GV CN. ú000Cù YKNN HCEKNKVCVG VJG KFGPVKHKECVKQP QH KPFKXKFWCN EQORQPGPVU QH VJG HWUKQP OCEJKPGT[ CPF UJQWNF RTQXKFG C ITGCVGT WPFGTUVCPFKPI QH VJG OGEJCPKUOU TGIWNCVKPI OGODTCPG HWUKQP FWTKPI DKQIGPGUKU QH VJG RGTQZKUQOG. 6JG CXCKNCDKNKV[ QH Y. lipolytica OWVCPVU VJCV CEEWOWNCVG ©4-FGTKXGF GCTN[ RTG-RGTQZKUQOCN XGUKENGU (6KVQTGPMQ GV CN. ý99õ; 6KVQTGPMQ CPF 4CEJWDKPUMK ý99øDù CPF VJG GUVCDNKUJOGPV QH C RTQEGFWTG HQT VJG RWTKHKECVKQP QH VJG GCTN[ RGTQZKUQOCN RTGEWTUQTU 2ý CPF 2ú (6KVQTGPMQ GV CN. ú000Cù HTQO VJKU [GCUV YKNN RGTOKV VJG KFGPVKHKECVKQP CPF EJCTCEVGTK\CVKQP QH RTQVGKPU CUUQEKCVGF YKVJ VJGUG FKUVKPEV XGUKENGU. 6JG WUG QH GCTN[ KPVGTOGFKCVGU KP VJG RGTQZKUQOG CUUGODN[ RCVJYC[ QRGTCVKPI KP Y. lipolytica (6KVQTGPMQ CPF 4CEJWDKPUMK ú000ù EQWNF HCEKNKVCVG VJG FGXGNQROGPV QH CP GHHKEKGPV in vitro U[UVGO YKVJ YJKEJ VQ TGEQPUVKVWVG RTQVGKP KORQTV KPVQ [GCUV RGTQZKUQOGU. 6JKU UKVWCVKQP ETGCVGU C WPKSWG QRRQTVWPKV[ VQ KFGPVKH[, WUKPI VJG RQYGTHWN CRRTQCEJ QH RTQVGQOKEU, VJG EQORQPGPVU VJCV TGIWNCVG VJG RTGEKUG F[PCOKEU QH CUUGODN[ QH RGTQZKUQOCN KORQTV OCEJKPGTKGU, CTG URGEKHKE HQT FKUVKPEV UGVU QH OCVTKZ RTQVGKPU, CPF CTG CUUQEKCVGF YKVJ FKUVKPEV RGTQZKUQOCN UWDHQTOU. #PQVJGT IQCN KU VQ FGHKPG VJG ECTIQ RTQVGKPU VJCV CTG UGNGEVKXGN[ KORQTVGF KPVQ VJG OCVTKZ QH KPFKXKFWCN RGTQZKUQOCN UWDHQTOU.
#EMPQYNGFIGOGPVU 6JG CWVJQTU YKUJ VQ VJCPM CNN VJGKT EQNNGCIWGU HTQO VJG Yarrowia EQOOWPKV[ HQT IGPGTQWUN[ UJCTKPI FCVC DGHQTG RWDNKECVKQP CPF HQT VJGKT EQPUVCPV KPVGTCEVKQP. 5RGEKCN VJCPMU CTG FWG VQ VJG OGODGTU QH VJG Yarrowia UGSWGPEKPI EQPUQTVKWO (4KEM 4CEJWDKPUMK CPF ðKG-ÿQQPI þKO KP CFFKVKQP VQ VJG CWVJQTUù CPF VQ VJG )oPQNGXWTGU ITQWR HQT UWRRQTVKPI VJG UGSWGPEKPI GHHQTV.
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4GHGTGPEGU #JNGTU 2/, )CTQHCPQ #, þGTUEJGT 5ÿ, $TCPFV 7 (ú000ù #RRNKECVKQP QH VJG QDNKICVG CGTQDKE [GCUV Yarrowia lipolytica CU C GWECT[QVKE OQFGN VQ CPCN[UG ñGKIJ U[PFTQOG OWVCVKQPU KP VJG EQORNGZ + EQTG UWDWPKVU 2556 CPF 6ðþð. $KQEJKO $KQRJ[U #EVC ýò÷9:ú÷ø-ü÷. #NDTCEJV 52 (ý993ù +PVKOCVG TGNCVKQPUJKRU QH VJG NCTIG CPF VJG UOCNN UWDWPKVU QH CNN PKEMGN J[FTQIGPCUGU YKVJ VYQ PWENGCT-GPEQFGF UWDWPKVU QH OKVQEJQPFTKCN û#&*:WDKSWKPQPG QZKFQTGFWEVCUG. $KQEJKO $KQRJ[U #EVC ýýòò:úúý-úúò #TVJWT *, öCVUQP þ (ý9õüù 6JGTOCN CFCRVCVKQP KP [GCUV:ITQYVJ VGORGTCVWTGU, OGODTCPG NKRKF, CPF E[VQEJTQOG EQORQUKVKQP QH RU[EJTQRJKNKE, OGUQRJKNKE, CPF VJGTOQRJKNKE [GCUVU. ÿ $CEVGTKQN ýúø:÷ü-üø #T\WOCPQX 6©, 5JKUJMCPQXC û8, ôKPQIGPQXC 68 (ú000ù $KQU[PVJGUKU QH EKVTKE CEKF D[ Yarrowia lipolytica TGRGCV-DCVEJ EWNVWTG QP GVJCPQN. #RRN /KETQDKQN $KQVGEJPQN ÷3:÷ú÷-÷ú9 #WIUVGKP #, $CTVJ þ, )GPVUEJ /, þQJNYGKP 5&, $CTVJ ) (ú003ù %JCTCEVGTKUCVKQP, NQECNKUCVKQP CPF HWPEVKQPCN CPCN[UKU QH )RTýR, C RTQVGKP GHHGEVKPI UGPUKVKXKV[ VQ CEGVKE CEKF KP VJG [GCUV Yarrowia lipolytica. /KETQDKQNQI[ +P RTGUU $CMMGT $/, $TQ %, þQVVGT 2, ñWVVKM /#, XCP &KLMGP ÿ2, 2TQPM ÿ6 (ú000ù 6JG OKVQEJQPFTKCN CNEQJQN FGJ[FTQIGPCUG #FJ3R KU KPXQNXGF KP C TGFQZ UJWVVNG KP 5CEEJCTQO[EGU EGTGXKUKCG. ÿ $CEVGTKQN ýøú:òõ30-òõ3õ $CTPU 5/, ñCPG &ÿ, 5QIKP /ñ, $KDGCW %, öGKUDWTI ö) (ý99ýù ©XQNWVKQPCT[ TGNCVKQPUJKRU COQPI RCVJQIGPKE %CPFKFC URGEKGU CPF TGNCVKXGU. ÿ $CEVGTKQN ýõ3:úú÷0-úú÷÷ $CTVJ ), )CKNNCTFKP % (ý99üù Yarrowia lipolytica. +P:öQNH þ (GFù ûQP-%QPXGPVKQPCN ðGCUVU KP $KQVGEJPQNQI[. # *CPFDQQM. 5RTKPIGT 8GTNCI, *GKFGNDGTI, )GTOCP[, RR 3ý3-3øø $CTVJ ), )CKNNCTFKP % (ý99õù 2J[UKQNQI[ CPF IGPGVKEU QH VJG FKOQTRJKE HWPIWU Yarrowia lipolytica. ô©/5 /KETQDKQN 4GX ý9:úý9-ú3õ $CTVJ ), þWPMGN ö (ý9õ9ù #NEQJQN FGJ[FTQIGPCUG (#&*ù KP [GCUVU. ++. û#&ì-CPF û#&2ì-FGRGPFGPV CNEQJQN FGJ[FTQIGPCUGU KP 5CEEJCTQO[EQRUKU NKRQN[VKEC. < #NNI /KMTQDKQN ý9:3øý-390 $CTVJ ), 5EJGWDGT 6 (ý993ù %NQPKPI QH VJG KUQEKVTCVG N[CUG IGPG (+%ñýù HTQO Yarrowia lipolytica CPF EJCTCEVGTK\CVKQP QH VJG FGFWEGF RTQVGKP. /QN )GP )GPGV úòý:òúú-ò30 $CTVJ ), öGDGT * (ý9ø÷ù +ORTQXGOGPV QH URQTWNCVKQP KP VJG [GCUV Yarrowia lipolytica. #PVQPKG 8CP ñGGWYGPJQGM ÷ý:ýüõ-ýõõ $CTVJ ), öGDGT * (ý9øõù )GPGVKE CPCN[UKU QH VJG IGPG +%ñý QH VJG [GCUV Yarrowia lipolytica. ðGCUV 3:ú÷÷-úüú $CUUGN ÿ, /QTVKOGT 4þ (ý9ø÷ù +FGPVKHKECVKQP QH OWVCVKQPU RTGXGPVKPI P-JGZCFGECPG WRVCMG COQPI PQP-WVKNK\KPI OWVCPVU QH Yarrowia lipolytica. %WTT )GPGV 9:÷õ9-÷øü $CUUGN ÿ$, /QTVKOGT 4þ (ý9øúù )GPGVKE CPF DKQEJGOKECN UVWFKGU QP P-CNMCPG PQP-WVKNK\KPI OWVCPVU QH Saccharomycopsis lipolytica. %WTT )GPGV ÷:õõ-øø $GTPKPIGT ), 5EJOKFVEJGP 4, %CUGN ), þPQTT #, 4CWVGPUVTCWUU þ, þWPCW ö*, 5EJYGK\GT © (ý993ù 5VTWEVWTG CPF OGVCDQNKE EQPVTQN QH VJG Yarrowia lipolytica RGTQZKUQOCN 3QZQCE[N-%Q#-VJKQNCUG IGPG. ©WT ÿ $KQEJGO úýü:ü0õ-üý3 $NC\SWG\ /#, )CPEGFQ ÿ/, )CPEGFQ % (ý99òù 7UG QH Yarrowia lipolytica JGZQMKPCUG HQT VJG SWCPVKVCVKXG FGVGTOKPCVKQP QH VTGJCNQUG ü-RJQURJCVG. ô©/5 /KETQDKQN ñGVV ýúý:úú3-úúõ
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). $CTVJ GV CN.
$QKUTCOG #, $GEMGTKEJ ÿ/, )CKNNCTFKP % (ý99üù 5NUýR, CP GPFQRNCUOKE TGVKEWNWO EQORQPGPV, KU KPXQNXGF KP VJG RTQVGKP VTCPUNQECVKQP RTQEGUU KP VJG [GCUV Yarrowia lipolytica. ÿ $KQN %JGO úõý:ýýüüø-ýýüõ÷ $QKUTCOG #, $GEMGTKEJ ÿ/, )CKNNCTFKP % (ý999ù # OWVCVKQP KP VJG UGETGVKQP RCVJYC[ QH VJG [GCUV Yarrowia lipolytica VJCV FKURNC[U U[PVJGVKE NGVJCNKV[ KP EQODKPCVKQP YKVJ C OWVCVKQP CHHGEVKPI VJG UKIPCN TGEQIPKVKQP RCTVKENG. /QN )GP )GPGV úüý:ü0ý-ü09 $QKUTCOG #, %JCUNGU /, $CDQWT #, $GEMGTKEJ ÿ/, )CKNNCTFKP % (ú00úù 5DJýR, C UWDWPKV QH VJG 5GEüý VTCPUNQEQP, KPVGTCEVU YKVJ VJG EJCRGTQPG ECNPGZKP KP VJG [GCUV Yarrowia lipolytica. ÿ %GNN 5EK ýý÷:ò9òõ-ò9÷ü $QKUTCOG #, þCDCPK /, $GEMGTKEJ ÿ/, *CTVOCPP ©, )CKNNCTFKP % (ý99øù +PVGTCEVKQP QH þCTúR CPF 5NUýR KU TGSWKTGF HQT GHHKEKGPV EQ- VTCPUNCVKQPCN VTCPUNQECVKQP QH UGETGVGF RTQVGKPU KP VJG [GCUV Yarrowia lipolytica. ÿ $KQN %JGO úõ3:30903-3090ø $QP ©, %CUCTGIQNC 5, $NCPFKP ), ñNQTGPVG $, ûGWXGINKUG %, /WPUVGTMQVVGT /, )WNFGPGT 7, /GYGU *ö, 8CP *GNFGP ÿ, &WLQP $, )CKNNCTFKP % (ú003ù /QNGEWNCT ©XQNWVKQP QH ©WMCT[QVKE )GPQOGU:*GOKCUEQO[EGVQWU ðGCUV 5RNKEGQUQOCN +PVTQPU. ûWEN #EKFU 4GU 3ý:KP RTGUU. $TCPFV 7 (ý99õù 2TQVQP-VTCPUNQECVKQP D[ OGODTCPG-DQWPF û#&*:WDKSWKPQPGQZKFQTGFWEVCUG (EQORNGZ +ù VJTQWIJ TGFQZ-ICVGF NKICPF EQPFWEVKQP. $KQEJKO $KQRJ[U #EVC ý3ýø:õ9-9ý $TCPFV 7 (ý999ù 2TQVQP VTCPUNQECVKQP KP VJG TGURKTCVQT[ EJCKP KPXQNXKPI WDKSWKPQPG--C J[RQVJGVKECN UGOKSWKPQPG UYKVEJ OGEJCPKUO HQT EQORNGZ +. $KQHCEVQTU 9:9÷-ý0ý $TQWIJVQP ÿ, 5YGPPGP &, öKNMKPUQP $/, ÿQ[GV 2, )CKNNCTFKP %, 5VKTNKPI %ÿ (ý99õù %NQPKPI QH 5©%üý JQOQNQIWGU HTQO Schizosaccharomyces pombe CPF Yarrowia lipolytica TGXGCNU VJG GZVGPV QH HWPEVKQPCN EQPUGTXCVKQP YKVJKP VJKU EQTG EQORQPGPV QH VJG ©4 VTCPUNQECVKQP OCEJKPGT[. ÿ %GNN 5EK ýý0:úõý÷-úõúõ $TW[P ÿ (ý9÷òù #P KPVGTOGFKCVG RTQFWEV KP VJG QZKFCVKQP QH JGZCFGEGPG-ý D[ Candida lipolytica. þQPKPMN. ûGF. #ECF. öGVGPUEJCR. 2TQE 5GT % ÷õ:òý-òò $WUEJIGU 4, $CJTGPDGTI ),
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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%KTKINKCPQ /%, %CTOCP )/ (ý9øòù +UQNCVKQP QH C DKQGOWNUKHKGT HTQO Candida lipolytica. #RRN ©PXKTQP /KETQDKQN òø:õòõ-õ÷0 %KTKINKCPQ /%, %CTOCP )/ (ý9ø÷ù 2WTKHKECVKQP CPF EJCTCEVGTK\CVKQP QH NKRQUCP, C DKQGOWNUKHKGT HTQO Candida lipolytica. #RRN ©PXKTQP /KETQDKQN ÷0:øòü-ø÷0 %QTFGTQ 1VGTQ 4, )CKNNCTFKP % (ý99üù ©HHKEKGPV UGNGEVKQP QH J[ITQO[EKP-$-TGUKUVCPV Yarrowia lipolytica VTCPUHQTOCPVU. #RRN /KETQDKQN $KQVGEJPQN òü:ýò3-ýòø &CTTQW\GV ©, +UUCTVGN ÿ2, ñWPCTFK ÿ, &WRWKU # (ý99øù 6JG ò9-M&C UWDWPKV QH û#&*WDKSWKPQPG QZKFQTGFWEVCUG (%QORNGZ +ù KU KPXQNXGF KP VJG DKPFKPI QH RKGTKEKFKP CPF TQVGPQPG, VYQ SWKPQPG-TGNCVGF KPJKDKVQTU. ô©$5 ñGVV ò3ý:3ò-3ø &G &WXG %, $CWFJWKP 2 (ý9üüù 2GTQZKUQOGU (OKETQDQFKGU CPF TGNCVGF RCTVKENGUù. 2J[UKQN 4GX òü:3ú3-3÷õ FG 8TKGU 5, )TKXGNN ñ# (ý9øøù 2WTKHKECVKQP CPF EJCTCEVGTK\CVKQP QH C TQVGPQPG-KPUGPUKVKXG û#&*:óü QZKFQTGFWEVCUG HTQO OKVQEJQPFTKC QH Saccharomyces cerevisiae. ©WT ÿ $KQEJGO ýõü:3õõ-3øò &G 8TKGU 5, 8CP öKV\GPDWTI 4, )TKXGNN ñ#, /CTTGU %# (ý99úù 2TKOCT[ UVTWEVWTG CPF KORQTV RCVJYC[ QH VJG TQVGPQPG-KPUGPUKVKXG û#&*- WDKSWKPQPG QZKFQTGFWEVCUG QH OKVQEJQPFTKC HTQO Saccharomyces cerevisiae. ©WT ÿ $KQEJGO ú03:÷øõ-÷9ú &GNCKUUG ÿ/, /CTVKP 2, 8GTJG[GP-$QWX[ /ô, û[PU ©ÿ (ý9øýù 5WDEGNNWNCT FKUVTKDWVKQP QH GP\[OGU KP VJG [GCUV Saccharomycopsis lipolytica, ITQYP QP P-JGZCFGECPG, YKVJ URGEKCN TGHGTGPEG VQ VJG QOGIC- J[FTQZ[NCUG. $KQEJKO $KQRJ[U #EVC üõü:õõ-90 &GNN'#PIGNKEC ©%, ©TOCEQTC /4, 5CPVQOG ÿ# (ý99üù 2WTKHKECVKQP CPF RCTVKCN EJCTCEVGTK\CVKQP QH C HCVV[ CEKF-DKPFKPI RTQVGKP HTQO VJG [GCUV, Yarrowia lipolytica. $KQEJGO /QN $KQN +PV 39:ò39-òò÷ &GNN'#PIGNKEC ©%, 5VGNNC %#, ©TOCEQTC /4, 4COQU ©*, 5CPVQOG ÿ# (ý99úù 5VWF[ QP HCVV[ CEKF DKPFKPI D[ RTQVGKPU KP [GCUV. &KUUKOKNCT TGUWNVU KP Saccharomyces cerevisiae CPF Yarrowia lipolytica. %QOR $KQEJGO 2J[UKQN $ ý0ú:úüý-úü÷ &LCHCT\CFGJ 4, þGTUEJGT 5,
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). $CTVJ GV CN.
ôNQTGU %ñ, 4QFTKIWG\ %, 2GVKV 6, )CPEGFQ % (ú000ù %CTDQJ[FTCVG CPF GPGTI[-[KGNFKPI OGVCDQNKUO KP PQP-EQPXGPVKQPCN [GCUVU. ô©/5 /KETQDKQN 4GX úò:÷0õ-÷ú9 ô{TUVGT # (ú00ýù 7PVGTUWEJWPIGP \WT JGVGTQNQIGP ©ZRTGUUKQP XQP UVGTQKFYCPFGNPFGP %[VQEJTQO 2ò÷0 5[UVGOGP KP FGT *GHG Yarrowia lipolytica. &KRNQOC VJGUKU, &TGUFGP 6GEJPKECN 7PKXGTUKV[ ôQWTPKGT 2, #DDCU #, %JCUNGU /, þWFNC $, 1IT[F\KCM &/, ðCXGT &, îWCP ÿö, 2GKVQ #, 4KDGV #/, ôG[PGTQN %, GV CN. (ý993ù %QNQECNK\CVKQP QH EGPVTQOGTKE CPF TGRNKECVKXG HWPEVKQPU QP CWVQPQOQWUN[ TGRNKECVKPI UGSWGPEGU KUQNCVGF HTQO VJG [GCUV Yarrowia lipolytica. 2TQE ûCVN #ECF 5EK 75# 90:ò9ýú-ò9ýü ôQWTPKGT 2, )CKNNCTFKP %, 2GTUW[ /#, þNQQVYKLM ÿ, XCP *GGTKMJWK\GP * (ý9øüù *GVGTQIGPGKV[ KP VJG TKDQUQOCN HCOKN[ QH VJG [GCUV Yarrowia lipolytica:IGPQOKE QTICPK\CVKQP CPF UGITGICVKQP UVWFKGU. )GPG òú:úõ3-úøú ôQWTPKGT 2, )W[CPGWZ ñ, %JCUNGU /, )CKNNCTFKP % (ý99ýù 5ECTEKV[ QH CTU UGSWGPEGU KUQNCVGF KP C OQTRJQIGPGUKU OWVCPV QH VJG [GCUV Yarrowia lipolytica. ðGCUV õ:ú÷-3ü ôTKGFTKEJ 6, 5EJGKFG & (ú000ù 6JG TGURKTCVQT[ EQORNGZ + QH DCEVGTKC, CTEJCGC CPF GWMCT[C CPF KVU OQFWNG EQOOQP YKVJ OGODTCPG-DQWPF OWNVKUWDWPKV J[FTQIGPCUGU. ô©$5 ñGVV òõ9:ý-÷ )CKNNCTFKP %, *GUNQV * (ý9õ9ù ©XKFGPEG HQT OWVCVKQPU KP VJG UVTWEVWTCN IGPG HQT JQOQEKVTCVG U[PVJCUG KP Saccharomycopsis lipolytica. /QN )GP )GPGV ýõú:ýø÷-ý9ú )KOGPQ %ÿ, ôKPM )4 (ý99òù +PFWEVKQP QH RUGWFQJ[RJCN ITQYVJ D[ QXGTGZRTGUUKQP QH PHD1, C Saccharomyces cerevisiae IGPG TGNCVGF VQ VTCPUETKRVKQPCN TGIWNCVQTU QH HWPICN FGXGNQROGPV. /QN %GNN $KQN ýò:úý00-úýýú )QP\CNG\-ñQRG\ %+, 5\CDQ 4, $NCPEJKP-4QNCPF 5, )CKNNCTFKP % (ú00úù )GPGVKE EQPVTQN QH GZVTCEGNNWNCT RTQVGCUG U[PVJGUKU KP VJG [GCUV Yarrowia lipolytica. )GPGVKEU ýü0:òýõ-òúõ )QQFYKP 6ÿ, 1TOCPF[ ÿ©, 2QWNVGT 46 (ú00ýù ñý-NKMG PQP-ñ64 TGVTQVTCPURQUQPU KP VJG [GCUV Candida albicans. %WTT )GPGV 39:ø3-9ý )WGT\QPK /©, ñCPEKQVVK 4, 8CPPKPK ñ, )CNICPQ ô, ôCXCVK ô, )CTFKPK ô, 5W\\K ) (ú00ýù 8CTKCDKNKV[ QH VJG NKRQN[VKE CEVKXKV[ KP Yarrowia lipolytica CPF KVU FGRGPFGPEG QP GPXKTQPOGPVCN EQPFKVKQPU. +PV ÿ ôQQF /KETQDKQN ü9:õ9-ø9. *CIKJCTC 6, /KUJKPC /, 6CPCMC #, ôWMWK 5 (ý9õõù 7VKNK\CVKQP QH RTKUVCPG D[ VJG [GCUV Candida lipolytica. ôCVV[ CEKF EQORQUKVKQP QH RTKUVCPG-ITQYP EGNNU. #ITKE $KQN %JGO òý:ýõò÷-ýõòø *G ô, $GEMGTKEJ ÿ/, )CKNNCTFKP % (ý99úù # OWVCPV QH õ5ñ 4û# KP Yarrowia lipolytica CHHGEVKPI VJG U[PVJGUKU QH C UGETGVGF RTQVGKP. ÿ $KQN %JGO úüõ:ý93ú-ý93õ *GPT[ /ô, 8GTJG[GP-$QWX[ /ô, û[PU ©ÿ (ý9õøù $KQIGPGUKU QH EQWRNKPI UKVG + KP VJG [GCUV Saccharomycopsis lipolytica. #TEJ +PV 2J[UKQN $KQEJKO øü:øüú-øü3. *GTTGTQ #$, ñQRG\ /%, ôGTPCPFG\-ñCIQ ñ, &QOKPIWG\ # (ý999ù Candida albicans CPF Yarrowia lipolytica CU CNVGTPCVKXG OQFGNU HQT CPCN[UKPI DWFFKPI RCVVGTPU CPF IGTO VWDG HQTOCVKQP KP FKOQTRJKE HWPIK. /KETQDKQNQI[ ýò÷:úõúõ-úõ3õ *GUNQV * (ý990ù )GPGVKEU CPF IGPGVKE GPIKPGGTKPI QH VJG KPFWUVTKCN [GCUV Yarrowia lipolytica. #FX $KQEJGO ©PI $KQVGEJPQN ò3:ò3-õ3 *KTCK /, 1JVCPK ©, 6CPCMC #, ôWMWK 5 (ý9õõù )NWEQUG-RJQURJQT[NCVKPI GP\[OGU QH Candida [GCUVU CPF VJGKT TGIWNCVKQP KP XKXQ. $KQEJKO $KQRJ[U #EVC òø0:3÷õ-3üü *QGPGU +, 5KOQP /, öGDGT *, NKRQN[VKEC. %QKNHV[ð (ý99ýù %JCTCEVGTK\CVKQP QH KUQEKVTCVG N[CUG HTQO VJG [GCUV Yarrowia lipolytica. ÿ $CUKE /KETQDKQN 3ý:ú÷ý-ú÷ø *WNN %/, ÿQJPUQP #& (ý999ù +FGPVKHKECVKQP QH C OCVKPI V[RG-NKMG NQEWU KP VJG CUGZWCN RCVJQIGPKE [GCUV Candida albicans. 5EKGPEG úø÷:ýúõý-ýúõ÷
ø ôWPEVKQPCN IGPGVKEU QH Yarrowia lipolytica
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). $CTVJ GV CN.
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9 ;GCUVU CPF HQQF URQKNCIG 5. $TWN, J. XCP FGT 8QUUGP, #. $QQTUOC, CPF (./. -NKU
#DUVTCEV (QQF RTQEGUUKPI KU EQPEGTPGF YKVh VhG RTQRGT EQPXGTUKQP QH TCY OCVGTKCNU VQ XCNWG CFFGF GPF RTQFWEVU HKV HQT EQPUWORVKQP. &WTKPI VhG RTQEGUU, OKETQDKQNQIKECN EQPÿ VCOKPCVKQP QH TCY OCVGTKCNU hCU VQ DG KPCEVKXCVGF CPF TGÿEQPVCOKPCVKQP FWTKPI RTQEGUUKPI QT WRQP RTQFWEV UVQTCIG hCU VQ DG RTGXGPVGF. +P VhKU EhCRVGT, YG UVTGUU VhG PGGF HQT CEVKXG TGUGCTEh CPF UWTXGKNNCPEG QH VhG GEQNQI[ CPF OQNGEWNCT DKQNQI[ QH HQQF URQKNCIG [GCUVU. 9G DTKGHN[ hKIhNKIhV VhG OCKP FGXGNQROGPVU KP VhG CTGC. 5WDUGSWGPVN[, VhG TGUGCTEh IGCTGF VQ KFGPVKH[KPI PGY CPVKÿ[GCUV VCTIGVU CPF VhG WPFGTUVCPFKPI QH UVTGUU TGUKUVCPEG WUKPI ENCUUKECN CRRTQCEhGU KU FKUEWUUGF. /QUV KORQTVCPV KU VhG FKUEWUUKQP QH UQTDKE CEKF TGUKUVCPEG, VhG YGCMÿQTICPKE CEKF HQQF RTGUGTXCVKXG WUGF KP VhG OCLQTKV[ QH ECUGU. 9G FKUEWUU VhG GXKFGPEG KP HCXQWT QH CP KPXQNXGOGPV QH #62 FTKXGP CEKF CPF CPKQP GZVTWUKQP RWORU KP TGUKUVCPEG FGXGNQROGPV, DWV YG CNUQ hKIhNKIhV VhG KPEQORNGVGPGUU QH VhG EWTTGPV OQFGN. /QFGTP CRRTQCEhGU WUKPI IGPQOGÿYKFG UETGGPKPI QH UVTGUU TGÿ URQPUGU QHHGT PGY VQQNU HQT CP CUUGUUOGPV QH VhG HWNN RKEVWTG QH UVTGUU TGCEVKQPU. 9G KPFKECVG GZCORNGU QH VhKU CPF FKUEWUU VhG DKQKPHQTOCVKEU OGVhQFU VhCV IQ YKVh IGPQOKEU GZRGTKOGPVU KP QTFGT VQ GZVTCEV VhG CRRTQRTKCVG MPQYNGFIG HTQO VhG FCVC. ;GCUV UVTGUU Rh[UKQNQI[, IGPGVKEU CPF ÂHWPEVKQPCN IGPQOKEU•, KU RWV KP C RGTURGEÿ VKXG QH HWVWTG TGUGCTEh CPF EhCNNGPIGU.
9.1 +PVTQFWEVKQP VQ OKETQDKCN CPF [GCUV HQQF URQKNCIG /CPWHCEVWTGF HQQFU OC[ UWHHGT UKIPKHKECPV NQUGU FWG VQ URQKNCIG D[ [GCUV ITQYVh. (QQFU KPXQNXGF KPENWFG OCTICTKPG, DGXGTCIGU, LCO, RTQEGUUGF HTWKV RTQFWEVU, CPF FTGUUKPIU þUGG G.I. &GCM 1ýý1). ;GCUVU KPXQNXGF CTG URGEKGU HTQO RTGUGTXCVKXG TGUKUVCPV UVTCKPU QH Zygosaccharomyces bailii, Z. lentus, KP CFFKVKQP VQ UVTCKPU QH Saccharomyces cerevisiae. üGPGTCNN[, UWEh [GCUVU CTG VCTIGVGF YKVh Rh[UKECN RTGUÿ GTXCVKQP VTGCVOGPVU CPF VhG CFFKVKQPU QH CPVKOKETQDKCN EQORQWPFU VQ VhG HQQF RTQFWEV QH EQPEGTP þG.I. $TWN CPF %QQVG 1ýýýû /GODTG GV CN. 1ýýýû 4QNNGT CPF %QXKNN 1ýýý). *QYGXGT, KP C YC[ SWKVG UKOKNCT VQ VhG UKVWCVKQP YKVh CPVKDKQVKEU, EQPVKPWGF WUG QH NCTIG SWCPVKVKGU QH VhG UCOG CPVKOKETQDKCN UWDUVCPEG QXGT VKOG OC[ GCUKN[ NGCF VQ VhG GOGTIGPEG QH TGUKUVCPV UVTCKPU þFKUEWUUGF GZVGPUKXGN[ KP 2KRGT GV CN. ú001). +PFGGF VhG GEQNQI[ QH [GCUVU TGNGXCPV VQ HQQF RTQEGUUKPI KU IGVVKPI OQTG EQOÿ RNGZ CU OQTG FGVCKNGF CPCN[UGU CTG DGKPI RGTHQTOGF CPF DGVVGT UGCTEhGU HQT VhG Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
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5. $TWN GV CN.
ECWUCN QTICPKUOU QH HQQF URQKNCIG CTG DGKPI ECTTKGF QWV. 6Q GPUWTG ENQUGF þCPF UWHHKEKGPV QRGP) UhGNHÿNKHG, HQQF OCPWHCEVWTGTU QHVGP KPENWFG RTGUGTXCVKXGU KP VhGKT RTQFWEVU UWEh CU YGCM QTICPKE CEKFU VQ RTGXGPV RTQFWEV URQKNCIG. *QYGXGT, VhG EQPUWOGT FGXGNQRU OQTG HGCTU HQT VhG CFFKVKQP QH CPVKOKETQDKCN EQORQWPFU VQ hKU HQQF. #U OC[ DG GZRGEVGF, VhKU ECWUGU UKIPKHKECPV RTQDNGOU HQT OKETQDKCN þQRGP) UhGNHÿNKHG þUVCDKNKV[) þG.I. üQWNF ú000). 6hGTGHQTG, VhG GXGTÿKPETGCUKPI EhCNNGPIG KP HQQF OCPWHCEVWTKPI KU VQ ICKP C RTQRGT DCNCPEG DGVYGGP RTQFWEV SWCNKV[ CPF RTQFÿ WEV UVCDKNKV[ CPF UCHGV[ þFKUEWUUGF KP øCFQ CPF ;QWUGH ú00ú). 5VTCVGIKGU VQ VhG RTQÿ VGEVKQP QH HQQF RTQFWEVU CICKPUV [GCUV ITQYVh CTG UWDLGEV VQ TGUGCTEh. 4QFTKIWGU GV CN. þú001) GZRNQTGF VhG RQUUKDKNKV[ VQ UVQR [GCUV ITQYVh D[ EQORNGVG QZ[IGP GZENWÿ UKQP. 6hG[ QDUGTXGF VhG PGGF QH Z. bailii HQT VTCEGU QH QZ[IGP VQ ITQY KP U[PVhGVKE OGFKWO. *QYGXGT, KP TKEh OGFKC, QZ[IGP KU PQV TGSWKTGF, YhKEh KPFKECVGU VhCV TGÿ UVTKEVKQP QH QZ[IGP GPVT[ KP HQQFU CPF DGXGTCIGU KU PQV C RTQOKUKPI UVTCVGI[. ÷CVWÿ TCN EQORQWPFU YKVh RTGUGTXCVKXG ECRCDKNKVKGU VhCV CRRGCN VQ VhG EWTTGPV EQPUWOGT VTGPF OKIhV KPENWFG VhG WUG QH PCVWTCNN[ QEEWTTKPI ETQR RNCPV QT CPVKOKETQDKCN EQORQWPFU FGTKXGF HTQO HQQF ITCFG OKETQQTICPKUOU þ$TWN GV CN. 1ýý7û $TWN CPF %QQVG, 1ýýýû %NGXGNCPF GV CN. ú001û øGYKU ú001û 2CVVKUQP CPF XQP *QN[ ú001û VGT 5VGGI GV CN. ú001). *QYGXGT, VhG CRRNKECVKQP QH VhGUG EQORQWPFU KU UVKNN GZVTGOGN[ NKOKVGF FWG VQ DQVh C NCEM QH WPFGTUVCPFKPI QH VhGKT OQFG QH CEVKQP CPF DGECWUG QH UVTKEV TWNGU CPF TGIWNCVKQPU HQT VhG KPVTQFWEVKQP QH PQXGN EQORQWPFU KP VhG HQQF EhCKP. +P CFFKVKQP VQ CNVGTPCVKXG CPVKOKETQDKCN EQORQWPFU, VhGTG KU CNUQ C ENGCT KPÿ VGTGUV KP VhG CRRNKECVKQP QH OKNF Rh[UKECN VTGCVOGPVU VhCV OC[ UWDUVKVWVG HQT C UGÿ XGTG hGCV VTGCVOGPV. 6hGUG KPENWFG hKIhÿRTGUUWTG RTQEGUUKPI CPF VhG CRRNKECVKQP QH RWNUGF GNGEVTKE HKGNFU þ#DGG CPF 9QWVGTU 1ýýý). *QYGXGT UKOKNCT VQ VhG CRRNKECÿ VKQP QH ÷CVWTGÿFGTKXGF CPVKOKETQDKCN U[UVGOU, VhG OQFGÿQHÿCEVKQP KU QHVGP QPN[ RCTVKCNN[ WPFGTUVQQF, CU CTG KP OCP[ ECUGU VhG DCUKE RTKPEKRNGU DGhKPF VhG GHHGEVU QH VhGUG PQXGN RTQEGUUKPI VQQNU QP HQQFU VhGOUGNXGU þFKUEWUUGF KP 5OGNV GV CN. ú00úû 9QWVGTU GV CN. ú001). (KPCNN[, KP OCP[ KPUVCPEGU HQQF OCPWHCEVWTGTU PQVKEG VhCV VhG OKETQQTICPKUOU WPFGT UVWF[ ECP GNKEKV UVTGUU TGURQPUGU CPF UWTXKXCN UVTCVGIKGU CICKPUV VhG KPFKECVGF VTGCVOGPVU. 6Q GHHGEVKXGN[ EKTEWOXGPV CPF EQODCV TGUKUVCPEG FGXGNQROGPV, KV KU PGEGUUCT[ VQ WPFGTUVCPF VhG OGEhCPKUOU VhCV IQXGTP KV þUGG G.I. #DGG CPF 9QWVÿ GTU, 1ýýýû $TWN GV CN. ú000û øCFQ CPF ;QWUUGH ú00úû 5OGNV GV CN. ú00úDû 9QWVGTU GV CN. ú00úDû &CXKFUQP CPF *CTTKUQP ú00ú). 6hG EWTTGPV FGXGNQROGPVU KP VhG HKGNF QH OQNGEWNCT OKETQDKQNQI[ KPENWFKPI IGÿ PQOGÿYKFG UETGGPKPI QH EGNNWNCT TGURQPUG CV VhG VTCPUETKRV, RTQVGKP, CPF OGVCDQNKVG NGXGN QRGP VhG RQUUKDKNKV[ HQT C HWNN CPF EQORTGhGPUKXG UVWF[ QH OKETQDKCN UVTGUU TGURQPUG CICKPUV HQQF RTGUGTXCVKQP U[UVGOU þCNUQ FKUEWUUGF KP FG 8QU ú001û $TWN GV CN. ú00ú). *GTG YG URGEKHKECNN[ HQEWU QP [GCUV. 9G UVCTV YKVh C FKUEWUUKQP QH VhG TKEh GEQNQI[ QH HQQF URQKNCIG [GCUVU, YhGTG CPF YhGP VhG[ QEEWT, CPF YhCV ECP DG FQPG VQ RTGXGPV VhGKT ITQYVh. 5WDUGSWGPVN[, YG YKNN UhQY VhG FKXGTUKV[ QH CPVKÿ [GCUV VCTIGVU FGTKXGF HTQO IGPGVKE UVWFKGU KP VhG RCUV CPF OQTG TGEGPVN[ HTQO PQXGN IGPQOGÿYKFG UVWFKGU. 9G FKUEWUU URGEKHKE UVTGUU TGUKUVCPEG OGEhCPKUOU VhCV [GCUV EGNNU hCXG UVCTVKPI YKVh VhGKT IGPGVKE DCEMITQWPF þTGUKUVCPEG RTQXKFKPI IGPGU ö IGPG RTQFWEVU). (KPCNN[, YG YKNN KNNWUVTCVG OQFGTP DKQKPHQTOCVKEU VQQNU CPF CPCN[ÿ UGU CRRNKGF KP VhKU TGUGCTEh CTGC þUGG CNUQ $WUUGOCMGT GV CN. ú001).
ý ;GCUVU CPF HQQF URQKNCIG
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9.2 6JG GEQNQI[ QH HQQF URQKNCIG [GCUVU 9.2.1 &KXGTUKV[ QH [GCUVU (QQF URQKNCIG [GCUVU CTG HQWPF KP OCP[ GPXKTQPOGPVU. ÷QVKEGCDN[ RTQFWEGF HQQF RTQFWEVU VhCV CTG OKNFN[ CEKFKE þUGG RCTCITCRh 1) CPF EQPVCKP RTGUGTXCVKXGU, C UKIÿ PKHKECPV COQWPV QH ECTDQP, CPF UWHHKEKGPV PKVTQIGP CTG MPQYP hCDKVCVU HQT UWEh [GCUVU. 5RGEKGU GPEQWPVGTGF KPENWFG VhG Zygosaccharomyces bailii þ&GCM 1ýý1) CPF Zygosaccharomyces lentus þ5VGGNU GV CN. 1ýýý). # TGEGPV TGRQTV D[ 5VTCVHQTF GV CN. þú00ú) FGUETKDGU [GV CPQVhGT PGY URGEKGU KPXQNXGF KP HQQF URQKNCIG Candida davenporti PCOGF CHVGT VhG NCVG RTQHGUUQT &CXGPRQTV, CP GZRGTV KP VhG GEQNQI[ QH HQQF URQKNCIG [GCUVU. 6hKU [GCUV KU GZVTGOGN[ CEKF TGUKUVCPV WR VQ R* 1.ù CPF OQFÿ GTCVGN[ RTGUGTXCVKXG TGUKUVCPV. +V YCU VTCEGF DCEM VQ URGEKGU MPQYP VQ EQNQPK\G YCURU CPF KV KU h[RQVhGUKUGF VhCV VhTQWIh VhKU GEQNQIKECN PKEhG KV YCU CDNG VQ KPHGEV UQHV FTKPM RTQFWEVU. +P CFFKVKQP, Saccharomycs cerevisiae CPF Candida lipolytica CTG UQOGVKOGU CDNG VQ VQNGTCVG CEKFKE GPXKTQPOGPVU CPF FKURNC[ TGUKUVCPEG VQ YGCM CEKF RTGUGTXCÿ VKXGU þFG $QGT CPF ÷KGNUGP 1ýýõ, $CKTFÿ2CTMGT GV CN. 1ýýô). (KPCNN[, Zygosaccharomyces rouxii KU CP KORQTVCPV NQY R* VQNGTCPV URQKNCIG [GCUV VhCV KU KPXQNXGF KP hKIh UWICT EQPVCKPKPI HQQF RTQFWEVU UWEh CU HQPFCPV, EhQEQNCVG, CPF UQ[ UCWEG DGECWUG QH KVU CDKNKV[ VQ ITQY CV XGT[ NQY YCVGT CEVKXKVKGU. øKMG QVhGT HGTOGPVCVKXG [GCUVU, KV FQGU RTQFWEG ICU þøGICP CPF 8Q[UGV 1ýý1). (WTVhGTOQTG, VhG QTICPKUO KU CDNG VQ ITQY CV C YKFG R* TCPIG HTQO R* 1.ô WR VQ ô.0 þ6QMWQMC 1ýý3). +P OGÿ FKWO EQPVCKPKPI EKVTKE CEKF CV R* ú.õ, C 30ó TGFWEVKQP KP ITQYVh YCU QDUGTXGF EQORCTGF VQ VhG QRVKOWO R* 3.õ VQ õ þ/GODTo GV CN. 1ýýý). 6hG[ CNUQ UhQYGF C NKPGCT ITQYVh TGFWEVKQP YKVh CP KPETGCUGF UWICT EQPEGPVTCVKQP WR VQ 70ó. (WTVhGT PKEhGU QH HQQF URQKNCIG [GCUVU KPENWFG ECTECUUGU QH CPKOCNU KP QT QP YhKEh VhG[ OC[ VhTKXG þ*KPVQP GV CN ú00ú). #TEVKE GPXKTQPOGPVU YGTG CPVKEKRCVGF VQ DG VhG UQWTEG QH Mrakia URR. UVTCKPU VhCV CTG KPXQNXGF KP URQKNCIG QH HTWKV LWKEG UVQTGF CV NQY VGORGTCVWTG þJ./.$./. XCP FGT 8QUUGP GP $.J. *CTVQI, WPRWDNKUhGF FCVC). 9.2.2 )TQYVJ EJCTCEVGTKUVKEU ;GCUV URQKNCIG KU EhCTCEVGTKUGF D[ XKUKDNG ITQYVh QP VhG UWTHCEG QH RTQFWEVU þYhKVG VQ ETGCO EQNQWTGF RCVEhGU) QT D[ ICU RTQFWEVKQP KPENWFKPI DWDDNKPI UQHV FTKPMU, LCOU, CPF GZRCPUKQP QH HNGZKDNG RCEMCIKPI. +P HCEV, [GCUVU YKNN RTQNKHGTCVG KP OCP[ HQQF RTQFWEVU YhGP RTGUGPV. *QYGXGT, QHVGP VhG[ YKNN DG RTGUGPV CU C OKPQTKV[ COQPI DCEVGTKC KP URQKNGF HQQF RTQFWEVU. òPN[ KP VhQUG RTQFWEVU YhGTG ITQYVh QH DCEVGTKC KU UWRRTGUUGF D[ VhG RTGXCKNKPI EQPFKVKQPU, [GCUVU OC[ hCXG C UGNGEVKXG CFXCPVCIG CPF CTG PQV QWV RTGHQTOGF D[ VhG TCRKF ITQYVh QH DCEVGTKC WPFGT OKNF GPXKTQPOGPVCN EQPFKVKQPU. #NYC\GGT GV CN. þú00ú) UhQYGF VhCV Lactobacillus plantarum CPF S. cerevisiae, YhKEh CTG DQVh YGCM CEKF VQNGTCPV OKETQQTICPKUOU, CTG CDNG VQ URQKN EKVTWU LWKEGU DGHQTG CPF CHVGT RCUVGWTKUCVKQP. +V CRRGCTGF HTQO VhGKT UVWF[ VhCV VhG OQTG VGORGTCVWTG UGPUKVKXG S. cerevisiae UhQYU C HCUVGT ITQYVh TGÿ
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5. $TWN GV CN.
EQXGT[ CHVGT RCUVGWTKUCVKQP VhCP L. plantarum, YhKEh KU KP C PQPÿKPLWTGF UKVWCVKQP OQTG VGORGTCVWTG TGUKUVCPV. #FTKGPPG GV CN. þú00ú) CPCN[UGF VhG hGCV TGUKUVCPEG QH [GCUVU, OQWNFU, CPF NCEVQDCEKNNK QDVCKPGF HTQO URQKNGF CEKF HQQF RTQFWEVU. S. cerevisiae YCU VhG OQUV hGCV TGUKUVCPV QTICPKUO KP EKVTCVG DWHHGT. 6hG hGCV TGUKUVCPEG CPF VhWU VhG RTQEGUU UWTXKXCN ECRCEKV[ QH VhKU [GCUV YCU HWTVhGT GNGXCVGF KP HTWKV LWKEG GPXKTQPOGPVU. 9.2.3 )TQYVJ RJ[UKQNQI[ 6[RKECN UGNGEVKXG EQPFKVKQPU HQT [GCUV RTQNKHGTCVKQP KPUVGCF QH DCEVGTKCN ITQYVh CTG NQY R*, NQY YCVGT CEVKXKV[, NQY VGORGTCVWTGU, CPF CEKF RTGUGTXCVKXGU. &WTKPI FGÿ EQORQUKVKQP QH RNCPV OCVGTKCNU CPF QVhGT QTICPKE OCVGTKCNU, hKIh EQPEGPVTCVKQPU QH CEGVCVG CPF NCEVCVG CTG VhG RTQFWEVU QH DCEVGTKCN HGTOGPVCVKQP. ;GCUVU HQTO C RCTV QH OCP[ QH VhGUG GEQU[UVGOU CPF VhWU OWUV CV NGCUV UQOGVKOGU DG SWKVG TGUKUVCPV VQ VhG RTGUGPEG QH YGCM QTICPKE CEKFU. 6hKU UKVWCVKQP KU EhCNNGPIKPI VhG HQQF KPFWUVT[ DGECWUG KV NGCFU VQ CP KPETGCUG KP TGUKUVCPEG VQ VhG OCLQT QTICPKE CEKF HQQF RTGÿ UGTXCVKXGU. Z. bailii, Zygosaccharomyces bisporus, CPF Z. lentus CTG VhG OCKP CEKFÿ VQNGTCPV URGEKGU. /QNGEWNCT V[RKPI VGEhPKSWGU CU FGUETKDGF D[ %QWVQ GV CN. 1ýýñ CPF XCP FGT 8QUUGP GV CN. ú003) UhQYGF VhCV VhGUG Zygosaccharomyces URGEKGU CTG VhG OCLQT URQKNCIG QTICPKUOU KP OC[QPPCKUGU CPF NQY R* VQOCVQ UCWEGU. Z. bailii KU CNUQ CP KORQTVCPV URQKNCIG CIGPV QH YKPG FWG VQ KVU hKIh GVhCPQN VQNGTCPEG þ-CNCVhGPQU GV CN. 1ýýõ). (WTVhGTOQTG, VhKU QTICPKUO FQGU OGVCDQNKUG CEGVKE CEKF KP VhG RTGUGPEG QH EQORNGZ OKZVWTGU QH UWICTU KP YKPG HGTOGPVCVKQPU þ5QWUC GV CN. 1ýýñû 1ýýô). Z. bailii KU CDNG VQ ECVCN[UG QZKFCVKXG FGITCFCVKQP QH YGCM CEKF RTGUGTXCVKXGU UWEh CU DGP\QKE CEKF CPF UQTDKE CEKF YhGTG QVhGT URGEKGU UWEh CU S. cerevisiae NKOKV VhG GPVT[ QH CEKF KPVQ VhG EGNNU þ2KRGT GV CN. ú001 UGG CNUQ FG ÷QDGN GV CN. ú001). +PVTQFWEVKQP CPF GZRTGUUKQP QH C UKPING IGPG HTQO Z. bailii CNNQYU S. cerevisiae EGNNU VQ ITQY QP DGP\QCVG CPF UQTDCVG þ/QNNCRQWT CPF 2KRGT ú001). Z. bailii EGNNU FQ DGhCXG FKHHGTGPVN[ HTQO S. cerevisiae WPFGT R* UVTGUU. %hGPI GV CN þ1ýýý) UhQYGF VhCV VTGhCNQUG, C OCLQT UVTGUU RTQVGEVCPV FQGU CEEWOWNCVG TCRKFN[ KP S. cerevisiae GZRQUGF VQ NQY R* YhGTGCU VhKU TGURQPUG KU PQV QDUGTXGF KP VhG OQTG RTGÿ UGTXCVKXG TGUKUVCPV Z. bailii EGNNU. 9.2.4 ÿTGUGTXCVKQP UVTCVGIKGU 6hG WUG QH OQNGEWNCT V[RKPI VGEhPKSWGU HQT VTCEKPI UQWTEGU QH [GCUV EQPVCOKPCVKQP UhQYGF VhCV YKPG KU UQOGVKOGU KPXQNXGF CU VhG UQWTEG QH Z. bailii URQKNCIG KP YKPG DCUGU UCWEGU þXCP FGT 8QUUGP, WPRWDNKUhGF QDUGTXCVKQP). 6hGUG VGEhPKSWGU HWTVhGT UhQYGF VhCV Z. bailii CPF Z. bisporus OC[ GUECRG HTQO ENGCPKPI KP RNCEG þ%+2) TGÿ IKOGU KP VhG HQQF KPFWUVT[ þXCP FGT 8QUUGP GV CN. ú003). /QUV QHVGP, VhKU KU VhG EQPUGSWGPEG QH RNCEGU KP VhG RTQEGUU GSWKROGPV VhCV EQPVCKP URCEGU, YhKEh CTG PQV TGCEhGF D[ VhG %+2. 6hG OCKP ECWUGU QH [GCUV URQKNCIG KP VhG HQQF KPFWUVT[ ECP DG FKTGEVGF VQYCTFU TGUKUVCPEG FGXGNQROGPV KP CP GPXKTQPOGPV YhGTG [GCUVU CTG EQPÿ
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UVCPVN[ VTKIIGTGF D[ UWDÿNGVhCN EQPEGPVTCVKQPU QH RTGUGTXCVKXGU. (WTVhGT ECWUGU CTG C NCEM QH h[IKGPG, RQWT SWCNKV[ QH TCY OCVGTKCNU, CPF HCKNKPI h[IKGPKE FGUKIP QH RTQEGUU GSWKROGPV. 6hG WUG QH RTGUGTXCVKQP UVTCVGIKGU ECP PGXGT TGRNCEG IQQF OCPWHCEVWTKPI RTQEGUUGU DWV ECP RTQXKFG CFFKVKQPCN RTQVGEVKQP. 5KPEG YGCM CEKF RTGUGTXCVKXGU CTG PQV HWNN[ TGNKCDNG, VhGTG KU EGTVCKPN[ VhG PGGF HQT CFFKVKQPCN OGCUÿ WTGU FKTGEVGF VQYCTFU CRRTQRTKCVG VCTIGV UKVGU KP URQKNCIG [GCUVU. +P QTFGT VQ KFGPVKH[ UWKVCDNG VCTIGVU HQT PQXGN RTGUGTXCVKQP UVTCVGIKGU QT UVTCVGÿ IKGU VhCV GPhCPEG VhG GHHGEVKXGPGUU QH EWTTGPV RTGUGTXCVKXGU C OGEhCPKUVKE WPFGTÿ UVCPFKPI QH VhG HWPEVKQPKPI QH [GCUV EGNNU KU PGGFGF. 5KPEG VCTIGVU PGGF VQ DG EhCTÿ CEVGTKUVKE CPF GUUGPVKCN HQT [GCUV EGNNU, VhG RTKOCT[ ECPFKFCVGU CTG NKMGN[ VQ GOCPCVG HTQO MPQYNGFIG QH VhG UVTWEVWTG CPF HWPEVKQP QH VhG [GCUV GPXGNQRG.
9.3 6JG [GCUV GPXGNQRG, CPVKHWPICN VCTIGVU, CPF (RTGUGTXCVKQPþ UVTGUU TGUKUVCPEG +P QTFGT VQ WPFGTUVCPF [GCUV UVTGUU TGUKUVCPEG, KV KU CRRTQRTKCVG VQ EQPEKUGN[ FKUEWUU VhG QTICPK\CVKQP QH VhG [GCUV EGNN GPXGNQRG þEGNN OGODTCPG CPF EGNN YCNN). #U C OQFGN U[UVGO, YG FKUEWUU VhG IGPGVKECNN[ DGUV EhCTCEVGTKUGF DCMGTU• [GCUV Saccharomyces cerevisiae. +VU CRRTQRTKCVGPGUU CU C TGHGTGPEG U[UVGO HQT HQQF URQKNCIG [GCUV YCU CTIWGF GZVGPUKXGN[ DGHQTG þ$TWN GV CN. 1ýý7û -NKU GV CN. ú00ú CPF TGHGTÿ GPEGU VhGTGKP) GXGP VhQWIh VhGTG CTG UKIPKHKECPV FKHHGTGPEGU KP YGCM CEKF TGUKUVCPEG OGEhCPKUOU CU YG PQY MPQY þ/QNNCRQWT CPF 2KRGT ú00ú). (TQO VhKU RQKPV QP TGÿ HGTTKPI VQ Â[GCUV• YKNN KORN[ TGHGTTKPI VQ VhG UKVWCVKQP KP S. cerevisae WPNGUU URGEKÿ HKGF QVhGTYKUG. (QT CP GZVGPUKXG CEEQWPV QH VhG EQORQUKVKQP CPF VhG F[PCOKEU QH VhG [GCUV YCNN UGG -NKU GV CN. þú00ú). (KIWTG 1 UWOOCTKUGU EWTTGPV XKGYU QP RQUUKÿ DNG CPVKÿ[GCUV VCTIGVU RTGUGPV KP VhG EGNN GPXGNQRG. +PFKXKFWCN RTQEGUUGU CPF RTQÿ VGKPU YKNN DG FKUEWUUGF KP VhG EQWTUG QH VhG RCTCITCRhU. 6CDNG 1 RTQXKFGU KPHQTOCÿ VKQP QP VhG V[RGU QH OQNGEWNGU IGPGTCNN[ HQWPF KP VhG [GCUV EGNN GPXGNQRG. 6hG GPXGNQRG EQPVCKPU RhQURhQNKRKFU UKOKNCT VQ QVhGT OGODTCPGU, URGEKHKE UVGTQN EQORQPGPVU EhCTCEVGTKUVKE HQT HWPIK CPF RNCUOCOGODTCPGRTQVGKPU þG.I. OGODTCPG NQECNKUGF VTCPURQTVGTU), CPF GUUGPVKCN EGNN YCNN EQORQWPFU þXCP FGT 4GUV 1ýýõû -NKU GV CN. ú00ú). *GTG, YG YKNN OCKPN[ FKUEWUU VhG QEEWTTGPEG KP CPF VhG TQNG VhCV RTQVGKPU RNC[ KP VhG UVTWEVWTKPI QH VhG EGNN GPXGNQRG. (QT C FKUEWUUKQP QH VhG NKRKF CPF UVGTQN EQORQUKVKQP QH VhG RNCUOC OGODTCPG CPF KVU EQPUGSWGPEGU HQT VhG FGXGNQROGPV QH CPVKHWPICN UVTCVGIKGU UGG G.I. /QTUEhhCWUGT þú00ú), 5CPINCTF CPF òFFU þú00ú). +P QWT FKUEWUUKQP QH CPVKHWPICN VCTIGVU, YG YKNN HTGSWGPVN[ FTCY HTQO GZCORNGU HTQO OGFKECNN[ QTKGPVGF TGUGCTEh. *KIhN[ CDWPFCPV EGNN OGODTCPG RTQVGKPU CTG VhQUG KPXQNXGF KP VhG EQPUVTWEVKQP QH VhG EGNN YCNN þK.G. EhKVKP CPF INWECP U[PVhCUGU UGG G.I. -NKU GV CN. ú00ú). (WTVhGTÿ OQTG, VhGTG CTG WDKSWKVQWU RTQVGKPU KPXQNXGF KP VhG VTCPURQTV QH NQY OQNGEWNCT YGKIhV EQORQWPFU þK.G. hGZQUG VTCPURQTV CPF GZVTWUKQP QH HQTGKIP EQORQWPFU UGG G.I. -TWEMGDGTI GV CN. 1ýýô CPF ú001) CPF HKPCNN[ VhG hKIhN[ CDWPFCPV RNCUÿ OCOGODTCPG NQECNKUGF RTQVQP RWORKPI #62ÿCUG þ/QTUQOOG GV CN. ú000).
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5. $TWN GV CN.
ÿ+ü-RTQVGKPU NKPMGF VQ DGVC-1,3 INWECP PGVYQTM > 40 KPHGTTGF )ÿ+-DGVC1,ý INWECP NKPMGF OCPPQRTQVGKPU
DGVC-1,6 INWECP
ÿWVCVKXG EGNN YCNN ETQUU-NKPMKPI GP\[OGU G.I. )CU1R û úTG9R
DGVC-1,3 INWECP EJKVKP
EJKVKP U[PVJGUKU (EJKVKP U[PVJCUG 3)
INWECP U[PVJGUKU (INWECP U[PVJCUG 1,2)
JGZQUG WRVCMG (DGPFGF: UVTGUU EQPFKVKQPU) RTGUGTXCVKXG GZVTWUKQP (RNGKQVTQRKE FTWI TGUKUVCPEG RWOR ÿFT12)
EGNN YCNN UVTGUU UGPUQTU 9UE1, /KF2
RTGUWOGF NKRQRJ[NKE YGCM-CEKF UVTGUU UGPUQT
CEKF GZVTWUKQP RTQVQP-RWORKPI ÿOC1
(KI. 1. 6hG EGNN GPXGNQRG QH VhG [GCUV Saccharomyces cerervisiae KU QHVGP WUGF CU VhG OQÿ NGEWNCT OQFGN HQT OCP[ HQQF URQKNCIG HWPIK þG.I. $TWN GV CN. 1ýý7û FG ÷QDGN GV CN. ú001). 6hG EGNNU hCXG VhG CDKNKV[ VQ CFLWUV VQ GPXKTQPOGPVCN UVTGUU CRRNKGF YKVh VhG RTQFWEVKQP QH URGEKHKE UVTGUU TGURQPUG RTQVGKPU. %TWEKCN EGNN GPXGNQRG RTQVGKPU KPXQNXGF KP CURGEVU QH UVTGUU TGURQPUG CPF UWTXKXCN CTG KPFKECVGF KP VhG ECTVQQP CPF VhG NKUVGF DGNQY þUGG VhG VGZV QH VhG EhCRVGT HQT HWTVhGT FGVCKNU QP KPFKXKFWCN RTQEGUUGU CPF RTQVGKPU).
6CDNG 1. /CETQOQNGEWNGU KP VJG %GNN ©PXGNQRG QH Saccharomyces cerevisiae. %QOÿ RQWPFU EQOOQP VQ VhG EGNN GPXGNQRG QH Saccharomyces cerevisiae þCFCRVGF HTQO XCP FGT 4GUV GV CN. 1ýýõ CPF -NKU GV CN. ú00ú). 6hG KPFKXKFWCN COQWPV QH GCEh EQORQWPF XCTKGU FGÿ RGPFKPI QP VhG Rh[UKQNQIKECN UVCVG QH VhG EGNNU. (QT KPUVCPEG, C EGNN YCNN UVTGUU NGCFU VQ C 10ÿ HQNF KPETGCUG KP EhKVKP EQPVGPV CU OGCUWTGF QP FT[ÿYGKIhV þG.I. 8CNFKXKGUQ GV CN. ú000). # UVTGUU YKVh YGCMÿQTICPKE CEKFU NGCFU VQ C CV NGCUV úÿù HQNF KPFWEVKQP QH VhG KPVGITCN OGOÿ DTCPG RNGKQVTQRKE FTWI TGUKUVCPEG RWOR þ2FT1úR) þ2KRGT GV CN. 1ýýô). /QNGEWNGU EhCTCEVGTKUÿ VKE CPF GUUGPVKCN HQT S. cerevisiae CTG IKXGP KP KVCNKEU. 2NCUOC OGODTCPG ©PGTI[ EQPUGTXCVKQP RTQVGKPU 2NGKQVTQRKE FTWI TGUKUVCPEG RTQVGKPU 2TKOCT[ VTCPURQTV RTQVGKPU %GNN YCNN U[PVhGUK\KPI GP\[OGU ©TIQUVGTQN 2hQURhQNKRKFU 5RhKPIQNKRKFU
%GNN YCNN 5VTWEVWTCN /CPPQRTQVGKPU 5VTGUU /CPPQRTQVGKPU òNKIQUCEEhCTKFG ETQUUÿNKPMKPI GP\[OGU5VTWEVWTCN OGODTCPG RTQVGKPUDGVCÿ1,ñ INWECP DGVCÿ1,3 INWECP EhKVKP
ý ;GCUVU CPF HQQF URQKNCIG
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9.3.1 %GNN YCNN UWICTU CPF VJGKT U[PVJGUKU 6hG EGNN YCNN QH [GCUV EQPUKUVU HQT VhG OCLQT RCTV QH C DGVCÿ1,3ÿNKPMGF INWECP PGVÿ YQTM. ðPFGT TGIWNCT ITQYVh EQPFKVKQPU, VhKU COQWPVU VQ õ0ÿõõó QH VhG EGNN YCNN FT[ÿ YGKIhV þ/CPPGTU GV CN. 1ý73û (NGGV 1ýý1). /GODTCPG NQECNKUGF GP\[OG U[UVGOU EQPVCKPKPI GKVhGT (MU1R þINWECP U[PVhCUG 1) QT üUEúRö(MUúR þINWECP U[PVhCUG ú) FTKXG VhG RTKOCT[ U[PVhGUKU QH VhKU EQORQWPF. 6hKU UOCNN RTQVGKP HCOKN[ KU GUUGPÿ VKCN HQT EQTTGEV DGVCÿ1,3 INWECP DKQU[PVhGUKU þ%CDKD GV CN. 1ýýô). 9hGVhGT VhGUG RTQVGKPU TGRTGUGPV VhG CEVWCN ECVCN[VKE UWDWPKV QH VhG INWECP U[PVhCUG EQORNGZ TGÿ OCKPU VQ DG GUVCDNKUhGF. +V KU HQT GZCORNG TGOCTMCDNG VhCV VhG[ FQ PQV EQPVCKP VhG EQPUGPUWU UGSWGPEG -ö4:üü CU HQWPF KP IN[EQIGP U[PVhCUG CPF 5EhK\QUCEÿ EhCTQO[EGU RQODG CNRhCÿ1,3 INWECP U[PVhCUG þ(WTWMCYC GV CN. 1ýý3û *QEhUVGPÿ DCEh GV CN. 1ýýô). 5WEh OGODTCPG NQECNKUGF RTQVGKPU TGRTGUGPV, IKXGP VhGKT GUUGPÿ VKCN TQNG KP EGNN YCNN DKQIGPGUKU, CP KFGCN VCTIGV HQT VhG FGXGNQROGPV QH CP CPVKHWPICN UVTCVGI[. +PFGGF VhG QTKIKPCN FKUEQXGT[ QH VhG CNVGTPCVKXG INWECP U[Pÿ VhCUG (MUúR YCU HCEKNKVCVGF VhTQWIh VhG UETGGPKPI HQT h[RGTUGPUKVKXKV[ QH fks1 PWNN OWVCPVU HQT VhG CPVKHWPICN GEhKPQECPFKP øÿ733,õñ0 þ/C\WT GV CN. 1ýýõ). /GCPÿ YhKNG, VhG EQPUVKVWVKXG INWECP U[PVhCUG (MU1R hCU DGGP UVWFKGF UWDUVCPVKCNN[ KP C OGFKECN EQPVGZV CU CPVKHWPICN VCTIGV KP 5CEEhCTQO[EGU EGTGXKUKCG, Candida albicans CPF Cryptococcus neoformans þ&QWINCU GV CN. 1ýý7û 6hQORUQP GV CN. 1ýýýû òPKUhK GV CN. ú000û 8KEGPVG GV CN. ú001û 2QRQNQ GV CN. ú001). +PVGTGUVKPIN[, QPG QH VhG EQORQWPFU KFGPVKHKGF KP VhGUG UVWFKGU, GTIQMQPKP #, KU RTQFWEGF D[ C Trichderma URR. C HWPICN IGPWU VQ YhKEh CNUQ VhG KP CITKEWNVWTG WDKSWKVQWUN[ WUGF DKQÿ EQPVTQN HWPIWU Trichoderma harzianum DGNQPIU. 6hG DGVCÿ1,3 INWECP PGVYQTM KU UWDUGSWGPVN[ DTCPEhGF RTGUWOCDN[ VhTQWIh VhG CEVKQP QH GZVTCEGNNWNCT þOGODTCPG CUUQEKCVGF) GPFQVTCPUIN[EQU[NCUGU UWEh CU $INúR þüQNFOCP GV CN. 1ýýõû %CRGNNCTQ GV CN. 1ýýô) CPF üCU1R þ2QRQNQ GV CN. 1ýýýû /QW[PC GV CN. ú000). 5WEh ETQUUÿNKPMKPI GP\[OGU CTG D[ PQY YGNN TGEQIPK\GF CPÿ VKHWPICN VCTIGVU UKPEG VhGKT KPCEVKXCVKQP CU FGOQPUVTCVGF HQT KPUVCPEG KP VhG gas1 PWNN OWVCPV NGCFU VQ UGXGTG RGTVWTDCVKQP QH EGNNWNCT hQOGQUVCUKU þFKUEWUUGF G.I. KP 2QRQNQ CPF 8CK 1ýýý). $GVCÿ1,ñ INWECP KU CPQVhGT EhCTCEVGTKUVKE CPF GUUGPVKCN INWECP RQN[OGT KP DQVh Saccharomyces cerevisiae CPF Candida albicans þUGG G.I. -NKU GV CN. ú00ú). +VU QEÿ EWTTGPEG KP VhG OCLQT HQQF URQKNCIG [GCUV Zygosaccharomyces bailii hCU PQV [GV DGGP UVWFKGF GZVGPUKXGN[. *QYGXGT, VhG HKPFKPI VhCV Z. bailii KU UWUEGRVKDNG VQ VhG CEVKQP QH C EQODKPCVKQP QH C DGVCÿ1,ñ INWECP RQN[OGT CPF C OGODTCPG CEVKXG CIGPV UKOKNCT VQ S. cerevisiae EQTTQDQTCVGU VhG PQVKQP VhCV VhKU [GCUV CNUQ RTQDCDN[ TGNKGU QP DGVCÿ1,ñ INWECP HQT GUUGPVKCN HGCVWTGU QH KVU EGNN GPXGNQRG þ$QO GV CN. ú001 UGG CNUQ DGNQY). $GVCÿ1,ñ INWECP KU RTGUGPV CU C hKIhN[ DTCPEhGF RQN[OGT VhG U[PVhGUKU QH YhKEh RTGUWOCDN[ KPXQNXGU ©4, üQNIK, CPF RNCUOCOGODTCPG NQECNKUGF RTQVGKP EQORNGZGU þ/QPVKLP GV CN. 1ýýýû 5hChKPKCP GV CN. ú000). # ECVCN[VKE UWDWPKV GPEQFÿ KPI IGPG hCU, hQYGXGT, PQV [GV DGGP KFGPVKHKGF. 6hG INWECP RQN[OGT KU XGT[ KORQTÿ VCPV HQT PQTOCN [GCUV YCNN UVTWEVWTKPI UKPEG OWVCPVU CHHGEVGF KP KVU U[PVhGUKU, G.I. kre9 , knh1 CPF skn7 CTG UGXGTGN[ KORCKTGF KP VhGKT EGNNWNCT UVTWEVWTG CPF XKVCNKV[ þUGG G.I. $TQYP GV CN. 1ýý3û &KLMITCCH GV CN. 1ýýñû øWUUKGT GV CN. 1ýýô). +P UVWFKGU
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5. $TWN GV CN.
TGICTFKPI VhG FGXGNQROGPV QH PGY CPVKHWPICN HQQF RTGUGTXCVKQP U[UVGOU $TWN CPF EQÿYQTMGTU RCVGPVGF VhG WUG QH EQODKPCVKQPU QH YCNN N[VKE GP\[OGU EQPVCKPKPI CV NGCUV CNUQ C DGVCÿ1,ñ INWECPCUG þ$TWN GV CN. 1ýý7). 6hG HKPCN EGNN YCNN RQN[OGT EQOOQP VQ CNN HWPIK KU EhKVKP þDGVCÿ1,ù NKPMGF ÷ÿ CEGV[NINWEQUCOKPG). ÷QTOCNN[ VhG [GCUV YCNN EQPVCKPU QP FT[ÿYGKIhV DCUKU 1ÿúó EhKVKP QTICPKUGF KP NKPGCT EhCKPU þ*QNCP GV CN. 1ýô1 UGG CNUQ -NKU GV CN. ú00ú). *QYGXGT, WRQP CRRNKECVKQP QH XCTKQWU HQTOU QH UVTGUU, PQVKEGCDN[ YCNN RGTVWTDCVKQP GKVhGT KP OWVCPVU QT VhTQWIh VhG CEVKQP QH YCNN N[UKPI EQORQWPFU CFFGF HTQO VhG QWVUKFG VhKU KPETGCUGU VQ >10ó QP FT[ÿYGKIhV DCUKU. 6hG OGODTCPGÿNQECNKUGF EhKVKP U[PVhCUGU TGURQPUKDNG HQT EhKVKP U[PVhGUKU KP NCVÿ GTCN YCNNU CPF VhG DWF UKVG DGNQPI VQ C IGPG HCOKN[ YKVh CHS1 VQ CSH3 CU KVU RTQOKPGPV OGODGTU þ%CDKD GV CN. ú001). +V KU GURGEKCNN[ %hU3R VhCV KU TGURQPUKDNG HQT VhG TGRCKT QH YCNN FCOCIG þ8CNFKXKGUQ GV CN. ú000) . 6hKU PQVKQP KU EQTTQDQTCVGF D[ VhG h[RGTUGPUKVKXKV[ QH chs3 OWVCPVU VQ PKMMQO[EKPÿ< þ2QRQNQ CPF 8CK 1ýýý). +PFGGF EhKVKP U[PVhGUKU hCU DGGP CPF UVKNN KU C RTKOG VCTIGV HQT VhG FGXGNQROGPV QH PGY CPVKHWPICNU þüGQTIQRCRCFCMQW CPF 6MCE\ 1ýýõû *QTUEh GV CN. 1ýý7û -NKU GV CN. ú00ú). +P OCP, KV KU PQY MPQYP VhCV VhG KOOWPQNQIKECN FGHGPEG U[UVGO CNUQ RTQÿ FWEGU EhKVKPCUGU UhQYKPI ENGCTN[ VhCV GXGP QWT QYP FGHGPEG KU DCUGF UKIPKHKECPVN[ QP CP CVVCEM QH VhKU HWPICN YCNN RQN[OGT þ$QQV GV CN. 1ýýõû $QQV GV CN. ú001). (Kÿ PCNN[, KP VhG HQQF OCPWHCEVWTKPI KPFWUVT[, EhKVKPCUGU hCXG QHVGP DGGP RTQRQUGF CU CPVKHWPICN CIGPVU KP RCTVKEWNCT KP VhG CTGC QH CITKEWNVWTG þ9CPKUCMC GV CN. ú001). 9.3.2 /GODTCPG NQECNKUGF VTCPURQTVGTU QH NQY OQNGEWNCT YGKIJV EQORQWPFU ÷GZV VQ VhG RTQVGKPU KPXQNXGF KP OCETQOQNGEWNCT U[PVhGUKU, VhGTG CTG PWOGTQWU OGODTCPG NQECNKUGF NQYÿOQNGEWNCT YGKIhV VTCPURQTVGTU, YhKEh HWNHKN GUUGPVKCN EGNÿ NWNCT HWPEVKQPU. # RTKOG ECVGIQT[ QH RTQVGKPU KU VhG hGZQUG VTCPURQTVGTU. ;GCUV EQPÿ VCKPU ú0 hGZQUG VTCPURQTVGTU þHXT1 VQ HXT17, GAL2, SNF3 CPF RGT2) DGNQPIKPI VQ C UWRGTHCOKN[ QH 3ù UWICT RGTOGCUGU þ$QNGU CPF *QNNGPDGTI 1ýý7û UGG CNUQ %hCRVGTU 3 CPF ù). 6hG XCTKQWU hGZQUG VTCPURQTVGTU CTG KPFWEGF FKHHGTGPVKCNN[ FGÿ RGPFKPI QP INWEQUG CXCKNCDKNKV[. +P QTFGT VQ EQORNGVGN[ CDQNKUh ITQYVh QP INWEQUG UVTCKPU YKVh hxt1 VQ hxt17 PWNN OWVCVKQPU CPF CFFKVKQPCNN[ C FGNGVKQP QH VhTGG OGOÿ DGTU QH VhG OCNVQUG VTCPURQTVGT HCOKN[ hCF VQ DG ETGCVGF þò\ECP CPF JQhPUVQP 1ýýýû 9KGE\QTMG GV CN. 1ýýý). 6hG INWEQUG NGXGNU KP VhG GZVTCEGNNWNCT GPXKTQPOGPV TGIWNCVG VhG GZRTGUUKQP CPF HWPEVKQP QH VhG XCTKQWU hGZQUG WRVCMG RTQVGKPU. HXT1 CPF HXT3 GPEQFKPI NQYÿ CHHKPKV[ VTCPURQTVGTU CTG GZRTGUUGF YhGP EGNNU CTG EWNVWTGF KP INWEQUG TKEh EQPFKÿ VKQPU þPQVG VhCV HXT3 KU CNUQ GZRTGUUGF YhGP INWEQUG KU NGUU CDWPFCPV!). +P EQPÿ VTCUV, HXT2 CPF HXT6 GPEQFKPI hKIh CHHKPKV[ VTCPURQTVGTU, CPF HXT4 GPEQFKPI C NQY VQ OQFGTCVG CHHKPKV[ VTCPURQTVGT, CTG GZRTGUUGF YhGP INWEQUG DGEQOGU NKOKVKPI þUGG G.I. 4GKHGPDGTI GV CN. 1ýý7). HXT5 KU GZRTGUUGF KP EhGOQUVCVU CV NQY INWEQUG CPF NQY ITQYVhÿTCVG þUGG G.I. &KGFGTKEh GV CN. 1ýýý). 4GEGPV GXKFGPEG UWIIGUVU KPÿ XQNXGOGPV QH HXT5 KP VhG TGURQPUG QH EGNNU VQ UGXGTG UVTGUU EQPFKVKQPU, KPENWFKPI hGCV UVTGUU, NGCFKPI VQ C TGFWEGF ITQYVh TCVG þ8GTYCCN GV CN. ú00ú). +PVGTGUVKPIN[,
ý ;GCUVU CPF HQQF URQKNCIG
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/GPUQPKFGU GV CN. þú00ú) hCXG TGEGPVN[ UhQYP VhCV WRQP CRRNKECVKQP QH C EQPVKPWÿ QWU hGCV UVTGUU [GCUV EGNNU UhQY CP KPKVKCN DWTUV KP VhGKT INWEQUG HNWZ, RTGUWOCDN[ EQPVTKDWVKPI UKIPKHKECPVN[ VQ CP KPKVKCN UhCTR RGCM KP VTGhCNQUG CEEWOWNCVKQP WRQP hGCV UVTGUU. +V OC[ DG KPHGTTGF VhCV VhG KPFWEVKQP QH HXT5 WRQP hGCV UVTGUU HCEKNKÿ VCVGU VhG TGFKTGEVKQP QH VhG INWEQUG HNWZ HTQO ECVCDQNKE TQWVGU VQ VhG U[PVhGUKU QH VTGhCNQUG. 9hGVhGT VhKU VCMGU RNCEG TGOCKPU VQ DG UVWFKGF KP FGVCKN. +PVGTGUVKPIN[, CP hxt5 PWNN OWVCPV FKURNC[U TGFWEGF HKVPGUU FWTKPI ITQYVh CV R* ô, ITQYVh KP OKPKOCN OGFKC, CPF VhG CFFKVKQP QH WR VQ 1 / UQFKWO EhNQTKFG CU FGVGTOKPGF D[ NCTIGÿUECNG EQORCTCVKXG DCVEh EWNVWTG CUUC[U þüKCGXGT GV CN. ú00ú). +P XKGY QH VhG ETWEKCN TQNG QH *ZV RNCUOCOGODTCPG RTQVGKPU, VhG[ CTG RTKOG VCTIGVU QH CPVKHWPICN UVWF[. +P RCTVKEWNCT, KP XKGY QH KVU RWVCVKXG MG[ TQNG KP UVTGUU UWTXKXCN *ZVõR EQWNF DG CP KORQTVCPV CPVKHWPICN VCTIGV. 6hKU KU UVKNN TGNCVKXGN[ WPÿ GZRNQTGF KP RCTVKEWNCT KP UVWFKGU YKVh TGHGTGPEG VQ VhG HQQF KPFWUVT[. # OCLQT EQPÿ EGTP KU VhCV VhG GZVTCEGNNWNCT TGIKQPU QH VhG VTCPURQTVGT RTQVGKPU, YhKEh CTG OQUV GCUKN[ CEEGUUKDNG, hCXG C HQNF VhCV KU XGT[ UKOKNCT VQ UVTWEVWTGU QEEWTTKPI KP hKIhGT GWMCT[QVGU þOCP) UWEh VhCV KUUWGU QH ETQUU TGCEVKXKV[ CPF VQZKEKV[ YQWNF KOOGFKÿ CVGN[ DG GXKFGPV. +P CFFKVKQP VQ VhG *ZVU, VhGTG KU C OCLQT HCOKN[ QH RTQVGKPU, EQNNGEVKXGN[ ECNNGF RNGKQVTQRKE FTWI TGUKUVCPEG þ2FT) RTQVGKPU VhCV QEEWT KP VhG [GCUV RNCUOC OGODTCPG CPF RNC[ CP KORQTVCPV TQNG KP UVTGUU UWTXKXCN. 6hKU PDR IGPGÿHCOKN[ KU KPXQNXGF KP GZVTWUKQP QH HQTGKIP EQORQWPFU CU FKXGTUG CU ENKPKECNN[ CPF CITKEWNVWTCNN[ KORQTÿ VCPV HWPIKEKFGU, OCP[ CPVKDKQVKEU, hGTDKEKFGU, CPF GXGP OCLQT HQQF RTGUGTXCVKXGU þDGP\QKE CEKF CPF UQTDKE CEKF) þTGXKGYGF KP -QNCE\MQYUMK GV CN. 1ýýôû 2KRGT GV CN. ú001). GZVTCEGNNWNCT VCTIGV NQQRU
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(KI. ÿ. +PHGTTGF OGODTCPG VQRQNQI[ QH VhG YGCM CEKF UVTGUU TGUKUVCPEG RTQVGKPU 2FT1úR. +Pÿ FKECVGF CTG VhG GZVTCEGNNWNCT NQQRU VhCV OKIhV EQPUVKVWVG URGEKHKE VCTIGV UKVGU HQT CPVKÿ[GCUV CIGPVU. &CVC VCMGP HTQO ;2& WUKPI UVCPFCTF RTQVGKP VQRQNQI[ RTGFKEVKQP UQHVYCTG.
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5. $TWN GV CN.
6hG [GCUV 2FT RTQVGKPU CTG CNN #$%ÿVTCPURQTVGT RTQVGKPU YKVh C EQPUGTXGF #62ÿ DKPFKPI ECUUGVVG. %QORQWPFU KPENWFG XCTKQWU EhGOKECN UVTWEVWTGU UWEh CU R[TKOKFKPGU, DGP\KOKFC\QNGU, KOKFQVhKC\QNGU, DKNG CEKFU, CPF WPUCVWTCVGF HCVV[ CEÿ KFU þKPENWFKPI UQTDKE CEKF). 2FT1úR YCU UhQYP VQ DG KPFWEGF WRQP ITQYVh QH [GCUVU KP VhG RTGUGPEG QH KPETGCUKPI COQWPVU QH UQTDKE CEKF þ2KRGT GV CN. 1ýýô). +P CFFKVKQP, KV YCU UhQYP VhCV C FGNGVKQP QH VhG IGPG NGCFU VQ UKIPKHKECPVN[ KPETGCUGF UQTDKE CEKF UGPUKVKXKV[. 2FT1úR KU WPKSWG VQ [GCUV, FQGU PQV QEEWT KP OCP CPF hGPEG KVU GZVTCEGNNWNCT NQQRU CTG CVVTCEVKXG VCTIGVU HQT UGNGEVKXG DKPFKPI QH CPVKOKETQDKCN CIGPVU þ(KI. ú). 5VWFKGU VQ KFGPVKH[ UWEh EQORQWPFU CTG EWTTGPVN[ DGKPI RGTHQTOGF þ2CRCFKOKVTKQW /, RGTUQPCN EQOOWPKECVKQP). (WTVhGT UVWF[ D[ FG ÷QDGN CPF EQÿ YQTMGTU CPF QVhGTU hCU, PQPGVhGNGUU, UhQYP VhCV VhKU KU ENGCTN[ PQV VhG QPN[ EGNNWNCT RTQEGUU MG[ VQ YGCMÿCEKF TGUKUVCPEG þFG ÷QDGN GV CN. ú001û FKUEWUUGF GZVGPUKXGN[ KP 2KRGT GV CN. ú001). +P CFFKVKQP, CEVKXCVKQP QH EGNN YCNN OCKPVGPCPEG U[UVGOU CPF OGODTCPG NQECNKUGF UVTGUU RTQVGKPU KU QDUGTXGF. (KPCNN[, KV KU GUUGPVKCN VhCV VhG OGODTCPG NQECNKUGF 2ÿV[RG RTQVQP RWORKPI #62ÿCUG KU RTGUGPV CPF CEVKXG KP EGNNU TGURQPFKPI VQ YGCM CEKF UVTGUU. 6hG 2FT1ú RTQVGKP RWORU VhG YGCM CEKF CPKQPU VQ VhG GZVTCEGNNWNCT GPXKTQPOGPV þ*QN[QCM GV CN. 1ýýý) YhKNG VhG RTQVQP RWORKPI #62ÿCUG GZVTWFGU VhG GZEGUU RTQVQPU HTQO VhG E[VQUQN þUGG 2KRGT GV CN. 1ýý7). ©CEh QH VhGUG OGODTCPG NQECNKUGF VTCPURQTV RTQVGKPU RTGUGPVU RTQOKUKPI VCTIGVU HQT VhG FGXGNQROGPV QH PGY CPVKHWPICN CRRTQCEhGU. #ICKP VhQWIh, VhG KUUWG QH UKIPKHKECPV UVTWEVWTCN hQOQNQI[ DGVYGGP VhG [GCUV #62ÿCUG CPF UKOKNCT OQNGEWNGU KP hKIhGT GWMCT[QVGU CPF RNCPVU þUGG G.I. #ODGUK GV CN. 1ýýý) RTGENWFGF GZRNQTCVKQP QH VhKU RTQVGKP CU CPVKHWPICN VCTIGV KP FGVCKN þDQVh HQT HQQF CPF OGFKECN CRRNKECVKQPU). òPN[ TGNCVKXGN[ hCU KV DGGP HQWPF VhCV CNVhQWIh XGT[ hQOQNQIQWU, VhG RTQVGKP KU PQV KFGPÿ VKECN VQ VhG OCOOCNKCP EQWPVGTRCTV. *GPEG, KP RTKPEKRNG VhGTG KU UEQRG HQT VhG FGÿ XGNQROGPV QH PQPÿQXGTVN[ VQZKE KPhKDKVQTU. üKXGP VhG ETWEKCN TQNG VhCV VhG RTQVGKP RNC[U KP [GCUV hQOGQUVCUKU PQV QPN[ WPFGT YGCM CEKF UVTGUU DWV CNUQ WPFGT QVhGT UVTGUUGU KPENWFKPI hGCV UhQEM þUGG G.I. 2CPCTGVQW CPF 2KRGT 1ýýú), KVU GZRNQKVCVKQP CU CPVKHWPICN VCTIGV KPFGGF TGOCKPU CP CVVTCEVKXG QRVKQP. (QT GZCORNG, /CPCXCVhW GV CN. þú001) hCXG UhQYP KP VhG OGFKECNN[ TGNGXCPV [GCUV Cryptococcus neoformans VhCV VhG EQPLWICVGF UV[T[N MGVQPG ÷%117õ hCU CU QPG QH KVU EGNNWNCT VCTIGVU VhG RNCUOC OGODTCPG RTQVQP RWORKPI #62ÿCUG. 9.3.3 %GNN YCNN RTQVGKPU CPF UVTGUU TGURQPUG 6hG EGNN GPXGNQRG EQPVCKPU XCTKQWU EGNN YCNN RTQVGKPU KORQTVCPV HQT KVU UVTWEVWTG. 5QOG QH VhGUG CTG CNUQ KPFWEGF UKIPKHKECPVN[ WRQP UVTGUU TGURQPUG þFKUEWUUGF GZÿ VGPUKXGN[ KP -NKU GV CN. ú00ú). 6hG EGNN YCNN EQPVCKPU VYQ ENCUUGU QH RTQVGKPU, VhG IN[EQU[NRhQURhCVKF[NKPQUKVQN þü2+)ÿCPEhQTGF EGNN YCNN RTQVGKPU CPF VhG RTQVGKPU YKVh KPVGTPCN TGRGCVU þ2+4) CPEhQTGF VQ VhG YCNN DGVCÿ1,3 INWECP KP CP CU [GV WPÿ EhCTCEVGTKUGF OCPPGT þ-CRVG[P GV CN. 1ýýý). 6hG NCVVGT RTQVGKPU CTG CNN UVTWEVWTGF CEEQTFKPI VQ VhG HQNNQYKPI UGSWGPEG 52ÿ-GZúÿTGRGCVþU)ÿ%:þññ)%:þ1ñ)%:þ1ú)% KPFKECVKPI C UKIPCN RGRVKFG HQNNQYGF D[ C -GZú UKVG HQNNQYGF D[ XCTKQWU TGRGCVU. (QT C FGVCKNGF FKUEWUUKQP QP VhG UVTWEVWTG QH VhG ü2+ CPF 2+4 RTQVGKPU, VhGKT EQPÿ VTKDWVKQP VQ YCNN RQTQUKVK[ CPF VhG IGPGU GPEQFKPI VhGUG RTQVGKPU VhG TGCFGT KU TGÿ
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HGTTGF VQ þ6QhÿG GV CN. 1ýý3û %CTQ GV CN. 1ýý7û /TUC GV CN. 1ýýýû -CRVG[P GV CN. 1ýýý). +P QTFGT VQ IGV C HWNN RKEVWTG QH VhG [GCUV EGNN YCNN RTQVGKPU, VhG CKF QH CNIQÿ TKVhOU RTGFKEVKPI VhGKT EGNNWNCT NQECNKUCVKQP DCUGF QP RTQVGKP UVTWEVWTG KU DGEQOKPI KPETGCUKPIN[ RQRWNCT. 4GEGPVN[ FG üTQQV GV CN. þú003) hCXG FGHKPGF CP CNIQTKVhO VhCV KFGPVKHKGF ñý RWVCVKXG ü2+ÿCPEhQTGF EGNN YCNN RTQVGKPU KP Saccharomyces cerevisiae . 5WEh RTGFKEVKQPU PQY CYCKV GZRGTKOGPVCN XGTKHKECVKQP. %GNN YCNN RTQVGKPU RNC[ CP KORQTVCPV TQNG KP FGNKOKVKPI VhG RQTQUKV[ QH VhG YCNN CPF RTQXKFKPI KV YKVh KVU EhCTCEVGTKUVKE UVTWEVWTG CPF EhCTIG. +P GNGEVTQP OKETQUÿ EQR[ VhG[ UVCKP FCTM CPF IKXG VhG EGNN YCNN KVU EhCTCEVGTKUVKE GNGEVTQPÿFGPUG QWVGT TKPI UVTWEVWTG þUGG G.I. /CTSWCTFV 1ýñú). %GNN YCNN RTQVGKPU CTG hGCXKN[ IN[EQU[ÿ NCVGF. 6hG ÷ÿ CPF òÿNKPMGF QNKIQUCEEhCTKFGU OC[ OCMG WR VQ UQOGVKOGU ý0ó QH VhG YGKIhV HTCEVKQP QH VhG RTQVGKPU þòTNGCP 1ýý7). +P RCTVKEWNCT, VhG hGCXKN[ DTCPEhGF ÷ÿNKPMGF EhCKPU hCXG CNTGCF[ UKPEG NQPI DGGP KORNKECVGF KP VhG NKOKVKPI QH YCNN RQTQUKV[ þ&G ÷QDGN 1ýý0). %GNN YCNN RTQVGKPU DGNQPIKPI VQ VhG ü2+ÿNKPMGF HCOKN[ YGTG UhQYP VQ DG KPXQNXGF KP UVTGUU RTQVGEVKQP KP UGXGTCN UVWFKGU. (QT KPÿ UVCPEG %YR1R, CPF %YRúR, CTG KPXQNXGF KP VhG RTQVGEVKQP QH EGNNU CICKPUV GP\[ÿ OCVKE CVVCEM CPF CPVKOKETQDKCN RGRVKFGU þXCP FGT 8CCTV 1ýýõû &KGNDCPFhQGUKPI 1ýýô). 6KR1R, 6KT1R, 6KTúR, 6KTùR CTG KPXQNXGF KP VGORGTCVWTG UVTGUU þ&QP\GCW GV CN. 1ýýñ). 6hG EGNN YCNN RTQVGKPU DGNQPIKPI VQ VhG 2+4 HCOKN[ OC[ hCXG CV NGCUV UQOG UKOKNCT TQNGU. +PVGTGUVKPIN[, KV hCU DGGP UhQYP VhCV QXGTGZRTGUUKQP QH VhG 2+4 IGPGU KPFWEGF TGUKUVCPEG VQ VhG RNCPV CPVKHWPICN RTQVGKP QUOQVKP þ24ÿõ CPVKHWPICN RTQVGKP HCOKN[). &GNGVKQP QH CNN PIR IGPGU KP [GCUV NGF VQ C h[RGTUGPUKVKXKV[ QH VhG [GCUV EGNNU VQ QUOQVKP þ;WP GV CN. 1ýý7). 2CTVKEWNCTN[ KPVGTGUVKPI KU VhCV WPFGT EGTVCKP UVTGUU EKTEWOUVCPEGU VhG NKPMCIG QH VhG EGNN YCNN RTQVGKPU VQ VhG YCNN ECP UKIPKHKECPVN[ EhCPIG. 6hG GZVGPV QH VhG DGVCÿ 1,ñ INWECP EhKVKP NKPMCIG KPETGCUGU FTCOCVKECNN[ KP RCTCNNGN YKVh KPETGCUGF EhKVKP EQPVGPV KP EGNN YCNNU QH EGNNU UVTGUUGF YKVh YCNN RGTVWTDKPI EQPFKVKQPU þFKUEWUUGF GZVGPUKXGN[ KP -NKU GV CN. ú00ú). %YR1R GXGP WPFGT PQTOCN EKTEWOUVCPEGU OC[ hCXG C FQWDNG YCNN NKPMCIG VhTQWIh VhG DGVCÿ1,ñ INWECP ü2+ NKPMCIG CPF VhTQWIh VhG 2+4ÿNKMG CNMCNK UGPUKVKXG FKTGEV NKPMCIG VQ DGVCÿ1,3 INWECP þ-CRVG[P GV CN. ú001). 6hG TGURQPUKDNG ETQUUÿNKPMKPI GP\[OGU TGOCKP VQ DG KFGPVKHKGF. +PVGTGUVKPIN[ CNUQ, $QO GV CN. þú001) hCXG UhQYP VhCV VTGCVOGPV QH EGNNU YKVh VhG DGVCÿ1,ñ FKUCEEhCTKFG IGPVKQDKQUG NGCFU VQ C UKIPKHKECPVN[ KPETGCUGF UGETGVKQP QH VhG ü2+ FGRGPFGPV TGRQTVGT RTQVGKP CNRhCÿICNCEVQUKFCUGÿ%YRúR, CPF VQ C UKIPKHKÿ ECPVN[ GPhCPEGF <[OQN[CUG UGPUKVKXKV[. <[OQN[CUG EQPUKUVU QH C EQEMVCKN QH DGVCÿ 1,3 INWECPCUG CPF RTQVGCUG. 6hG CEEGUUKDKNKV[ QH VhG INWECPCUG VQ VhG EGNN YCNN DGVCÿ 1,3 INWECP KU FGVGTOKPGF D[ VhG CDQXG FKUEWUUGF hGCXKN[ IN[EQU[NCVGF OCPPQRTQÿ VGKPU þ&G ÷QDGN GV CN. 1ýý0û òXCNNG GV CN. 1ýýô). 6hWU KV OC[ DG KPHGTTGF VhCV CFFKÿ VKQP QH VhG DGVCÿ1,ñ INWECP KPVGTHGTGF YKVh VhG PQTOCN HWPEVKQP QH C ETQUUÿNKPMKPI GP\[OG CPF hGPEG NGF VQ VhG UGETGVKQP QH VhG EGNN YCNN TGRQTVGTÿRTQVGKP. 6hG GOGTIKPI QXGTCNN RKEVWTG UhQYU VhCV VhG ETQUUÿNKPMKPI QH CNN QH VhG EGNN YCNN RTQVGKPU VQ VhG EGNN YCNN RQN[UCEEhCTKFGU QEEWTU GZVTCEGNNWNCTN[. 6hG ETQUUÿNKPMKPI GP\[OGU CTG MG[ KP VhKU RTQEGUU CPF VhWU CTG KORQTVCPV RTKOG VCTIGVU HQT VhG FGXGNÿ QROGPV QH PGY CPVKHWPICN UVTCVGIKGU UWKVGF DQVh HQT OGFKECN CPF HQT HQQF RTGUGTXCÿ VKQP RWTRQUGU.
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5. $TWN GV CN.
9.3.4 5GPUKPI VJG GZVTCEGNNWNCT GPXKTQPOGPV CPF UKIPCNNKPI UVTGUU. +PVGITCN OGODTCPG RTQVGKPU CTG IGPGTCNN[ RTGUWOGF VQ DG VhG UGPUQTU QH VhG RGTÿ VWTDCVKQPU KP EGNN YCNN CPF OGODTCPG ECWUGF D[ GPXKTQPOGPVCN UVTGUU. +H UWEh RTQÿ VGKPU ECP DG VCTIGVGF KP CPVKHWPICN CRRTQCEhGU, VhG GHHGEVU OC[ DG FWG VQ VhG GUUGPÿ VKCN TQNG QH VhGUG RTQVGKPU KP VhG QPUGV QH VhG UVTGUU TGURQPUG. 5VTGUU TGURQPUG UKIPCN VTCPUFWEVKQP RCVhYC[U KP [GCUV CTG IGPGTCNN[ QH VhG /KVQIGP #EVKXCVGF 2TQVGKP þ/#2) MKPCUG V[RG þFKUEWUUGF GZVGPUKXGN[ KP *QhOCPP ú00ú CPF -NKU GV CN. ú00ú). 6hG RCVhYC[ FKTGEVN[ KPXQNXGF KP EGNN FCOCIG KU VhG 5NVúö/#2 MKPCUG TQWVG, PCOGF CHVGT KVU RTKOCT[ QWVRWV, VhG RhQURhQT[NCVGF /#2 MKPCUG 5NVú þUGG G.I. #Pÿ FTGYU ú000). %WTTGPVN[ MPQYP OGODTCPG NQECNKUGF RTQVGKPU KPXQNXGF KP VhG CEVKÿ XCVKQP QH VhG 5NVú UKIPCNNKPI RCVhYC[ DGNQPI VQ VYQ HCOKNKGUû VhG 95% IGPGÿ HCOKN[ GPEQFKPI 9UE1Rö5NI1R, 9UEúRö3RöùR CPF VhG MID2 IGPGÿHCOKN[ GPEQFKPI /KFúR CPF /VN1R þ8GTPC GV CN. 1ýý7û 4CLCXGN, 1ýýýû -GVGNC GV CN. 1ýýý) . 9hGP VhG EGNN YCNN INWECPÿEhKVKP OGUhYQTM KU CVVCEMGF D[ YCNN N[VKE GP\[OGU, /KFúR OGFKÿ CVGU UKIPCNNKPI QH VhG TGUWNVKPI UVTGUU CPF VhG QPUGV QH 5NVúR CEVKXCVKQP þUGG G.I. &G ÷QDGN GV CN. ú000). 5VTGUU UWTXKXCN QH EGNNU KPEWDCVGF KP VhG RTGUGPEG QH h[FTQIGP RGTQZKFG CPF GVhCPQN KPXQNXGU WSC1/SLG1 CPF WCS 2/3/4 þ<W GV CN. ú001). ©VhCÿ PQN KU IGPGTCNN[ UGGP CU C OGODTCPG RGTVWTDKPI CIGPV þG.I. /CT\C GV CN. ú00ú). +V KU CNUQ IGPGTCNN[ CEEGRVGF VhCV C EGNN YCNN YGCMGPKPI NGCFU VQ OGODTCPG RGTVWTDCVKQP CU C EQPUGSWGPEG QH VhG KPVGTPCN VWTIQT RTGUUWTG. +PVGTGUVKPIN[ VhG YGCMÿQTICPKE CEKF UQTDKE CEKF hCU FWG VQ KVU NKRQRhKNKE PCVWTG CNUQ C hKIh CHHKPKV[ HQT OGODTCPGU. 5VTCVHQTF CPF #PUNQY CNTGCF[ KP 1ýýô RTGUGPVGF GXKFGPEG VhCV UQTDKE CEKF hCU C FWCN OQFG QH CEVKQP K.G. CU C OGODTCPG RGTVWTDKPI CIGPV CPF CU C YGCM QTICPKE CEKF þ5VTCVHQTF CPF #PUNQY, 1ýýô). 9G PQY UGG VhCV C RGTVWTDGF EGNNWNCT hQOGQÿ UVCUKU D[ UQTDKE CEKF KP HCEV NGCFU VQ C UGTKGU QH EhCPIGU CPF CFCRVCVKQPU. &G ÷QDGN GV CN þú001) hCXG UhQYP VhCV KP C IGPQOGÿYKFG UETGGP HQT UQTDKE CEKF KPFWEGF IGPGU KP TGUKUVCPV EGNNU, IGPGU CEVKXG KP VhG 5NVúR RCVhYC[ YGTG ENGCTN[ WRTGIWÿ NCVGF. 9hKEh OGODTCPG UVTGUU UGPUQTU CTG KPXQNXGF KP UGPUKPI UQTDKE CEKF KPFWEGF OGODTCPG RGTVWTDCVKQP TGOCKPU VQ DG KFGPVKHKGF. +P CFFKVKQP, UVWFKGU CTG WPFGTYC[ VQ CUUGUU YhGVhGT VhKU U[UVGO YQTMU KP [GCUV KP IGPGTCN, K.G. KP VhG EQOOQP HQQF URQKNCIG [GCUV Zygosaccharomyces bailii. 6hKU GZCORNG QH VhG WUG QH OQFGTP IGPQOGÿYKFG VTCPUETKRV CPCN[UKU KU QPG QH VhG HKTUV QH KVU MKPF KP VhG CTGC QH HQQF OKETQDKQNQI[ þUGG %hCRVGT 11). +V ENGCTN[ UhQYU VhG ITGCV RQVGPVKCN VhCV IGPQOKEU CPCN[UGU QHHGT KP QTFGT VQ QDVCKP C hQNKUVKE XKGY QP EGNNWNCT RTQEGUUGU KPENWFKPI VhQUG KPXQNXGF KP HQQF RTGUGTXCVKQP. #FGÿ SWCVG $KQ+PHQTOCVKEU CPF FCVCÿCPCN[UKU KU RCTCOQWPV VQ IGVVKPI VhG OCZKOWO XCNWG QWV QH VhGUG GZRGTKOGPVU. 6hG PGZV RCTCITCRh hKIhNKIhVU EWTTGPV FGXGNQRÿ OGPVU KP VhCV HKGNF. 6hG VQQNU FKUEWUUGF CTG IGPGTCNN[ CRRNKECDNG KP IGPQOKEU CPCN[UKU CPF CTG hGTG GZGORNKHKGF YKVh GZCORNGU HTQO UVTGUU TGUKUVCPEG TGUGCTEh KP HQQF RTGUGTXCVKQP.
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9.4. # PGY OGVJQF HQT VJG CPCN[UKU QH IGPG GZRTGUUKQP RTQHKNGU 2GThCRU PQV UWTRTKUKPIN[, Saccharomyces cerevisiae hCU DGEQOG C OQFGN QTICPKUO HQT FGXGNQRKPI DKQKPHQTOCVKEU VQQNU. +V KU VhG HKTUV GWMCT[QVKE QTICPKUO VhCV hCU DGGP HWNN[ UGSWGPEGF. +VU IGPGU hCXG DGGP GZVGPUKXGN[ CPPQVCVGF, CPF KV KU CNUQ VhG HKTUV QTICPKUO HQT YhKEh EQORNGVG IGPG GZRTGUUKQP RTQHKNGU hCXG DGGP QDVCKPGF þ&G4KUK GV CN. 1ýý7). +P CFFKVKQP, KV KU RQUUKDNG VQ UVWF[ VhG HKVPGUU QH C HWNN UGV QH FGNGVKQP OWVCPVU KP C UKPING GZRGTKOGPV D[ WUKPI ÂDCTÿEQFGF• UVTCKPU þüKCGXGT GV CN. ú00ú). 6hG PWODGT QH VTCPUETKRV RTQHKNKPI GZRGTKOGPVU KU TCRKFN[ ITQYKPI CPF CV RTGUGPV CNTGCF[ GZEGGFU UGXGTCN VhQWUCPFU. 5GXGTCN CRRTQCEhGU hCXG DGGP FGXGNÿ QRGF VQ CPCN[\G VhG FCVC QDVCKPGF. +P RCTVKEWNCT, VhG hKGTCTEhKECN ENWUVGT CPCN[UKU hCU DGEQOG RQRWNCT þ©KUGP GV CN. 1ýýô). #NVhQWIh KV KU C RQYGTHWN OGVhQF, KVU FKUÿ CFXCPVCIGU CTG VhCV OWNVKRNG GZRGTKOGPVU CTG PGGFGF CPF VhG WUG QH CTDKVTCT[ EWVÿ QHHU HQT FGHKPKPI WRÿ CPF FQYPÿTGIWNCVGF IGPGU. *GTG YG FKUEWUU C PGY OGVhQF, 4©&ð%©, YhKEh KU CDNG VQ CPCN[\G UKPING IGPG GZRTGUUKQP RTQHKNGU CPF CNUQ CNNQYU FKTGEV KFGPVKHKECVKQP QH RWVCVKXG WRUVTGCO TGIWNCVQT[ UGSWGPEGU CPF SWCPVKHKECVKQP QH VhGKT EQPVTKDWVKQP VQ IGPG GZRTGUUKQP NGXGNU. 4©&ð%© KU UhQTV HQT 4GIWNCVQT[ ©NGOGPV &GVGEVKQP ðUKPI %QTTGNCVKQP YKVh ©ZRTGUUKQP CPF YCU FGXGNQRGF D[ $WUUGOCMGT CPF EQÿYQTMGTU þ$WUUGOCMGT GV CN. ú001, YYY.EQNWODKC.GFWöEWöDKQNQI[öHCEWNV[öDWUUGOCMGTö). 6hKU OGVhQF EQTTGÿ NCVGU VhG RTGUGPEG QH RCTVKEWNCT WRUVTGCO OQVKHU YKVh VhG QDUGTXGF EhCPIGU KP IGPG GZRTGUUKQP. +V KU DCUGF QP C UKORNG NKPGCT OQFGN VhCV RTGFKEVU VhG GZRTGUUKQP NGXGN CUUQEKCVGF YKVh VhG WRUVTGCO UGSWGPEG QH C IKXGP IGPG DCUGF QP VhG TGIWNCVQT[ GNGOGPVU KP VhG UGSWGPEG. # HKVVKPI RCTCOGVGT ( SWCPVKHKGU VhG EQPVTKDWVKQP QH GCEh &÷# OQVKH VQ VhG GZRTGUUKQP NGXGN. (QT GCEh OQVKH, VhG UKIPKHKECPEG QH ^(^ KU SWCPÿ VKHKGF D[ C 2ÿXCNWG, TGRTGUGPVKPI VhG RTQDCDKNKV[ VhCV VhKU XCNWG QH ^(^ QEEWTU D[ EhCPEG. +V RTQXKFGU C WUGHWN CNVGTPCVKXG VQ EQPXGPVKQPCN OQVKH HKPFKPI CRRTQCEhGU, YhKEh YQTM D[ HKTUV ENWUVGTKPI IGPGU DCUGF QP VhGKT GZRTGUUKQP RTQHKNG CETQUU OWNÿ VKRNG GZRGTKOGPVU CPF UWDUGSWGPVN[ HKPFKPI QXGTÿTGRTGUGPVGF OQVKHU KP VhG WRÿ UVTGCO TGIKQPU HQT GCEh ENWUVGT. 6Q KNNWUVTCVG VhG RQYGT QH 4©&ð%©, YG hCXG TGÿCPCN[\GF VhG FCVC D[ 5ChCTC GV CN. þú00ú), YhQ hCXG OQPKVQTGF KP VKOG VhG VTCPUETKRVKQPCN TGURQPUG QH GZRQPGPÿ VKCNN[ ITQYKPI [GCUV EGNNU VTCPUHGTTGF HTQO 30¸% VQ 10¸% þ(KI. 3). ðUKPI 4©&ð%©, YG hCXG KFGPVKHKGF CP CXGTCIG QH VGP FKHHGTGPV CPF UKIPKHKECPV TGIWNCVQT[ OQVKHU RGT VKOG RQKPV. òPG QH VhG GNGOGPVU KU VhG TGXGTUGF EQORNGOGPV þü#ü66ü6) QH VhG øQY òZ[IGP 4GURQPUG ©NGOGPV þøò4©) þ8CUEQPEGNNGU GV CN. ú001). 6hG RWVCVKXG NQY VGORGTCVWTG CPF QZ[IGP UGPUQT /IC1R þ÷CMCICYC GV CN. ú00ú) DKPFU VQ VhG øò4© OQVKH CPF KVU VTCPUETKRV KU KPFWEGF KP VhG EQNF UhQEM GZRGTKOGPV CHVGT 30 OKPWVGU CPF TGOCKPU KPFWEGF FWTKPI VhG TGUV QH VhG VKOG EQWTUG. +PVGTGUVKPIN[, QPG QH VhG IGPGU WPFGT EQPVTQN QH VhG øò4© KU OLE1, YhKEh GPEQFGU C FGNVCÿýÿHCVV[ CEKF FGUCVWTCUG. 6hKU KU EQPUKUVGPV YKVh VhG QDUGTXCVKQP VhCV CV NQY ITQYVh VGOÿ RGTCVWTGU EGNNU KPETGCUG VhG NGXGN QH WPUCVWTCVGF HCVV[ CEKFU KP VhGKT OGODTCPGU KP CP CVVGORV VQ OCKPVCKP OGODTCPG HNWKFKV[ þ÷CMCICYC GV CN. ú00ú). (KIWTG 3 UhQYU
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5. $TWN GV CN. 0.6 0.5 0.4 0.3 0.2 0.1 0 -0.1 -0.2 -0.3
0
100
200
300
400
500
6KOG (OKPþ
(KI. 3. &[PCOKEU QH VhG VTCPUETKRVKQPCN TGURQPUG QH [GCUV EGNNU VQ C EQNF UhKHV. +FGPVKHKECVKQP QH WRUVTGCO TGIWNCVQT[ GNGOGPVU CPF VhGKT EQPVTKDWVKQP VQ IGPG GZRTGUUKQP. #P GZRQPGPÿ VKCNN[ ITQYKPI [GCUV EWNVWTG YCU VTCPUHGTTGF HTQO 30¸% VQ 10¸%. 5CORNGU YGTG VCMGP HQT VTCPUETKRV CPCN[UKU CHVGT 0.úõ, 0.õ, ú, ù, CPF ô h þ5ChCTC GV CN. ú00ú). 6TCPUETKRV TCVKQU YGTG TGÿCPCN[\GF WUKPI 4©&ð%©. 6hG (ÿXCNWGU QH VhG TGIWNCVQT[ WRUVTGCO GNGOGPVU TGRTGUGPV VhG ECNEWNCVGF EQPVTKDWVKQPU QH VhGUG GNGOGPVU VQ IGPG GZRTGUUKQP. 6hG HQWT TGIWNCVQT[ WRÿ UVTGCO OQVKHU UGNGEVGF HQT VhKU ITCRh CTG %ü#6ü#ü þ'), YhKEh KU KPXQNXGF KP VhG TGIWNCÿ VKQP QH T4÷# U[PVhGUKU, #üüüü QT 564© GNGOGPV þ!), YhKEh KU DQWPF D[ VhG /UPúöù VTCPUETKRVKQP HCEVQTU, %%ü6#%# þ"), VhG DKPFKPI UKVG HQT 4CR1, YhKEh KU COQPI QVhGTU KPÿ XQNXGF KP VTCPUETKRVKQPCN CEVKXCVKQP QH IGPGU GPEQFKPI TKDQUQOCN RTQVGKPU, CPF VhG øQY òZ[IGP 4GURQPUG ©NGOGPV þøò4©), ü#ü66ü6 þ8).
VhG F[PCOKEU QH VhG VTCPUETKRVKQPCN TGURQPUG VQ EQNF UhKHV KP VKOG YKVh TGURGEV VQ HQWT TGIWNCVQT[ OQVKHU. 6hGUG YGTG UGNGEVGF HQT VhGKT EhCTCEVGTKUVKE TGURQPUG. #V NGCUV VhTGG FKHHGTGPV V[RGU QH TGURQPUGU ECP DG FKUVKPIWKUhGFï þK) üGPGU EQPVCKPKPI VhG WRUVTGCO OQVKH %ü#6ü#ü, YhKEh KU KPXQNXGF KP VhG TGIWNCVKQP QH T4÷# U[Pÿ VhGUKU, CTG TCRKFN[ WRTGIWNCVGF, DWV KP C NCVGT UVCIG VhG[ CTG TGRTGUUGF. þKK) 6hG EQPÿ VTKDWVKQP QH VhG 4CR1 GNGOGPV, YhKEh KU COQPI QVhGTU KPXQNXGF KP VTCPUETKRVKQPCN CEVKXCVKQP QH IGPGU GPEQFKPI TKDQUQOCN RTQVGKPU, TGCEhGU KVU OCZKOWO KP C NCVGT UVCIG, DWV VhGP FGENKPGU UVTQPIN[ CU YGNN. þKKK) 6hG EQPVTKDWVKQPU QH VhG 564© þ5VTGUU 4GURQPUG ©NGOGPV) OQVKH CPF QH VhG øQY òZ[IGP 4GURQPUG ©NGOGPV þøò4©) KPETGCUG UNQYN[ KP VKOG CPF CTG UVKNN TGNCVKXGN[ hKIh ô hQWTU CHVGT VhG UhKHV VQ 10¸%. 6hG CFXCPVCIGU QH 4©&ð%© CTG VhTGGÿHQNF. (KTUV, 4©&ð%© PGGFU QPN[ C UKPING IGPG GZRTGUUKQP RTQHKNG HQT CPCN[UKU. 5GEQPF, KV KFGPVKHKGU VhG WRUVTGCO TGIWNCVQT[ GNGOGPVU KPXQNXGF KP VhG VTCPUETKRVKQPCN TGURQPUG. 6hKTF, KV SWCPVKHKGU VhG EQPVTKDWÿ VKQPU QH VhG UGRCTCVG GNGOGPVU VQ VhKU TGURQPUG. +V CNUQ CNNQYU HQT C UKORNG QXGTXKGY QH VhG F[PCOKEU QH IGPG VTCPUETKRVKQP, KP EQODKPCVKQP YKVh VhG RQUUKDKNKV[ VQ FKÿ TGEVN[ KFGPVKH[ IGPGU VhCV EQPVTKDWVG VQ HKVPGUU KP URGEKHKE GPXKTQPOGPVCN EQPFKVKQPU þüKCGXGT GV CN. ú00ú). 6hKU YKNN QRGP VhG YC[ HQT VhG HQQF KPFWUVT[ VQ C OWEh FGGRGT WPFGTUVCPFKPI QH VhG hWTFNG VGEhPQNQI[ CPF C OQTG TCRKF CPF TCVKQPCN FGÿ
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UKIP QH PGY EQODKPCVKQPU QH CPVKOKETQDKCN VTGCVOGPVU. #NVhQWIh VhKU CNIQTKVhO hCU DGGP FGUKIPGF HQT S. cerevisiae, KP RTKPEKRNG, VhGTG KU PQ DCTTKGT VQ GZVGPF VhGUG OGVhQFU VQ QVhGT QTICPKUOU YKVh C HWNN[ UGSWGPEGF IGPQOG.
9.÷ %QPENWFKPI TGOCTMU CPF HWVWTG RTQURGEVU ;GCUVU OC[ DG DGPGHKEKCN VQ HQQF RTQFWEVKQP K.G. KP HQQF HGTOGPVCVKQPU UWEh CU VhG RTQFWEVKQP QH DGGT, YKPG, CPF EGTVCKP HGTOGPVGF UQ[ RTQFWEVU. *QYGXGT, VhG UCOG QTICPKUOU OC[ CNUQ ECWUG ITGCV PWKUCPEG. 5RQKNCIG QH FTGUUKPIU, EGTVCKP DGXGTÿ CIGU, CPF FCKT[ RTQFWEVU KU C YGNNÿMPQYP RhGPQOGPQP. +P CFFKVKQP, [GCUVU OC[ ECWUG UKIPKHKECPV hGCNVh KUUWGU KP KOOWPQEQORTQOKUGF hWOCPU. +P QTFGT VQ KPVGTHGTG YKVh VhG ITQYVh QH VhGUG NQYGT GWMCT[QVGU, VhG UGCTEh KU QWV HQT URGEKHKE CPF GUUGPVKCN EGNNWNCT VCTIGVU. 5KPEG VhG EGNNWNCT UVTWEVWTG QH [GCUVU KU UKOKNCT VQ QWTU VhKU KU PQV C VTKXKCN RWTUWKV. -PQYNGFIG QP VhG UVTWEVWTG QH VhG EGNN YCNN CPF EGNNWNCT OGODTCPG UGGO VQ RTQXKFG ENWGU VQYCTFU VhCV IQCN. 6hG CRRNKECVKQP QH OQFGTP þOQNGEWNCT)ÿDKQNQIKECN VQQNU þUWEh CU &÷#ÿOKETQÿ CTTC[U) CNNQYU WU VQ UVWF[ VhG DGhCXKQWT QH OKETQQTICPKUOU CV CP WPRTGEGFGPVGF NGXGN QH FGVCKN CPF UQRhKUVKECVKQP. 6hG GZCORNGU QH VhG WUG QH DKQKPHQTOCVKEU KP CPCN[UKPI IGPGÿGZRTGUUKQP FCVC QDVCKPGF WUKPI OKETQÿCTTC[U KNNWUVTCVG VhG TCRKF FGXGNQROGPVU KP VhG HKGNF. /KETQQTICPKUOU ECP DG XKGYGF CU KPVGITCVGF DKQNQIKECN U[UVGOU VhCV CKO CV UWUÿ VCKPKPI VhGOUGNXGU VhTQWIh GPGTI[ IGPGTCVKQP CPF OCKPVGPCPEG QH GUUGPVKCN UVTWEÿ VWTCN RCTCOGVGTU. 6hG UWTRNWU GPGTI[ IGPGTCVGF KU VhGP WUGF HQT EGNN OWNVKRNKECVKQP, EGNN FKXKUKQP, CPF TGURQPUGU VQ EhCPIKPI GPXKTQPOGPVU. 6hKU EQPEGRV KU PQV FKHHGTÿ GPV HTQO CP[ U[UVGO FGUETKRVKQP CV VhG XCTKQWU NGXGNU QH $KQNQI[. 6hKU KOOGFKCVGN[ DTKPIU HQTYCTF VhG HCEV VhCV KP QTFGT VQ hCXG VhKU KPVGITCN OQNGEWNCT EGNN DKQNQIKECN FGUETKRVKQP QH VhG EGNN, VhGTG KU C PGGF VQ NKPM VhG OQNGEWNCT FGVCKN VQ VhG Rh[UKQÿ NQIKECN FGVCKN þHWPEVKQPCN IGPQOKEUû UGG %hCRVGT 11). 5WEh UVWFKGU CTG EWTTGPVN[ WPFGTVCMGP HQT G.I. VhG TGURQPUG QH S. cerevisiae VQ C EQPVKPWQWU hGCV UVTGUU þ/GPÿ UQPKFGU GV CN. ú00ú). (KTUV TGUWNVU UhQY VhCV WRQP WRÿUhKHV QH VhG EWNVWTG VGORGTCÿ VWTG, EGNNU EWNVWTGF QP hKIh INWEQUG EQPEGPVTCVKQPU VGPF VQ UhQY C TCRKF TKUG KP UWDUVTCVG EQPUWORVKQP HQNNQYGF D[ C FTQR VQ LWUV CDQXG RTGÿUVTGUU NGXGNU, QH VhG UVGCF[ UVCVG UWDUVTCVG EQPUWORVKQP TCVG WRQP TGUVCTV QH ITQYVh QH VhG EGNNU. #NUQ C UVGCF[ KPETGCUG KP DQVh #62 CPF #&2 CDUQNWVG EGNNWNCT NGXGNU YCU PQVGF. (KPCNN[, VTGhCNQUG NGXGNU TQUG UhCTRN[ KOOGFKCVGN[ CHVGT CRRN[KPI VhG VhGTOCN UVTGUU. 6hGUG FCVC RQKPV VQ CP KOOGFKCVG OGVCDQNKE TGURQPUG VQ hGCV UhQEM D[ VhG EGNNU. #HVGT C FGNC[ RhCUG QH CDQWV 0.õÿ1 hQWT VhG EGNN YCNN TGOQFGNNKPI 2ME1R RCVhYC[ YCU VTCPUKGPVN[ CEVKXCVGF CU GXKFGPEGF D[ VhG FWCN RhQURhQT[NCVKQP QH KVU /#2ÿMKPCUG QWVRWV 5NVúR þ/GPUQPKFGU, RGTUQPCN EQOOWPKECVKQP). +PVGTGUVKPIN[, VhG GPF QH VhG CEVKXCVKQP QH VhG RCVhYC[ EQKPEKFGF YKVh VhG GPF QH VhG CEVWCN ITQYVh NCIÿRhCUG. (TQO VhKU, YG KPHGT VhCV CEVKXCVKQP QH VhG RCVhYC[ OKIhV QPN[ DG PGGFGF FWTKPI UVTGUU CFCRVCVKQP VQ CNNQY VhG EGNN VQ TGUVQTG hQOGQUVCUKU CPF VQ KPFWEG CP CFCRVCÿ VKQP OGEhCPKUO. 6hKU UhQWNF HKTUV QH CNN CNNQY VhG EGNN VQ TGCEh C PGY DCNCPEG DGÿ VYGGP KPVTCEGNNWNCT VTGhCNQUG NGXGNU K.G. VWTIQT CPF VhG EGNN YCNN OGFKCVGF EQWPVGT
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5. $TWN GV CN.
HQTEG. 8GTKHKECVKQP QH VhKU h[RQVhGUKU CPF HWTVhGT KORNKECVKQPU CTG EWTTGPVN[ UWDLGEV VQ HWTVhGT UVWF[. +PVGITCVKQP QH OQNGEWNCT CPF OGVCDQNKE FCVC YKNN CNNQY WU VQ DWKNF WR C ÂEQPVKPWWO HWPEVKQP• QH EGNNWNCT DGhCXKQWT DCUGF QP INQDCN EGNNWNCT FCVC HQT IGPG RTGUGPEG, VTCPUETKRVKQP, RTQVGKP RTGUGPEG, HWPEVKQP CPF OGVCDQNKE TGCEVKQPU òPG OCLQT TGOCKPKPI KUUWG KU YhKEh EGNNWNCT HCEVQTU EQPUVKVWVG VhG UYKVEhGU DGÿ VYGGP EGNNÿFKXKUKQP CPF UVTGUU TGURQPUG CPF YhCV FQ VhG[ UGPUG? *QY FQ VhGUG UYKVEhGU KPVGITCVG CNN KPRWV KPHQTOCVKQP, FQ VhG[ QRGTCVG KP C FKIKVCN HCUhKQP QT CPCÿ NQIWG OQFG? +P CFFKVKQP, CNN NKXKPI EGNNWNCT U[UVGOU CTG hGCXKN[ EQORCTVOGPVCNKUGF OQTG QT NGUU. ÷QY VhCV YG ECP FKUUGEV VhG EGNNWNCT Rh[UKQNQI[ CV VhG OQNGEWNCT NGXGN, YG UhQWNF UVCTV CUMKPI YhGTG KP VhG EGNN CNN QH VhGUG TGCEVKQPU QEEWT. # RTQRGT URCVKQÿVGORQTCN WPFGTUVCPFKPI QH UVTGUU TGURQPUG QH OKETQQTICPKUOU WUKPI IGPQOKEU CPF UVCVG QH VhG CTV OKETQUEQRKECN VGEhPKSWGU KU hGTGD[ C OCLQT EhCNÿ NGPIG. (KPCNN[, CU KP VhG RhCTOCEGWVKECN CPF KP VhG CTGC QH HQQF URQKNCIG [GCUVU, KV KU ETWEKCN VhCV GZRGTKOGPVCN TGUWNVU KP NCDQTCVQT[ UVTCKPU CPF UGVVKPIU CTG XCNKFCVGF KP VTWG URQKNCIG KUQNCVGU. 6hG DKQEhGOKECN VQQNU UWEh CU UVTWEVWTCN CPCN[UKU U[UVGOU HQT [GCUV EGNN YCNNU CPF OGODTCPGU, CPVKDQFKGU VQ UVTGUU RTQVGKPU CPF KP XKXQ CEVKXÿ KV[ CUUC[U QH CPVKOKETQDKCN TGUKUVCPEG OGEhCPKUOU CTG CXCKNCDNG. 2CTVKEWNCTN[ TGNGÿ XCPV KU VhG UVWF[ QH RTGUGTXCVKXG UVTGUU TGURQPUG KP VhG URGEKGU Zygosaccharomyces bailii CNUQ FKUEWUUGF KP 2KRGT GV CN. þú001). 5VKNN, VhKU KU C TGNCVKXGN[ NKVVNG GZRNQTGF URGEKGU YKVh TGURGEV VQ OQNGEWNCT KPUKIhV KP TGURQPUG VQ HQQF RTGUGTXCVKXGU YhKNG VhG TGNGXCPEG KU hKIh, UKPEG Z. bailii KU VhG OQTG TGEWTTKPI KPHGEVKQWU EGNN V[RG KP HQQF URQKNCIG KPEKFGPVU.
#EMPQYNGFIGOGPVU 9G YQWNF NKMG VQ VhCPM VhG OGODGTU QH VhG /KETQDKQNQI[ CPF VhG /QNGEWNCT $KQNÿ QI[ î /KETQDKCN (QQF 5CHGV[ üTQWR QH VhG 5YCOOGTFCO +PUVKVWVG HQT øKHG 5EKÿ GPEGU CPF QH VhG /KETQDKQNQI[ TGUGCTEh ITQWRU CV 6÷ò (QQF 4GUGCTEh HQT UVKOWÿ NCVKPI FKUEWUUKQPU. #V ðPKNGXGT 4GUGCTEh, YG YQWNF NKMG VQ VhCPM KP RCTVKEWNCT JQhP %hCROCP CPF /CNEQNO 5VTCVHQTF HQT KPURKTKPI FKUEWUUKQPU QXGT VhG NCUV GKIhV [GCTU. (KPCNN[ YG CEMPQYNGFIG VhG NQPI UVCPFKPI EQNNCDQTCVKQP YKVh VhG ITQWR QH RTQH. %. 6hGQ 8GTTKRU, RTQHGUUQT QH CRRNKGF OQNGEWNCT EGNN DKQNQI[ CV ðVTGEhV ðPKÿ XGTUKV[ CPF HQTOGT ITQWR NGCFGT $KQ6GEhPQNQI[ CPF ©ZRNQTCVQT[ 5EKGPEG CV ðPKNGÿ XGT 4GUGCTEh î &GXGNQROGPV. #PFTG $QQTUOC KU UWRRQTVGF D[ VhG (QWPFCVKQP HQT 6GEhPQNQIKECN 5EKGPEGU þ569).
üGHGTGPEGU #DGG 6 CPF 9QWVGTU J# þ1ýýý) /KETQDKCN UVTGUU TGURQPUG KP OKPKOCN RTQEGUUKPI. +PV J (QQF /KETQDKQN õ0:ñõÿý1
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#NYC\GGT &, %CEhQP 4 CPF &KXKGU %. þú00ú) $GhCXKQT QH Lactobacillus plantarum CPF Saccharomyces cerevisiae KP HTGUh CPF VhGTOCNN[ RTQEGUUGF QTCPIG LWKEG. J (QQF 2TQV ñõï1õôñÿ1õôý #ODGUK #, /KTCPFC /, 2GVTQX 88 CPF 5NC[OCP 9 þú000) $KQIGPGUKU CPF HWPEVKQP QH VhG [GCUV RNCUOCÿOGODTCPG *íÿ#62CUG. J ©ZR $KQN ú03 1õõÿ1ñ0 #PFTGYU 2& CPF 5VCTM /J þú000) &[PCOKE, 4hQ1RÿFGRGPFGPV NQECNK\CVKQP QH 2ME1R VQ UKVGU QH RQNCTK\GF ITQYVh. J %GNN 5EK 113ïúñôõÿúñý3 $CKTFÿ2CTMGT #%, $WEhCPCP 4ø, $WUVC ((, %QNG /$, &Q[NG, /2, ©[NGU /, (CTMCU J, (NQYÿ GTU 45, JQWXG Jø CPF /GPFQ\C 5 þ1ýýô) 5QHV FTKPMU, HTWKV LWKEGU, EQPEGPVTCVGU CPF HTWKV RTGUGTXGU. +P 4QDGTU 6#, 2KVV J+, (CTMCU J CPF üTCW (* þGFU) /KETQQTICPKUOU KP HQQFU ñïOKETQDKCN GEQNQI[ QH HQQF EQOOQFKVKGU. $NCEMKG #ECFGOKE î 2TQHHGUUKQPCN, øQPFQP, ðPKVGF -KPIFQO. RR. ùù0ÿùñ0 $CNGKTCU %QWVQ //, *CTVQI $J, *WKU KP •V 8GNF J*J, *QHUVTC * CPF XCP FGT 8QUUGP J/$/ þ1ýýñ) +FGPVKHKECVKQP QH URQKNCIG [GCUVU KP C HQQFÿRTQFWEVKQP EhCKP D[ OKETQUCVGNNKVG RQN[OGTCUG EhCKP TGCEVKQP HKPIGTRTKPVKPI. (QQF /KETQDKQN 13ïõýÿñ7 $QNGU © CPF *QNNGPDGTI %2 þ1ýý7) 6hG OQNGEWNCT IGPGVKEU QH hGZQUG VTCPURQTV KP [GCUVU. (©/5 /KETQDKQN 4GX ú1ïôõÿ111 $QO +J, -NKU (/, FG ÷QDGN * CPF $TWN 5 þú001) # PGY UVTCVGI[ HQT KPhKDKVKQP QH VhG URQKNÿ CIG [GCUV Saccharomyces cerevisiae CPF Zygosaccharomyces bailii DCUGF QP EQODKPCÿ VKQP QH C OGODTCPGÿCEVKXG RGRVKFG YKVh CP QNKIQUCEEhCTKFG VhCV NGCFU VQ CP KORCKTGF IN[EQU[NRhQURhCVKFWNKPQUKVQN þü2+)ÿFGRGPFGPV [GCUV YCNN RTQVGKP NC[GT. (©/5 ;GCUV 4GUGCTEh 1ï1ô7ÿ1ýù $QQV 4ü, 4GPMGOC ü*, 5VTKLNCPF #, XCP
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5. $TWN GV CN.
%CTQ ø*, 6GVVGNKP *, 8QUUGP J*, 4CO #(, XCP FGP ©PFG * CPF -NKU (/ þ1ýý7) +P UKNKEKQ KFGPVKHKECVKQP QH IN[EQU[NÿRhQURhCVKÿF[NKPQUKVQNÿCPEhQTGF RNCUOCÿOGODTCPG CPF EGNN YCNN RTQVGKPU QH Saccharomyces cerevisiae. ;GCUV 13ï1ù77ÿ1ùôý %hGPI ø, /QIhTCD[ J CPF 2KRGT 29 þ1ýýý) 9GCM QTICPKE CEKF VTGCVOGPV ECWUGU C VTGhCNQUG CEEWOWNCVKQP KP NQYÿR* EWNVWTGU QH Saccharomyces cerevisiae, PQV FKURNC[GF D[ VhG OQTG RTGUGTXCVKXGÿTGUKUVCPV Zygosaccharomyces bailii. (©/5 /KETQDKQN øGVV 170ïôýÿ ýõ %NGXGNCPF J, /QPVXKNNG 6J, ÷GU +(, %hKMKPFCU /ø þú001) $CEVGTKQEKPUû UCHG PCVWTCN CPVKOÿ KETQDKCNU HQT HQQF RTGUGTXCVKQP. +PV J (QQF /KETQDKQN 71ï1ÿú0 &CXKFUQP 2/ CPF *CTTKUQP /# þú00ú) 4GUKUVCPEG CPF CFCRVCVKQP VQ HQQF CPVKOKETQDKCNU, UCPKVK\GTU, CPF QVhGT RTQEGUU EQPVTQNU. (QQF 6GEhPQNQI[ õñïñýÿ7ô &GCM 6 þ1ýý1) (QQF DQTPG [GCUVU. #FX #RRN /KETQDKQN 3ýï17ý ú7ô &G $QGT © CPF ÷KGNUGP 28 þ1ýýõ) (QQF RTGUGTXCVKXGU. +P 4.#. 5COUQP, ©.5. *QGMUVTC, J.%. (TKUXCF CPF ò. (KNVGPDQTI þGFU), +PVTQFWEVKQP QH HQQFÿDQTPG HWPIK. %GPVTCCNDWTGCW XQQT UEhKOOGNEWNVWTGU, 9CIGPKPIGP, 6hG ÷GVhGTNCPFU. RR. úôýÿúý3 &G ÷QDGN Jü , -NKU (/, 2TKGO J, /WPPKM 6 CPF XCP FGP ©PFG * þ1ýý0) 6hG INWECPCUGÿ UQNWDNG OCPPQRTQVGKPU NKOKV EGNN YCNN RQTQUKV[ KP Saccharomyces cerevisiae. ;GCUV ñïùý1ÿùýý. &G ÷QDGN *, 4WK\ %, /CTVKP *, /QTTKU 9, $TWN 5, /QNKPC / CPF -NKU (/ þú000) %GNN YCNN RGTVWTDCVKQP KP [GCUV TGUWNVU KP FWCN RhQURhQT[NCVKQP QH VhG 5NVúö/RM1 /#2 MKPCUG CPF KP CP 5NVúÿOGFKCVGF KPETGCUG KP FKS2-lacZ GZRTGUUKQP, INWECPCUG TGUKUVCPEG CPF VhGTÿ OQVQNGTCPEG. /KETQDKQNQI[ 1ùñïú1ú1ÿú13ú &G ÷QDGN *,5KGVUOC J*, XCP FGP ©PFG * CPF -NKU (/ þú001) /QNGEWNCT QTICPK\CVKQP CPF EQPUVTWEVKQP QH VhG HWPICN EGNN YCNN. +Pï6hG /[EQVC 8+++ $KQNQI[ QH VhG (WPICN %GNN þ*QYCTF 4J CPF üQY ÷#4, ©FU.) 5RTKPIGT 8GTNCI,$GTNKP &G4KUK Jø, +[GT 84, $TQYP 2ò. þ1ýý7) ©ZRNQTKPI VhG OGVCDQNKE CPF IGPGVKE EQPVTQN QH IGPG GZRTGUUKQP QP C IGPQOKE UECNG. 5EKGPEG úùïñô0ÿñôñ &KGFGTKEh J#, 5EhGRRGT /, XCP *QGM 2, øWVVKM /#*, XCP &KLMGP J2, 2TQPM J6, -NCCUUGP 2, $QGNGPU *(/, 6GKZGKTC FG /CVVQU /J, XCP &CO - CPF -TWEMDGTI #ø þ1ýýý) üNWEQUG WRVCMG MKPGVKEU CPF VTCPUETKRVKQP QH *:6 IGPGU KP EhGOQUVCV EWNVWTGU QH Saccharomyces cerevisiae. J $KQN %hGO ú7ùï1õ3õ0ÿ1õ3õý &KLMITCCH üJ, $TQYP Jø CPF $WUUG[ * þ1ýýñ) 6hG -÷*1 IGPG QH Saccharomyces cerevisiae KU C HWPEVKQPCN hQOQNQIWG QH -4©ý. ;GCUV 1úïñô3ÿñýú &QP\GCW /, $QWTFKPGCWF J2 CPF øCWSWKP üJ þ1ýýñ) 4GIWNCVKQP D[ NQY VGORGTCVWTGU CPF CPCGTQDKQUKU QH C [GCUV IGPG URGEKH[KPI C RWVCVKXG ü2+ÿCPEhQTGF RNCUOC OGODTCPG RTQVGKP. /QN /KETQDKQN ú0ïùùýÿùõý &QWINCU %/, &•+RRQNGVQ J#, 5hGK üJ, /GKP\ /, òPKUhK J, /CTTKPCP J#, øK 9, #DTW\\Q ü-, (NCVVGT[ #, $CTVK\CN -, /KVEhGNN # CPF -WTV\ /$ þ1ýý7) +FGPVKHKECVKQP QH VhG (-51 IGPG QH Candida albicans CU VhG GUUGPVKCN VCTIGV QH 1,3DGVCÿ&ÿINWECP U[PVhCUG KPhKDKVQTU. #PVKOKETQD #IGPVU %hGOQVhGT ù1ïúù71ÿúù7ý ©KUGP /$, 5RGNNOCP 26, $TQYP 2ò, $QVUVGKP & þ1ýýô) %NWUVGT CPCN[UKU CPF FKURNC[ QH IGÿ PQOGÿYKFG GZRTGUUKQP RCVVGTPU. 2TQE ÷CVN #ECF 5EK ð5# úõï1ùôñ3ÿ1ùôñô (NGGV ü þ1ýýú) 5RQKNCIG [GCUVU. %TKV 4GX $KQVGEhPQN 1úï1ÿùù üKCGXGT ü, %hW #/, ÷K ø, %QPPGNN[ %, 4KNGU ø, 8GTQPPGCW 5, &QY 5, øWECWÿ&CPKNC #, #PFGTUQP -, #PFTG $, #TMKP #2, #UVTQOQHH #, ©Nÿ$CMMQWT[ /, $CPIhCO 4, $GPKVQ 4, $TCEhCV 5, %CORCPCTQ 5, %WTVKUU /, &CXKU -, &GWVUEhDCWGT #, ©PVKCP -&, (NChGTV[ 2, (QWT[ (, üCTHKPMGN &J, üGTUVGKP /, üQVVG &, üWNFGPGT ð, *GIGOCPP J*, *GORGN 5, *GTOCP <, JCTCOKNNQ &(, -GNN[ &©, -GNN[ 5ø, -QVVGT 2, øC$QPVG &, øCOD &%, øCP ÷,
ý ;GCUVU CPF HQQF URQKNCIG
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øKCPI *, øKCQ *, øKW ø, øWQ %, øWUUKGT /, /CQ 4, /GPCTF 2, òQK 5ø, 4GXWGNVC Jø, 4QDGTVU %J, 4QUG /, 4QUUÿ/CEFQPCNF 2, 5EhGTGPU $, 5EhKOOCEM ü, 5hCHGT $, 5hQGÿ OCMGT &&, 5QQMhCKÿ/ChCFGQ 5, 5VQTOU 4-, 5VTCVhGTP J÷, 8CNNG ü, 8QGV /, 8QNÿ EMCGTV ü, 9CPI %;, 9CTF 64, 9KNhGNO[ J, 9KP\GNGT ©#, ;CPI ;, ;GP ü, ;QWPIOCP ©, ;W -, $WUUG[ *, $QGMG J&, 5P[FGT /, 2hKNKRRUGP 2, &CXKU 49, JQhPUVQP /. þú00ú) (WPEVKQPCN RTQHKNKPI QH VhG Saccharomyces cerevisiae IGPQOG. ÷CVWTG úõï3ô7ÿ 3ý1 üQNFOCP 4%, 5WNNKXCP 2#,
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5. $TWN GV CN.
/CPCXCVhW ©-, &KOOQEM J4, 8CUhKUVhC 5% CPF %hCPFTCUGMCT 2* þú001) +PhKDKVKQP QH *þí)ÿ#62CUGÿOGFKCVGF RTQVQP RWORKPI KP Cryptococcus neoformans D[ C PQXGN EQPÿ LWICVGF UV[T[N MGVQPG. J #PVKOKETQD %hGOQVhGT ù7ïùý1ÿùýù /CTSWCTFV * þ1ýñú) > (GKPDCW XQP *GHG\GNNGP KO ©NGMVTQPGPOKMTQUMQR ++ /KVVïSaccharomyces cerevisiae 5VkOOG. < ÷CVWTHQTUEh 17ïñôýÿñýõ /CT\C ©, %COQWITCPF ÷ CPF /CPQP 5 þú00ú) $CZ GZRTGUUKQP RTQVGEVU [GCUV RNCUOC OGOÿ DTCPG CICKPUV GVhCPQNÿKPFWEGF RGTOGCDKNKUCVKQP. (©$5 øGVV õú1ïù7ÿõú /C\WT 2, /QTKP ÷, $CIKPUM[ 9, GNÿ5hCTDKPK /, %NGOCU J#, ÷KGNUGP J$ CPF (QQT ( þ1ýýõ) &KHHGTGPVKCN GZRTGUUKQP CPF HWPEVKQP QH VYQ hQOQNQIQWU UWDWPKVU QH [GCUV 1,3ÿDGVCÿ&ÿ INWECP U[PVhCUG. /QN %GNN $KQN 1õïõñ71ÿõñô1 /GODTG J/, -WDCE\MC / CPF %hGPG % þ1ýýý) %QODKPGF GHHGEVU QH R* CPF UWICT QP ITQYVh TCVG QH Zygosaccharomyces rouxii, C DCMGT[ RTQFWEV URQKNCIG [GCUV. #RRN ©PX /KETQDKQN ñõïùýú1ÿùýúõ /GPUQPKFGU (+%, 5EhWWTOCPU /J, 6GKZGKTC FG /CVVQU J, *GNNKPIYGTH -J CPF $TWN 5 þú00ú) 6hG OGVCDQNKE TGURQPUG QH Saccharomyces cerevisiae VQ EQPVKPWQWU hGCV UVTGUU. /QN $KQN 4GRQTVU úýï103ÿ10ñ /QNNCRQWT /, 2KRGT 29 þú001) 6hG
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òXCNNG 4, øKO 56, *QNFGT $, JWG %-, /QQTG %9 CPF øKRMG 2÷ þ1ýýô) # URhGTQRNCUV TCVG CUUC[ HQT FGVGTOKPCVKQP QH EGNN YCNN KPVGITKV[ KP [GCUV. ;GCUV 1ùï11õýÿ11ññ ò\ECP 5 CPF JQhPUVQP / þ1ýýý) (WPEVKQP CPF TGIWNCVKQP QH [GCUV hGZQUG VTCPURQTVGTU. /Kÿ ETQDKQN /QN $KQN 4GX ñ3ïõõùÿõñý 2CPCTGVQW $ CPF 2KRGT 29 þ1ýýú) 6hG RNCUOC OGODTCPG QH [GCUV CESWKTGU C PQXGN hGCVÿ UhQEM RTQVGKP þhUR 30) CPF FKURNC[U C FGENKPG KP RTQVQPÿRWORKPI #62CUG NGXGNU KP TGÿ URQPUG VQ DQVh hGCV UhQEM CPF VhG GPVT[ VQ UVCVKQPCT[ RhCUG. ©WT J $KQEhGO ú0ñïñ3õÿ ñù0 2CVVKUQP 6ø CPF XQP *QN[ # þú001) ©HHGEV QH UGNGEVGF PCVWTCN CPVKOKETQDKCNU QP $CMGT•U [GCUV CEVKXKV[. øGVV #RRN /KETQDKQN 33ïú11ÿú1õ 2KRGT 2, òTVK\ÿ%CNFGTQP %, *QN[QCM %, %QQVG 2 CPF %QNG / þ1ýý7) *UR30, VhG KPVGITCN RNCUOC OGODTCPG hGCV UhQEM RTQVGKP QH Saccharomyces cerevsiae, KU C UVTGUUÿKPFWEKDNG TGIWNCVQT QH RNCUOC OGODTCPG *íÿ#62CUG. %GNNU 5VTGUU %hCRGTQPGU úï1úÿúù 2KRGT 2, /ChG ;, 6hQORUQP 5, 2CPFLCKVCP 4, *QN[QCM %, ©IPGT 4, /WhNDCWGT /, %QQVG 2 CPF -WEhNGT - þ1ýýô) 6hG 2FT1ú #$% VTCPURQTVGT KU TGSWKTGF HQT VhG FGXGNQROGPV QH YGCM QTICPKE CEKF TGUKUVCPEG KP [GCUV. ©/$ò J 17ïùúõ7ÿùúñõ 2KRGT 2, òTVK\ %, *CV\KZCPVhKU - CPF /QNNCRQWT / þú001) 9GCM CEKF CFCRVCVKQPû VhG UVTGUU TGURQPUG VhCV EQPHGTU [GCUVU YKVh TGUKUVCPEG VQ QTICPKE CEKF HQQF RTGUGTXCVKXGU. /KETQÿ DKQNQI[ 1ù7ïúñ3õÿúñùú 2QRQNQ ø CPF 8CK / þ1ýýý) 6hG üCU1 IN[EQRTQVGKP,C RWVCVKXG YCNN RQN[OGT ETQUUÿNKPMGT. $KQEhKO $KQRh[U #EVC 1ùúñï3ôõÿù00 2QRQNQ ø, üWCNVKGTK 6 CPF 4CIPK © þú001) 6hG [GCUV EGNNÿYCNN UCNXCIG RCVhYC[. /GF /[EQN 3ý þ5WRRN)ï111ÿ1ú1 4CLCXGN /, 2hKNKR $, $WGhTGT $/, ©TTGFG $ CPF øGXKP &© þ1ýýý) /KFú KU C RWVCVKXG UGPUQT HQT EGNN KPVGITKV[ UKIPCNKPI KP Saccharomyces cerevisiae. /QN %GNN $KQN 1ýï3ýñýÿ3ý7ñ 4GKHGPDGTI ©, $QNGU © CPF %KTKCE[ / þ1ýý7) -KPGVKE EhCTCEVGTKUCVKQP QH KPFKXKFWCN hGZQUG VTCPURQTVGTU QH Saccharomyces cerevisiae CPF VhGKT TGNCVKQP VQ VhG VTKIGTTKPI OGEhCÿ PKUOU QH INWEQUG TGRTGUUKQP. ©WT J $KQEhGO úùõï3úùÿ333 4GUV XCP FGT /©, -COOKPIC #*, ÷CMCPQ #, #PTCMW ;, 2QQNOCP $ CPF -QPKPIU 9÷ þ1ýýõ) 6hG RNCUOCOGODTCPG QH Saccharomyces cerevisiaeïUVTWEVWTG, HWPEVKQP, CPF DKQIGPGUKU. /KETQDKQN 4GX õýï30ùÿ3úú 4QFTKIWGU (, %QTVGÿ4GCN /, øGCQ %, XCP &KLMGP J2 CPF 2TQPM J6 þú001) òZ[IGP TGSWKTGÿ OGPVU QH VhG HQQF URQKNCIG [GCUV Zygosaccharomyces bailii KP U[PVhGVKE CPF EQORNGZ OGFKC. #RRN ©PXKTQP /KETQDKQN ñ7ïú1ú3ÿú1úô 4QNNGT 5 CPF %QXKNN ÷ þ1ýýý) 6hG CPVKHWPICN RTQRGTVKGU QH EhKVQUCP KP NCDQTCVQT[ OGFKC CPF CRRNG LWKEG. +PV J (QQF /KETQDKQN ù7ïñ7ÿ77 5CPINCTF & CPF òFFU (% þú00ú) 4GUKUVCPEG QH %CPFKFC URGEKGU VQ CPVKHWPICN CIGPVUïOQÿ NGEWNCT OGEhCPKUOU CPF ENKPKECN EQPUGSWGPEGU. øCPEGV +PHGEV &KU úï73ÿôõ 5hChKPKCP 5 CPF $WUUG[ * þú000) $GVCÿ1,ñÿINWECP U[PVhGUKU KP Saccharomyces cerevisiae. /QN /KETQDKQN 3õïù77ÿùôý 5hGCTGT #©*, /C\\QVVC #5, %hW[CVG 4 CPF üQODCU &© þú00ú) *GCV 4GUKUVCPEG QH JWKEG 5RQKNCIG /KETQQTICPKUOU. J (QQF 2TQV ñõï1ú71ÿ1ú7õ 5OGNV J22/, *GNNGOQPU J%, CPF 2CVVGTUQP / þú00úC)©HHGEVU QH hKIh RTGUUWTG QP XGIGVCVKXG OKETQQTICPKUOU. KP ðNVTC *KIh 2TGUUWTG 6TGCVOGPV QH (QQFU, *GPFTKEMZ /©ü CPF -PQTT & þGFU) -NWYGT #ECFGOKE, ÷;. þKP RTGUU) 5OGNV J22/, *GNNGOQPU J CPF $TWN 5 þú00úD) 2h[UKQNQIKECN CURGEVU QH 2TGUUWTG FGEQPÿ VCOKPCVKQP KP DWKNFKPI KPCEVKXCVKQP OQFGNU. KP 6TGPFU KP *KIh 2TGUUWTG $KQUEKGPEG î $KQVGEhPQNQI[, *C[CUhK, * þGF) ©NUGXKGT, ÷GY ;QTM RR. ùô7ÿùýñ.
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5. $TWN GV CN.
5VTCVHQTF /, $QPF %J, JCOGU 5#, 4QDGTVU ÷ CPF 5VGGNU * þú00ú) Candida davenporti UR. PQX., C RQVGPVKCN UQHVÿFTKPMU URQKNCIG [GCUV KUQNCVGF HTQO C YCUR. +PV J 5[UV ©XQN /Kÿ ETQDKQN õúï13ñýÿ137õ 5VGGIÿVGT 2(, òVVGP ü&, #NFGTNKGUVGP /, FG 9GKLGT 4, ÷CCMVIGDQTGP ü, $KLN J, 8CUDKPFGT #J, -GTUhQH +, CPF XCP &WKLXGPFKLM #/ þú001) /QFGNNKPI VhG GHHGEVU QH þITGGP) CPVKÿ HWPICNU, FTQRNGV UK\G FKUVTKDWVKQP CPF VGORGTCVWTG QP OQWNF QWVITQYVh KP YCVGTÿKPÿQKN GOWNUKQPU. +PV J (QQF /KETQDKQN ñ7ïúú7ÿú3ý 5VGGNU *, JCOGU 5#, 4QDGTVU +÷ CPF 5VTCVHQTF / þ1ýýý) <[IQUCEEhCTQO[EGU NGPVWUïC UKIÿ PKHKECPV PGY QUOQRhKNKE, RTGUGTXCVKXG TGUKUVCPV URQKNCIG [GCUV, ECRCDNG QH ITQYVh CV NQY VGORGTCVWTG. J #RRN /KETQDKQN ô7ïõú0ÿõú7 5VTCVHQTF / CPF #PUNQY 2# þ1ýýô) ©XKFGPEG VhCV UQTDKE CEKF FQGU PQV KPhKDKV [GCUV CU C ENCUUKECN YGCM CEKF ÂYGCM CEKF RTGUGTXCVKXG•. øGVV #RRN /KETQDKQN ú7ïú03ÿú0ñ 6CMGhKMQ 5ChCTC, 6CMCMQ üQFC, CPF 5CVQTW òhIK[C þú00ú) %QORTGhGPUKXG GZRTGUUKQP CPCN[UKU QH VKOGÿFGRGPFGPV IGPGVKE TGURQPUGU KP [GCUV EGNNU VQ NQY VGORGTCVWTG. J. $KQN. %hGO. ú77ïõ001õÿõ00ú1 6hQORUQP J4, &QWINCU %/, øK 9, JWG %-, 2TCOCPKM $, ;WCP :, 4WFG 6*, 6QHHCNGVVK &ø, 2GTHGEV J4 CPF -WTV\ / þ1ýýý) # INWECP U[PVhCUG FKS1 hQOQNQIWG KP Cryptococcus neoformans KU UKPING EQR[ CPF GPEQFGU CP GUUGPVKCN HWPEVKQP. J $CEVGTKQN 1ô1ïùùùÿùõ3 6QhÿG #, ;CUWPCIC 5, ÷KUQIK *, 6CPCMC -, òIWEhK 6 CPF /CVUWK ; þ1ýý3)6hTGG [GCUV IGPGU,2+41 ,2+4ú CPF 2+43 ,EQPÿVCKPKPI KPVGTPCN VCPFGO TGRGCVU,CTG TGNCVGF VQ GCEh QVhGT,CPF 2+41 CPF 2+4ú CTG TGSWKTGF HQT VQNGTCPEG VQ hGCV UhQEM. ;GCUV ýïùô1ÿùýù 6QMWQMC - þ1ýý3) 5WICTÿ CPF UCNVÿVQNGTCPV [GCUVU. J #RRN $CEVGTKQN 7ùï101ÿ110 8CNFKXKGUQ /*, (GTTCTKQ ø, 8CK /, &WTCP # CPF 2QRQNQ ø þú000) %hKVKP U[PVhGUKU KP C ICU1 OWVCPV QH 5CEEhCTQO[EGU EGTGXKUKCG. J $CEVGTKQN 1ôúïù7õúÿù7õ7 8CP FGT 8QUUGP J/$/, 4ChCQWK *, &G ÷KLU /9%/ CPF *CTVQI $J þú003) 2%4 OGVhQFU HQT VTCEKPI CPF FGVGEVKQP QH [GCUVU KP VhG HQQF EhCKP. +P $QGMhQWV 6 CPF 4QDGTV 8 þGFU) ;GCUVU, $GhT•U 8GTNCI üOD* î %Q, *CODWTI, üGTOCP[ þ+P 2TGUU) 8CUEQPEGNNGU, /. J., JKCPI, ;., /E&CKF, -., üKNQQN[, ø., 9TGV\GN, 5., 2QTVGT, &. ø., /CTVKP, %. ©., CPF üQNFDGTI, /. #. þú001) +FGPVKHKECVKQP CPF EhCTCEVGTK\CVKQP QH C NQY QZ[IGP TGURQPUG GNGOGPV KPXQNXGF KP VhG h[RQZKE KPFWEVKQP QH C HCOKN[ QH Saccharomyces cerevisiae IGPGU. +ORNKECVKQPU HQT VhG EQPUGTXCVKQP QH QZ[IGP UGPUKPI KP GWMCT[QVGU. J $KQN %hGO ú7ñ, 1ù37ùÿ1ù3ôù 8GTPC J, øQFFGT #, øGG -, 8CIVU # CPF $CNNGUVGT 4 þ1ýý7) # HCOKN[ QH IGPGU TGSWKTGF HQT OCKPVGPCPEG QH EGNN YCNN KPVGITKV[ CPF HQT VhG UVTGUU TGURQPUG KP Saccharomyces cerevisiae. 2TQE ÷CVN #ECF 5EK ð5# ýùï13ô0ùÿ13ô0ý 8GTYCCN 4, 2CCNOCP J9, *QIGPMCOR #, 8GTMNGKL #, 8GTTKRU %6 CPF $QQPUVTC J þú00ú) *:6õ GZRTGUUKQP KU FGVGTOKPGF D[ ITQYVh TCVGU KP Saccharomyces cerevisiae. ;GCUV 1ýï10úýÿ103ô 8KEGPVG /(, %CDGNNQ #, 2NCVCU ü, $CUKNKQ #, &KG\ /6, &TGKMQTP 5, üKCEQDDG 4#, òPKUhK J%, /GKP\ /, -WTV\ /$, 4QUGPDCEh /, 6hQORUQP J, #DTW\\Q ü, (NCVVGT[ #, -QPI ø, 6UKRQWTCU #, 9KNUQP -©, 2GNCG\ ( þú001) #PVKOKETQDKCN CEVKXKV[ QH GTIQMQPKP # HTQO 6TKEhQFGTOC NQPIKDTCEhKCVWO . J #RRN /KETQDKQN ý1ïô0ñÿô13 8QU 9/ÿFG þú001) #FXCPEGU KP IGPQOKEU HQT OKETQDKCN HQQF HGTOGPVCVKQPU CPF UCHGV[. %WTT òRKP $KQVGEhPQN 1úïùý3ÿùýô 9CPKUMC 4&, 8GPMCVhGUC 46, %hCPFTCUhGMCT #, -TKUhPCXGPK 5, $GLQUCPQ (2, JGQWPI J, JC[CTCL, /WVhWMTKUhPCP 5 CPF øKCPI ü* þú001) #PVKHWPICN RTQVGKPU CPF QVhGT OGEhCÿ PKUOU KP VhG EQPVTQN QH UQTIhWO UVCNM TQV CPF ITCKP OQNF. J #ITKEWNV (QQF %hGO ùýïù73úÿù7ùú
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9KGE\QTMG 4, -TCORG 5, 9GKGTUVCNN 6, (TGKFGN -, *QNNGPDGTI %2 CPF $QNGU © þ1ýýý) %QPÿ EWTTGPV MPQEMÿQWV QH CV NGCUV ú0 VTCPURQTVGT IGPGU KU TGSWKTGF VQ DNQEM WRVCMG WRVCMG QH hGZQUGU KP 5CEEhCTQO[EGU. (©$5 øGVV ùñùï1ú3ÿ1úô 9QWVGTU 2%, #NXCTG\ + CPF 4CUQ J þú001C) %TKVKECN HCEVQTU FGVGTOKPKPI KPCEVKXCVKQP MKPGVKEU D[ RWNUGF GNGEVTKE HQQF RTQEGUUKPI. 6TGPFU (QQF 5EK 6GEhPQN 1úï11úÿ1ú1 9QWVGTU 2%, $QU #2 CPF ðGEMGTV J þú001D) /GODTCPG RGTOGCDKNK\CVKQP KP TGNCVKQP VQ KPCEÿ VKXCVKQP MKPGVKEU QH øCEVQDCEKNNWU URGEKGU FWG VQ RWNUGF GNGEVTKE HKGNFU. #RRN ©PX /Kÿ ETQDKQN ñ7ï30ýúÿ3101 <W 6, 8GTPC J CPF $CNNGUVGT 4 þú001) /WVCVKQPU KP 95% IGPGU HQT RWVCVKXG UVTGUU TGEGRVQTU TGUWNV KP UGPUKVKXKV[ VQ OWNVKRNG UVTGUU EQPFKVKQPU CPF KORCKTOGPV QH 4NO1ÿFGRGPFGPV IGPG GZRTGUUKQP KP Saccharomyces cerevisiae . /QN üGPGV üGPQOKEU úññï1ùúÿ1õõ
10 NQP-EQPXGPVKQPCl [GCUVU KP CPVKHWPICl CRRlKECVKQP 8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
#DUVTCEV 6JG CPVKOKETQDKCNÿ CPF JGPEG CPVKHWPICN GHHGEVU QH [GCUV-RTQFWEGF GVJCPQN JCU DGGP TGEQIPKUGF HQT EGPVWTKGU. #RRNKECVKQP QH [GCUVU HQT DKQEQPVTQN QH HWPIK KP CITKEWNVWTGÿ RQUVJCTXGUV VGEJPQNQI[ÿ HQTGUV KPFWUVT[ÿ CPF HQQF UEKGPEG RTGUGPVU C PGY CTGC QH DKQVGEJPQNQI[. $KQEQPVTQN GPXKTQPOGPVU CTG EQORNGZ; EQPUGSWGPVN[ÿ VJGTG KU CP GPQTOQWU FKXGTUKV[ COQPI VJG QTICPKUOU KPXQNXGF KP DKQEQPVTQN CPF QH KPJKDKVKQP OGEJCPKUOU. %WTTGPV IGPGVKE CRRTQCEJGU HQEWU OCKPN[ QP VJG WUG QH OQNGEWNCT OGVJQFU HQT VJG KFGPVKHKECVKQP CPF RJ[NQIGPGVKE CPCN[UKU QH DKQEQPVTQN QTICPKUOU. )TQYKPI KPHQTOCVKQP KU CXCKNCDNG CDQWV VJG IGPGVKE U[UVGOU QH C HGY DKQEQPVTQN [GCUVUÿ KPENWFKPI UGZWCNKV[ÿ EJTQOQUQOG PWODGTÿ RNQKF[ NGXGNÿ GZVTCEJTQOQUQOCN GNGOGPVUÿ CPF IGPGVKEU QH MKNNGT EJCTCEVGTU. (GY IGPGU JCXG DGGP ENQPGFÿ VQ FCVG OCKPN[ T&0# IGPGU. /GVJQFU HQT IGPGVKE OCPKRWNCVKQP JCXG DGGP FGUETKDGF HQT DKQEQPVTQN [GCUVU NKMG Aureobasidium pullulansÿ Pseudozyma flocculosaÿ Rhodosporidium toruloidesÿ Candida oleophilaÿ Pichia anomalaÿ CPF Pichia guilliermondii. (KTUV GZCORNGU QH VJG CRRNKECVKQP QH IGPGVKECNN[ GPIKPGGTGF UVTCKPU KP DKQEQPVTQN VTKCNU JCXG TGEGPVN[ DGEQOG CXCKNCDNG.
10.1 +PVTQFWEVKQP þGCUVU JCXG VQ KPVGTCEV YKVJ QVJGT OKETQQTICPKUOU KP VJGKT JCDKVCVU ý(NGGV 199ü; (NGGV 1999û. 0GICVKXG QT KPJKDKVKPI KPVGTCEVKQPU QH [GCUVU YKVJ QVJGT HWPIK ýKPENWFKPI [GCUVUû HTGSWGPVN[ QEEWTÿ KPENWFKPI EQORGVKVKQPÿ CPVCIQPKUO QT COGPUCNKUOÿ RCTCUKVKUO QT RTGFCVKQP ý$QFF[ CPF úKORGPP[ 199ù; (NGGV 199ü; (NGGV 1999û. 5EKGPVKHKE KPVGTGUV VQ WVKNK\G VJKU CPVKHWPICN CEVKXKV[ HQT VJG DKQEQPVTQN QH WPYCPVGF HWPIK CTQUG SWKVG GCTN[ ýþCTYQQF 193ùû. *QYGXGTÿ VJG QNFGUV CRRNKGF CPVKOKETQDKCN CEVKXKV[ QH [GCUVU KU VJG RTQFWEVKQP QH GVJCPQN. &WTKPI PCVWTCN YKPG HGTOGPVCVKQPÿ HKNCOGPVQWU HWPIK CPF DCEVGTKC DGEQOG KPJKDKVGF D[ VJG RTQFWEGF GVJCPQN. 5WDUGSWGPVN[ÿ OQUV QH VJG PQP-Saccharomyces [GCUVU VJCV FQOKPCVG KP VJG DGIKPPKPI QH VJG RTQEGUU CTG TGRNCEGF D[ Saccharomyces YKVJ KPETGCUKPI GVJCPQN EQPEGPVTCVKQPU ý©INK GV CN. 199ü; /KNNU GV CN. ùøøù; UGG CNUQ %JCRVGT 4ÿ RCTCITCRJ 4.ù.1û. #PQVJGT KORQTVCPV CPVKHWPICN CEVKXKV[ KU VJG EQORGVKVKQP HQT GUUGPVKCN QTICPKE CPF KPQTICPKE EQORQWPFU CPF HQT URCEG. #V VJG DGIKPPKPI QH YKPG HGTOGPVCVKQPÿ VJG TGURKTKPI QTICPKUOU EQORGVG HQT VJG TGUKFWCN QZ[IGP. #U C TGUWNVÿ VJG QZ[IGP KU EQPUWOGF CPF VJG EWNVWTG DGEQOGU CNOQUV CPCGTQDKEÿ YJKEJ KPJKDKVU VJG ITQYVJ QH CGTQDKE HWPIK Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
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8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
ý*CPUGP GV CN. ùøø1û. +V KU C YGNN-MPQYP RJGPQOGPQP VJCV HTWKVU DGEQOG OQTG UWUEGRVKDNG VQ TQV CHVGT YCUJKPI. 6JG PCVWTCN GRKRJ[VKE OKETQHNQTC QP VJG RJ[NNQRNCPG CPF HTWKVUÿ EQPUKUVKPI OCKPN[ QH DCEVGTKC CPF [GCUVUÿ YCU UJQYP VQ RTGXGPV VJG ITQYVJ QH HWPICN RCVJQIGPU ý$NCMGOCP CPF (QMMGOC 19üù; &TQD[ GV CN. 199ùû. (WTVJGT CPVKHWPICN CEVKXKVKGU QH [GCUVU KPENWFG VJG RTQFWEVKQP QH VQZKE GZVTCEGNNWNCT EQORQPGPVU NKMG MKNNGT RTQVGKPU ý/CTSWKPC GV CN. ùøøù; 5EJOKVV CPF $TGKPKI ùøøùûÿ EGNN YCNN FGITCFKPI GP\[OGU ý%CUVQTKC GV CN. 1997û QT QVJGT CPVKDKQVKE OGVCDQNKVGU ý#XKU CPF $oNCPIGT ùøøùûÿ UVKOWNCVKQP QH JQUV FGHGPEG TGURQPUGU ýúKNUQP GV CN. 1994ûÿ O[EQRCTCUKVKUO ý©NCF GV CN. 199÷ûÿ CPF RTGFCVKQP ýLCEJCPEG GV CN. ùøøøû. +P CFFKVKQP VQ VJG CDQXG-OGPVKQPGF RTQFWEVKQP QH CNEQJQNKE DGXGTCIGUÿ [GCUVU YKVJ CPVKHWPICN CEVKXKV[ JCXG CP KPETGCUKPI KORCEV KP DKQVGEJPQNQI[. &WG VQ GPXKTQPOGPVCN CPF JGCNVJ EQPEGTPU CDQWV VJG WUG QH HWPIKEKFGU KP RNCPV RTQVGEVKQP CPF RQUVJCTXGUV RTGUGTXCVKQPÿ VJGTG KU C UVTQPI FGOCPF HQT CNVGTPCVKXG OGCPU VJCV RTGXGPV VJG FGXGNQROGPV QH RCVJQIGPKE HWPIK. $KQRTQVGEVKQP QH RNCPVU CPF DKQEQPUGTXCVKQP KP VJG RQUVJCTXGUV VTGCVOGPV D[ [GCUVU ECP RTQXKFG UWEJ CP CNVGTPCVKXG ý$NCMGOCP CPF (QMMGOC 19üù; 2C[PG GV CN. ùøøø; &TWXGHQTU GV CN. ùøøù; ,CPKUKGYKE\ CPF öQTUVGP ùøøùû. 6YQ RTQFWEVU DCUGF QP VJG [GCUVU Candida oleophila CPF Cryptococcus albidus CTG EWTTGPVN[ EQOOGTEKCNN[ CXCKNCDNG HQT VJG EQPVTQN QH OQWNF FGEC[ KP EKVTWUÿ CRRNGUÿ CPF RGCTU ý#URKTG®ÿ ©EQIGP +PE. LCPIJQTPÿ 2#; þKGNFRNWU®ÿ #PEJQT þGCUVÿ %CRG 6QYPÿ 5QWVJ #HTKECû. 6JG [GCUV-NKMG DCUKFKQO[EGVQWU HWPIWU Ampelomyces quisqualis JCU DGGP HQTOWNCVGF CPF EQOOGTEKCNKUGF CU #õ-1ø® D[ ©EQIGP +PE. HQT VJG EQPVTQN QH RQYFGT[ OKNFGY KP ITGGPJQWUGU. 2NCPV 2TQFWEVU %Q. LVF ý$TCORVQPÿ %CPCFCû JCU TGIKUVGTGF C RTQFWEVÿ 5RQTQFGZ®ÿ YJGTG EQPKFKC QH VJG DCUKFGQO[EGVQWU [GCUV-NKMG HWPIWU Pseudozyma flocculosa CEV CU DKQHWPIKEKFG ý2CWNKV\ CPF $oNCPIGT ùøø1û. 6JG MKNNGT RJGPQOGPQP JCU DGGP CRRNKGF HQT VCZQPQOKECN RWTRQUGU CPF OC[ DG WUGHWN HQT VJG HGGF CPF HQQF KPFWUVT[ CPF KP OGFKEKPG ý/CINKCPK GV CN. 1997D; /CTSWKPC GV CN. ùøøùû. 5QOG UWIIGUVGF CPVKHWPICN [GCUVU ECP WPFGT EGTVCKP EKTEWOUVCPEGU CNUQ CEV CU URQKNCIG [GCUVUÿ ECWUKPI EQPUKFGTCDNG NQUUGU KP HGGF CPF HQQF KPFWUVT[ ý(NGGV 199ùû. #U FKUEWUUGF CDQXGÿ XCTKQWU OGEJCPKUOU ECP RTQXKFG CPVKHWPICN CEVKXKV[. /QTGQXGTÿ VJGTG KU C JWIG CPF KPETGCUKPI PWODGT QH FQEWOGPVGF [GCUVU YKVJ UWEJ CEVKXKVKGU ý2CWNKV\ CPF $oNCPIGT ùøø1; ,CPKUKGYKE\ CPF öQTUVGP ùøøù; /CTSWKPC GV CN. ùøøùû. 6JGTGHQTGÿ OCP[ FKHHGTGPV IGPGVKE HCEVQTU CTG KPXQNXGF KP VJG TGURGEVKXG CPVCIQPKUVKE DGJCXKQWT. (QT KPUVCPEGÿ UGXGTCN IGPGU CPF TGIWNCVQT[ PGVYQTMU FGVGTOKPG GVJCPQN VQNGTCPEG KP [GCUVU ý,QPGU 19ü9; 2KRGT 199ôû. 5GXGTCN RJ[UKQNQIKECN UVTCVGIKGU JCXG DGGP GXQNXGF KP [GCUVU HQT JCPFNKPI QZ[IGP NKOKVCVKQPÿ CPF VJG GZRTGUUKQP QH OCP[ IGPGU KU TGIWNCVGF D[ OWNVKRNG cis CPF trans CEVKPI GNGOGPVU ý2Q[VQP 1999; 6GT LKPFG CPF 5VGGPUOC ùøøù; 2CUUQVJ GV CN. ùøø3û. /CP[ IGPGU CTG KPXQNXGF KP VJG EQORGVKVKQP HQT MG[ PWVTKGPVU UWEJ CU UWICTU ý$QNGU CPF *QNNGPDGTI 1997û. )GPGVKE HCEVQTU VJCV FGVGTOKPG VJG RGTHQTOCPEG QH VJG EQPXGPVKQPCN [GCUV Saccharomyces cerevisiae KP DKQVGEJPQNQIKECN RTQEGUUGU CTG XGT[ YGNN KPXGUVKICVGF ýUGG %JCRVGTU 3-ôû. 6JG IGPGVKE DCUKU QH VJG MKNNGT RJGPQOGPQP KU YGNN WPFGTUVQQF KP S. cerevisiae CPF Kluyveromyces lactis ý5EJCHHTCVJ CPF $TGWPKI ùøøø; /CTSWKPC GV CN. ùøøù; 5EJOKVV CPF $TGKPKI ùøøùÿ UGG CNUQ %JCRVGTU 4 CPF ÷û. /QUV QH VJG DKQEQPVTQN [GCUVUÿ JQYGXGTÿ DGNQPI VQ VJG UQ-ECNNGF PQP-EQPXGPVKQPCN [GCUVUÿ YJGTG
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
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IGPGVKE U[UVGOU CTG PQV CU YGNN KPXGUVKICVGF CU HQT S. cerevisiae ýúGDGT CPF $CTVJ 19üü; 5RGPEGT GV CN. ùøøùû. 5Q HCT VJG IGPGVKE EJCTCEVGTKUVKEU QH DKQEQPVTQN [GCUVU JCXG OCKPN[ DGGP WUGF HQT VJG KFGPVKHKECVKQP CPF RJ[NQIGPGVKEU QH URGEKGU CPF UVTCKPUÿ WUKPI OQNGEWNCT VGEJPKSWGU ý/ELCWIJNKP GV CN. 199ø; öWTV\OCP 1994; öWTV\OCP CPF óQDPGVV 199ü; $GIGTQY GV CN. ùøøøû. *QYGXGTÿ VJG WUG QH IGPGVKE OGVJQFU HQT OCPKRWNCVKQP CPF CPCN[UKU QH VJG DKQEQPVTQN CEVKXKV[ JCU TCTGN[ DGGP FQEWOGPVGF ý,CPKUKGYKE\ CPF öQTUVGP ùøøùû. 6JQUG OGVJQFU YQWNF GPCDNG UGNGEVKQP QH DKQEQPVTQN OWVCPVUÿ IGPG VTCPUHGT WUKPI UGZWCN QT RCTCUGZWCN J[DTKFKUCVKQP CPF VTCPUHQTOCVKQPÿ GRKUVCVKE CPCN[UKUÿ OWNVK-EQR[ UWRRTGUUKQP CPF QXGTGZRTGUUKQP. 6JG EQORNGZKV[ QH VJG DKQEQPVTQN U[UVGOU OCMGU KV FKHHKEWNV VQ GUVCDNKUJ CRRTQRTKCVG UGNGEVKQP U[UVGOU HQT DKQEQPVTQN OWVCPVU. 6JG RQQT MPQYNGFIG CDQWV VJG IGPGVKE U[UVGOU QH VJG PQPEQPXGPVKQPCN DKQEQPVTQN [GCUVU RTGUGPVU CPQVJGT DCTTKGT. +P VJKU EJCRVGTÿ YG YKNN UWOOCTKUG EWTTGPV MPQYNGFIG CDQWV VJG IGPGVKEU QH RTQOKPGPV DKQEQPVTQN [GCUVU. 6JG CRRNKECVKQP QH ENCUUKECN CPF OQNGEWNCT IGPGVKE OGVJQFU KP VJG UVWF[ QH DKQEQPVTQN [GCUVU YKNN UWDUVCPVKCNN[ EQPVTKDWVG VQ VJG FGXGNQROGPV QH VJKU RTQOKUKPI HKGNF QH DKQVGEJPQNQI[ KP VJG HWVWTG.
10.2 #PVKHWPICl [GCUVU CPF VJGKT OQFGU QH KPJKDKVKQP 6JG CRRNKECVKQP CTGCU HQT DKQEQPVTQN [GCUVU KP CITKEWNVWTG CTG VJG EQPVTQN QH HWPICN RNCPV RCVJQIGPU KP ITGGPJQWUGU CPF HKGNFUÿ RQUVJCTXGUV FKUGCUGU ý2CWNKV\ CPF $oNCPIGT ùøø1; ,CPKUKGYKE\ CPF öQTUVGP ùøøùûÿ CPF QH OQWNFU FWTKPI UVQTCIG QH OQKUV EGTGCN ITCKP HQT CPKOCN HGGF ý2GVGTUUQP CPF 5EJP·TGT 199ô; &TWXGHQTU GV CN. ùøøùû. #VVGORVU VQ EQPVTQN YQQF RCVJQIGPU D[ [GCUVU JCXG DGGP TGRQTVGF ýúCNMGT GV CN. 199ô; 2C[PG GV CN. ùøøø; 2C[PG CPF $TWEG ùøø1û. 6JG OGEJCPKUOU QH KPJKDKVKQP YGTG OCKPN[ KPXGUVKICVGF D[ OKETQUEQR[ÿ RJ[UKQNQIKECNÿ CPF DKQEJGOKECN OGVJQFU. 6JQUG OGVJQFU CTG FKHHKEWNV VQ RGTHQTO KP VJG ÂTGCN• U[UVGO CPF HTGSWGPVN[ VJG UVCVGF OQFGU QH CEVKQP CTG OGTGN[ UWIIGUVKQPU VJCV CTG PQV GZRGTKOGPVCNN[ RTQXGP. %QORGVKVKQP HQT PWVTKGPVUÿ C OGEJCPKUO VJCV JCU HTGSWGPVN[ DGGP UWIIGUVGF VQ DG VJG KPJKDKVQT[ OGEJCPKUO ýUGG DGNQYû YCU KPXGUVKICVGF D[ CFFKPI INWEQUG QT QVJGT GUUGPVKCN EQORQWPFU VQ VJG U[UVGO ý&TQD[ GV CN. 19ü9; ©NCF GV CN. 1994û. òPG OGVJQF FGOQPUVTCVGF EQORGVKVKQP HQT PWVTKGPVU KP C DKQCUUC[ WUKPI RJ[UKECNN[ UGRCTCVGF CPVCIQPKUV CPF VCTIGV HWPIWU ý,CPKUKGYKE\ GV CN. ùøøøû. 10.2.1 ÿJ[llQURJGTG [GCUVU 2QYFGT[ OKNFGY HWPIK CTG KORQTVCPV ITGGPJQWUG RCVJQIGPU. 6JG DCUKFKQO[EGVQWU [GCUV-NKMG HWPIWU Ampelomyces quisqualis YCU UJQYP VQ CPVCIQPK\G RQYFGT[ OKNFGY URGEKGU KP VJG QTFGTU ©T[UKRJCNGUÿ /WEQTCNGUÿ CPF 2GTKURQTKCNGU QP EWEWODGTÿ TQUGÿ YJGCVÿ TGF ENQXGTÿ CPF CRRNG ýþCTYQQF 193ù; òFKPVUQXC 197ô; 2CWNKV\ CPF $oNCPIGT ùøø1û. #P QXGTXKGY QH DCUKFKQO[EGVQWU CPVKHWPICN [GCUVUÿ VCTIGV RCVJQIGPUÿ CPF VJG RTQVGEVGF U[UVGOU KU IKXGP KP 6CDNG 1. A. quisqualis KU C O[EQRCTC-
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8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
UKVG UJQYP VQ EQNQPK\G J[RJCG CPF EQPKFKQRJQTGU QH VJG RCVJQIGP; KPXCFKPI J[RJCG RCUUGF VJTQWIJ VJG EGNN YCNN QH VJG JQUV FKTGEVN[ KPVQ KVU RTQVQRNCUOÿ KVU URQTGU YGTG UJQYP VQ IGTOKPCVG KPVQ VJG J[RJCGÿ YJKEJ KP VWTP NGCF VQ VJG EQNNCRUG QH VJG RCVJQIGP ý©OOQPU 193ø; òFKPVUQXC 197ôû. A. quisqualis FGITCFGU EQNNQKFCN EJKVKP CPF NCOKPCTKP CPF C EQTTGNCVKQP DGVYGGP NCOKPCTKP FGITCFCVKQP CPF O[EQRCTCUKVKUO QP EWEWODGT RQYFGT[ OKNFGY ýSphaerotheca fuligineaû YCU FGOQPUVTCVGF ý*KjYGIGP 199ùDû. 6JG IGPWU Tilletiopsis EQPVCKPU QVJGT O[EQRCTCUKVGU VJCV ECP EQPVTQN RQYFGT[ OKNFGYÿ D[ EQNQPK\KPI J[RJCG CPF EQPKFKQRJQTGU QH VJG RCVJQIGP. +P EQPVTCUV VQ A. quisqualisÿ VJG[ FQ PQV RGPGVTCVG VJG J[RJCG ý*QEJ CPF 2TQXXKFGPVK 1979û. 6JGKT CPVKHWPICN CEVKXKV[ FGRGPFU QP UGETGVGF EQORQWPFU. ©ZVTCEGNNWNCT !-1.3-INWECPCUGÿ EJKVKPCUG CEVKXKV[ÿ CPF VJG CDKNKV[ VQ FGITCFG NCOKPCTKP JCXG DGGP FGOQPUVTCVGF. /QTGQXGTÿ C UGETGVGF HCVV[ CEKF GUVGT YKVJ CPVKHWPICN CEVKXKV[ YCU HQWPF ý*KjYGIGP 199ùD; *KjYGIGP 199ùC; ñTSWJCTV GV CN. 1994; ñTSWJCTV CPF 2WPjC ùøøùû. 6JG [GCUV Pseudozyma flocculosa ýQTKIKPCNN[ KFGPVKHKGF CU Sporothrix flocculosaÿ VGNGQOQTRJ Stephanoascus flocculosaû KPJKDKVU RQYFGT[ OKNFGY CPF QVJGT RCVJQIGPU D[ CPVKDKQUKU TCVJGT VJCP RCTCUKVKUO. 6JKU CPVKDKQUKU KU CEJKGXGF D[ VJG UGETGVKQP QH EGTVCKP HCVV[ CEKFU CEVKXG CICKPUV HWPIK YKVJ C JKIJ FGITGG QH WPUCVWTCVKQP QH RJQURJQNKRKF HCVV[ CEKFU CPF C NQY UVGTQN EQPVGPV KP VJGKT OGODTCPG ý$GP[CIQWD GV CN. 199÷C; $GP[CIQWD GV CN. 199÷D; #XKU CPF $oNCPIGT ùøøùû. #UEQO[EGVQWU CPF DCUKFKQO[EGVQWU [GCUVU QH VJG PCVWTCN UCRTQRJ[VKE RJ[NNQURJGTG HNQTC ýAureobasidium pullulansÿ Sporobolomyces roseusÿ Cryptococcus URR.ÿ Cladosporium URR.û GZGTV DKQEQPVTQN CEVKXKV[ QP XCTKQWU RNCPV RCVJQIGPUÿ RTQDCDN[ D[ UECXGPIKPI GZVTCPGQWU PWVTKGPVU ý$NCMGOCP CPF (QMMGOC 19üùû. (WTVJGT DCUKFKQO[EGVQWU CPF CUEQO[EGVQWU DKQEQPVTQN [GCUVU QH VJG RJ[NNQURJGTG CTG NKUVGF KP VJG 6CDNGU 1 CPF ù. 10.2.2 $KQEQPVTQl QH RQUVJCTXGUV FKUGCUGU 8CTKQWU OGEJCPKUOU CTG KORQTVCPV KP DKQEQPVTQN QH RQUVJCTXGUV FKUGCUGU. (TGSWGPVN[ C EQODKPCVKQP QH UGXGTCN OGEJCPKUOU KU HQWPF ý,CPKUKGYKE\ CPF öQTUVGP ùøøùû. 6JG [GCUV C. oleophilaÿ HQTOWNCVGF CU VJG EQOOGTEKCN DKQEQPVTQN CIGPV #URKTG½ÿ YCU FGOQPUVTCVGF VQ GZGTV CPVCIQPKUVKE CEVKXKV[ CICKPUV XCTKQWU OQWNFU QP C PWODGT QH FKHHGTGPV HTWKVU ýUGG 5EJGPC GV CN. ùøøø; ,CPKUKGYKE\ CPF öQTUVGP ùøøù; öCTCDWNWV GV CN. ùøøù CPF TGHGTGPEGU VJGTGKPû. 6JG UWIIGUVGF OGEJCPKUOU QH KPJKDKVKQP YGTG EQORGVKVKQP HQT URCEG CPF PWVTKGPVU CPF KPFWEVKQP QH JQUV TGUKUVCPEGÿ K.G. CEEWOWNCVKQP QH EJKVKPCUG CPF !-1.3-INWECPCUGÿ KPETGCUGF RTQFWEVKQP QH GVJ[NGPGÿ RJGP[NCNCPKPG COOQPKC N[CUG CPF RJ[VQCNGZKPGU ý2QTCV GV CN. ùøø1; &TQD[ GV CN. ùøøù; 2QTCV GV CN. ùøøùû. 6JG DCUKFKQO[EGVQWU [GCUV Cryptococcus albidus KU WUGF KP VJG EQOOGTEKCN DKQEQPVTQN RTQFWEV þKGNF 2NWU½. Crypt. albidus EQWNF CEV CU C DKQRTQVGEVKQP CIGPV CICKPUV B. cinerea QP EWEWODGTÿ VQOCVQÿ CPF DGCPU KP ITGGPJQWUG VTKCNU ý&KM CPF ©NCF 1999; &KM GV CN. 1999û CPF UVTCYDGTT[ KP HKGNF VTKCNU ý*GNDKI ùøøùû. /QTGQXGTÿ KV YCU FGOQPUVTCVGF VQ CEV CU C DKQEQPUGTXCVKQP CIGPV QP HTWKVU KP VJG RQUVJCTXGUV VTGCVOGPV CICKPUV B. cinerea CPF Pen. expansum QP CRRNG CPF RGCT ý(CP CPF 6KCP ùøø1; 6KCP GV CN. ùøøùDû. 6JKU [GCUV UGGOU OCKPN[ VQ CEV D[ EQORGVKVKQP HQT PWVTK-
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
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6CDNG 1. #PVKHWPICN DCUKFKQO[EGVQWU [GCUVU, [GCUV-NKMG HWPIK CPF TGNCVGF HWPIK KORGTHGEVK YKVJ DKQEQPVTQN CEVKXKV[ #PVKHWPICN [GCUV
+PJKDKVGF HWPIK
2TQVGEVGF U[UVGOU
Ampelomyces quisqualis ýU[P. Cicinnobolus cesatiiû
2QYFGT[ OKNFGY HWPIK ýErysiphe polygoni, E. cichoracearumÿ Sphaerotheca fuliginea CPF QVJGT HWPIK QH VJG QTFGTU Erysiphalesÿ Mucorales CPF Perisporialesû Botrytis cinerea, Penicillium expansum
%WEWODGTÿ TQUGÿ YJGCVÿ TGF ENQXGTÿ CRRNG
B. cinereaÿ Mucor periformisÿ Monilinia fructicolaÿ Pen. expansumÿ Phialophora malorumÿ Pezicula malicorticis Aspergillus niger, B. cinerea, Cochliobolus sativus, Penicillium digitatum, Pen. expansum, Pen. italicum, P. malorum, P. malicorticis, M. fructicola, Rhizopus stolonifer, Septoria nodorum
#RRNGÿ RGCTÿ UYGGV EJGTT[
%QORGVKVKQP HQT PWVTKGPVU
úJGCV NGCXGUÿ CRRNGÿ RGCTÿ UVTCYDGTT[ÿ MKYK HTWKVÿ VCDNG ITCRGÿ EKVTWU HTWKVUÿ UYGGV EJGTT[
%QORGVKVKQP HQT PWVTKGPVUÿ TGFWEVKQP QH RCVJQIGP UVKOWNCVKQP D[ VJG JQUVÿ EGNN YCNN FGITCFKPI GP\[OGU
2QYFGT[ OKNFGY HWPIK QH EWEWODGTÿ TQUG CPF YJGCVÿ B. cinerea, Cladosporium cucumerinum Rhodosporidium B. cinerea toruloides ýCPCOQTRJ Rhodotorula rubescensÿ U[P. Rhodotorula gracilisû
%WEWODGTÿ TQUGÿ YJGCVÿ DKQCUUC[
óGNGCUG QH VQZKE OGVCDQNKVGU ýCPVKOKETQDKCN HCVV[ CEKFU CPF VJGKT CNFGJ[FGUû %QORGVKVKQP HQT PWVTKGPVU
Cryptococcus albidus
Crypt. infirmominiatus
Crypt. laurentii
Pseudozyma flocculosa
5WIIGUVGF KPJKDKVKQP OGEJCPKUOU /[EQRCTCUKVKUOÿ FGITCFCVKQP QH EGNN YCNNU
%WEWODGTÿ %QORGVKVKQP HQT VQOCVQÿ PWVTKGPVU DGCPÿ UVTCYDGTT[ÿ CRRNGÿ RGCT
#RRNGÿ VQOCVQÿ DGCP ýRJ[NNQRNCPGûÿ IGTCPKWO NGCXGU
4GHGTGPEGU ý©OOQPU 193ø; *KjYGIGP 199ùD; 2CWNKV\ CPF $oNCPIGT ùøø1û ý©NCF GV CN. 1994; &KM GV CN. 1999; *GNDKI ùøøù; 6KCP GV CN. ùøøùDû ý5RQVVU GV CN. ùøøùû CPF TGHGTGPEGU VJGTGKP ý(QMMGOC 197ü; %JCPF)Q[CN CPF 5RQVVU 199÷D; %JCPF-)Q[CN CPF 5RQVVU 199÷C; %CUVQTKC GV CN. 1997; LKOC GV CN. 199ü; (KNQPQY 1999; LKOC GV CN. 1999; (KNQPQY ùøø1û ý#XKU CPF $oNCPIGT ùøøùû CPF TGHGTGPEGU VJGTGKP ý©NCF GV CN. 1994; (KNQPQY GV CN. 199÷; $WEM ùøøùû
3øù
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
6CDNG 1. ýEQPVKPWGFû Rhodotorula glutinis
A. niger, B. cinerea, P. digitatum, Pen. expansum, Pen. italicum, P. malorum, P. malicorticis
Sporobolomyces Alternaria porri, B. roseus cinerea, C. sativus, Colletotrichum graminicola, Pen. expansum, S. nodorum
Tilletiopsis spp.
2QYFGT[ OKNFGY HWPIK QH EWEWODGTÿ DCTNG[ CPF TQUG ýS. fuliginea, Erysiphe graminis, S. pannosa var. rosaeû
#RRNGÿ VCDNG ITCRGU UVTCYDGTT[ HTWKVU CPF NGCXGUÿ MKYK HTWKVÿ EKVTWU HTWKVUÿ RGCT úJGCVÿ OCK\Gÿ T[G CPF QPKQP NGCXGUÿ CRRNGÿ RGCT
%QORGVKVKQP HQT PWVTKGPVUÿ UGETGVKQP QH C UKFGTQRJQTGÿ KPFWEVKQP QH JQUV TGUKUVCPEG TGCEVKQPU
ý%JCPF-)Q[CN CPF 5RQVVU 199÷C; %CNXGPVG GV CN. 1999; LKOC GV CN. 1999; *GNDKI ùøø1û
%QORGVKVKQP HQT PWVTKGPVUÿ TGFWEVKQP QH RCVJQIGP UVKOWNCVKQP D[ VJG JQUV
ý(QMMGOC 197ü; (QMMGOC CPF FG 0QQKj 19ü1; ,CPKUKGYKE\ CPF $QTU 199ô; (KNQPQY ùøø1û CPF TGHGTGPEGU VJGTGKP ý*KjYGIGP 199ùD; öPWFUGP CPF 5MQW 1993; ñTSWJCTV GV CN. 1994û
%WEWODGTÿ /[EQRCTCUKVKUOÿ DCTNG[ÿ TQUG RTQFWEVKQP QH EGNN YCNN FGITCFKPI GP\[OGU
GPVUÿ CU UWIIGUVGF HTQO DKQEQPVTQN GZRGTKOGPVU CICKPUV B. cinereaÿ Pen. expansumÿ CPF Penicillium glabrumÿ DWV NQECNN[ KPFWEGF JQUV TGUKUVCPEG OC[ CNUQ RNC[ C TQNG ý©NCF GV CN. 1994; ©NCF GV CN. 199÷; 5KPKICINKC GV CN. 199ü; 6KCP GV CN. ùøøùCû. òVJGT URGEKGU QH VJG IGPWU Cryptococcus JCXG DGGP FGOQPUVTCVGF VQ EQPVTQN RTGJCTXGUV CPF RQUVJCTXGUV RCVJQIGPGU ý6CDNG 1û. +P CNN ECUGUÿ EQORGVKVKQP HQT PWVTKGPVU YCU UWIIGUVGF VQ DG VJG OGEJCPKUO QH DKQEQPVTQN. #PQVJGT OGEJCPKUO KU VJG EQPUWORVKQP QH DWV[N CEGVCVGÿ YJKEJ KU HQTOGF D[ CRRNGU CPF UVKOWNCVGU B. cinerea KPHGEVKQPU KP CRRNG YQWPFU. 5KOKNCT TGUWNVU EQPEGTPKPI TGFWEVKQP QH VJG DWV[N CEGVCVG EQPEGPVTCVKQP CPF KPJKDKVKQP YGTG QDVCKPGF YKVJ Sp. roseus ý(KNQPQY 1999; (KNQPQY ùøø1û. 5GETGVKQP QH !-1.3-INWECPCUG OKIJV CNUQ RNC[ C TQNG KP VJG KPJKDKVKQP QH B. cinerea CPF Pen. expansum D[ Crypt. laurentii ý%CUVQTKC GV CN. 1997û. 5GXGTCN QVJGT [GCUVU JCXG DGGP UJQYP VQ JCXG C RQVGPVKCN KP DKQEQPVTQN ý6CDNGU 1 CPF ùû. A. pullulansÿ Rhodosporidium toruloidesÿ CPF Rhodotorula glutinis YGTG CDNG VQ KPJKDKV RCVJQIGPU QP C XCTKGV[ QH RNCPVU CPF HTWKVU KP RTGJCTXGUV CPF RQUVJCTXGUV CRRNKECVKQPU. #ICKPÿ VJG OCjQT OGEJCPKUO YCU UWIIGUVGF VQ DG EQORGVKVKQP HQT PWVTKGPVUÿ DWV CNUQ VJG KPFWEVKQP QH JQUV TGURQPUGU OKIJV RNC[ C TQNG ýLKOC GV CN. 1999; +RRQNKVQ CPF 0KITQ ùøøø; *GNDKI ùøø1û. 6JG JKIJGUV FGITGG QH DKQEQPVTQN D[ R. toruloides CICKPUV B. cinerea QP IGTCPKWO NGCXGU YCU CEJKGXGF CV NQY INWEQUG EQPEGPVTCVKQPU ý$WEM ùøøùû.
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
3ø3
6CDNG ÿ. #PVKHWPICN CUEQO[EGVQWU [GCUVU, [GCUV-NKMG HWPIK CPF TGNCVGF HWPIK KORGTHGEVK YKVJ DKQEQPVTQN CEVKXKV[ #PVKHWPICN [GCUV
+PJKDKVGF HWPIK
Aureobasidium Alternaria porri, Aspullulans pergillus niger, Botrytis cinerea, Cochliobolus sativus, Drechslera sorokiniana, Penicillium digitatum, Pen. expansum, Pen. italcum, Pezicula malicorticis, Rhizopus stolonifer, Septoria nodorum Candida sp. B. cinerea, Pen. expansum, Monilinia fructicola Candida oleo- A. niger, B. cinerea, phila Geotrichum candidum, P. digitatum, Pen. expansum, Pen. italicum, M. fructicola, R. stolonifer Candida saitoana
Candida sake
B. cinerea, P. digitatum
B. cinerea, Pen. expansum, R. stolonifer, R. nigricans Debaryomyces A. niger, B. cinerea, hansenii ýCPC- Cladosporium herbaOQTRJ C. rum, Ophium piceae, famataû O. piliferum, Penicillium brevicompactum, Pen. expansum, P. digitatum, Pen. italicum, M. fructicola, R. stolonifer
2TQVGEVGF U[UVGOU úJGCVÿ T[G CPF QPKQP NGCXGUÿ EWEWODGTÿ VQOCVQÿ UVTCYDGTT[ÿ CRRNGÿ MKYK HTWKVÿ EKVTWU HTWKVUÿ VCDNG ITCRG
5WIIGUVGF KPJKDKVKQP OGEJCPKUOU %QORGVKVKQP HQT PWVTKGPVUÿ RTQFWEVKQP QH EGNN YCNN FGITCFKPI GP\[OGU
4GHGTGPEGU ý(QMMGOC 197ü; LGKDKPIGT GV CN. 1997; LKOC GV CN. 1999; 5EJGPC GV CN. 1999; ,CPKUKGYKE\ GV CN. ùøøø; %CUVQTKC GV CN. ùøø1û
5YGGV EJGTT[ÿ RGCEJÿ PGEVCTKPG %KVTWU HTWKVUÿ VCDNG ITCRGÿ EJGTT[ VQOCVQÿ RGCTÿ PGEVCTKPGU
ýöCTCDWNWV GV CN. ùøø1; öCTCDWNWV GV CN. ùøøùû %QORGVKVKQP ý5EJGPC GV CN. HQT URCEGðPW- ùøøø; &TQD[ GV VTKGPVUÿ KPFWE- CN. ùøøù; öCTCDWVKQP QH JQUV NWV GV CN. ùøøùû TGUKUVCPEG TGURQPUGU
#RRNGÿ EKVTWU HTWKVU
%QORGVKVKQP HQT PWVTKGPVUÿ RTQFWEVKQP QH EGNN YCNN FGITCFKPI GP\[OGUÿ KPFWEVKQP QH JQUV TGUKUVCPEG %QORGVKVKQP HQT PWVTKGPVU
#RRNGÿ RGCT #RRNGÿ VCDNG ITCRGUÿ UVTCYDGTT[ÿ MKYK HTWKVÿ EKVTWU HTWKVUÿ EJGTT[ VQOCVQÿ RGCEJÿ VQOCVQÿ UYGGV RQVCVQÿ UCRYQQF ý2KPWU UKNXGVTKUû
ý©N-)JCQWVJ GV CN. 199ü; ©N)JCQWVJ GV CN. ùøø1û
ý0WPGU GV CN. ùøøùû CPF TGHGTGPEGU VJGTGKP %QORGVKVKQP ý#TTCU 199÷; 5VGHQT URCEGð PW- XGPU GV CN. 1997; VTKGPVUÿ KPFWE- LKOC GV CN. 1999; VKQP QH JQUV 5EJGPC GV CN. TGUKUVCPEG TG- ùøøø; 2C[PG CPF URQPUGUÿ EQNQ- $TWEG ùøø1û PKUCVKQP CPF N[UKU QH HWPICN J[RJCG
3ø4
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
6CDNG ÿ. ýEQPVKPWGFû Metschnikowia pulcherrima ýCPCOQTRJ C. pulcherrimaû
B. cinerea, Pen. expansum
#RRNGÿ UVTCYDGTT[
%QORGVKVKQP HQT PWVTKGPVUÿ FKHHWUKDNG VQZKE OGVCDQNKVGU #RRNGÿ 2TQFWEVKQP QH ITCRGXKPG EGNN YCNN FGRNCPVUÿ ITCKPÿ ITCFKPI GPOQFGN CPK\[OGUÿ RTQOCNUÿ RNCVG FWEVKQP QH DKQCUUC[ MKNNGT VQZKPUÿ WPMPQYP OGEJCPKUOU
ý)WKPGDTGVKGTG GV CN. ùøøø; ,CPKUKGYKE\ GV CN. ùøø1; 5RCFCTQ GV CN. ùøøùû Pichia anomala Aspergillus caný5CYCPV GV CN. ýU[P. Hansenula didus, B. cinerea, 19ü9; úCNMGT GV anomalaÿ CPCPenicillium roqueCN. 199ô; %CUOQTRJ C. pellicufortii, P. carneum, UQPG GV CN. 1997; losaû P. paneum, P. ver/CINKCPK GV CN. rucosum, further 1997C; ,KjCMNK plant pathogenic CPF LGRQKXTG CPF YQQF FGEC[KPI 199ü; 2GVGTUUQP HWPIKÿ JWOCP RCVJQCPF 5EJP·TGT IGPKE HWPIK 1999; /CUKJ GV CN. ùøøøû P. guilliermondii Aspergillus flavus, #RRNGÿ %QORGVKVKQP ý/ELCWIJNKP GV ýCPCOQTRJ C. A. niger, B. cinerea, RGCEJÿ PGE- HQT URCEG CPF CN. 199ù; 2CUVGT guilliermondiiû G. candidum, Pen. VCTKPGÿ YKPG PWVTKGPVUÿ GV CN. 1993; #Tdigitatum, Pen. CPF VCDNG RTQFWEVKQP QH TCU GV CN. 199ü; italicum, R. stoloITCRGUÿ EKVEGNN YCNN FG- 5EJGPC GV CN. nifer TWU HTWKVUÿ ITCFKPI GPùøøø;
6JG [GCUV Pichia guilliermondii GZGTVU DKQEQPVTQN CEVKXKV[ QP C ITGCV PWODGT QH RQUVJCTXGUV RCVJQIGPU QP HTWKVU CPF ITCKPÿ DWV CNUQ KP C RTGJCTXGUV CRRNKECVKQP QP UVTCYDGTTKGU ý5EJGPC GV CN. ùøøø; )WGVUM[ GV CN. ùøøùû. $GUKFGU EQORGVKVKQP HQT URCEG CPF PWVTKGPVUÿ CVVCEJOGPV VQ J[RJCGÿ RTQFWEVKQP QH !-1ÿ3-INWECPCUG CPF UG-
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
3øô
ETGVKQP QH PQP-XQNCVKNG KPJKDKVQT[ UWDUVCPEGU YGTG QDUGTXGF VQ KPJKDKV RCVJQIGPKE HWPIK ýúKUPKGYUMK GV CN. 1991; #TTCU GV CN. 199ü; )WGVUM[ GV CN. ùøøùû. Pichia anomala YCU FGOQPUVTCVGF VQ GZGTV CPVKHWPICN CEVKXKV[ YKVJ C XCTKGV[ QH HWPIK KP JCDKVCVU CU FKXGTUG CU UVQTGF CRRNGUÿ ITCRGYKPG RNCPVUÿ CPF UVQTGF EGTGCN ITCKP. 5GXGTCN OGEJCPKUOU CTG UWIIGUVGF VQ KPHNWGPEG KVU RGTHQTOCPEG CU DKQEQPVTQN CIGPVÿ KPENWFKPI VJG RTQFWEVKQP QH UGETGVGF EGNN YCNN N[VKE GP\[OGU CPF MKNNGT VQZKPU ý,KjCMNK CPF LGRQKXTG 199ü; /CUKJ GV CN. ùøøø; &TWXGHQTU GV CN. ùøøù; (TGFNWPF GV CN. ùøøùû. +VU RGTHQTOCPEG OKIJV CNUQ DG EQPPGEVGF VQ KVU CDKNKV[ VQ ITQY CPF UWTXKXG KP OCP[ FKHHGTGPV CPF UVTGUUHWN GPXKTQPOGPVU. +V KU C PQPSaccharomyces YKPG [GCUV CPF JCU DGGP HQWPF KP [QIJWTV CPF QVJGT HQQF CPF KP UKNCIG HGTOGPVCVKQP ýG.I. öKVCOQVQ GV CN. 1999; %CIIKC GV CN. ùøø1; óQjCU GV CN. ùøø1û. 10.2.3 NQP-EQPXGPVKQPCl MKllGT [GCUVU KP DKQEQPVTQl 6JG RTQFWEVKQP QH MKNNGT VQZKPU D[ [GCUVU JCU RTQOKUG KP XCTKQWU DKQVGEJPQNQIKECN CRRNKECVKQPU QH CPVKHWPICN CEVKXKV[. öKNNGT [GCUVU UGETGVG RTQVGKPU VJCV CTG NGVJCN VQ UWUEGRVKDNG [GCUV UVTCKPU; CEVKXKV[ VQ HKNCOGPVQWU HWPIK CPF GXGP DCEVGTKC JCU CNUQ DGGP TGRQTVGF ýúCNMGT GV CN. 199ô; /CINKCPK GV CN. ùøøùû. 6JG MKNNGT UVTCKPU CTG TGUKUVCPV VQ VJGKT QYP RTQVGKP DWV OC[ DG UGPUKVKXG VQ MKNNGT RTQVGKPU QH QVJGT [GCUVU ý/CTSWKPC GV CN. ùøøù; 5EJOKVV CPF $TGKPKI ùøøùû. öKNNGT [GCUVU YKVJ RQVGPVKCN HQT DKQEQPVTQN CTG UWOOCTKUGF KP 6CDNG 3. Saccharomyces CPF PQP-Saccharomyces MKNNGT [GCUVU ECP CEV CU URQKNCIG [GCUVU KP DGGTÿ YKPGÿ CPF UCMo RTQFWEVKQP. 6JG[ ECP ECWUG UNQY CPF KPEQORNGVG HGTOGPVCVKQPUÿ YJKEJ EJCPIGU VJG EJCTCEVGTKUVKEU CPF VJG VCUVG QH VJG RTQFWEV ý$KUUQP 1999û. #VVGORVU YGTG OCFG VQ RTGXGPV UWEJ URQKNCIG D[ EQPUVTWEVKQP QH RTQFWEVKQP UVTCKPU RQUUGUUKPI VJG MKNNGT RJGPQV[RG ýG.I. 5GMK GV CN. 19üô; $QTVQN GV CN. 19ü÷û. /QTGQXGTÿ URQKNCIG QH UCNVGF HQQF NKMG OKUQÿ UQ[ UCWEGÿ QT UCNVGF XGIGVCDNGU D[ QUOQVQNGTCPV [GCUVU NKMG Zygosaccharomyces rouxii YCU HTGSWGPVN[ QDUGTXGF ý(NGGV 199ùû. *CNQVQNGTCPV MKNNGT [GCUVU CDNG VQ GNKOKPCVG VJQUG EQPVCOKPCPVU JCXG DGGP KUQNCVGF CPF EJCTCEVGTKUGF. 6JG WUG QH MKNNGT [GCUVU VQ EQODCV RNCPV CPF JWOCP RCVJQIGPUÿ YQQF FGEC[KPI HWPIKÿ CPF EQPVCOKPCPVU QH HQQF QT HGGF RTQFWEVKQP JCU DGGP EQPUKFGTGF ýUGG 6CDNG 3û. 6JG VJGTCRGWVKE RQVGPVKCN QH MKNNGT VQZKPU CPF QH CPVK-KFKQV[RKE CPVKDQFKGU VJCV CTG CDNG VQ OKOKE VJG CEVKQP QH MKNNGT VQZKPU CICKPUV HWPICN JWOCP RCVJQIGPU JCU DGGP FKUEWUUGF ý2QNQPGNNK GV CN. 19ü÷; /CINKCPK GV CN. 1997Cû. #PVK-KFKQV[RKE CPVKDQFKGU TGHNGEVKPI C P. anomala VQZKP JCXG DGGP FGOQPUVTCVGF VQ GZGTV C DTQCF CPVKOKETQDKCN URGEVTWO CPF YGTG UWEEGUUHWNN[ WUGF HQT VJG VTGCVOGPV QH C OQFGN ECPFKFCUKU KP TCV ý/CINKCPK GV CN. ùøøùû.
3ø÷
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
6CDNG 3. #PVKHWPICN RQVGPVKCN QH MKNNGT [GCUVU KP DKQEQPVTQN þKNNGT [GCUV Candida naeodendra Debaryomyces hansenii Pichia anomala Pichia farinosa Williopsis saturnus var. mrakii Williopsis saturnus var. saturnus Kluyveromyces drosophilarum K. marxianus Saccharomyces cerevisiae P. anomala W. saturnus var. mrakii W. saturnus var. saturnus P. anomala S. cerevisiae W. saturnus var. mrakii P. anomala S. cerevisiae W. saturnus var. mrakii K. lactis
2QVGPVKCN CRRNKECVKQP 2TGXGPVKQP QH URQKNCIG D[ QUOQVQNGTCPV [GCUVU KP UCNVGF HQQF ýG.I. OKUQÿ UQ[ UCWEGû
4GHGTGPEG ý#UJKFC GV CN. 19ü3; òJVC GV CN. 19ü4; 5W\WMK GV CN. 19ü9û
+PJKDKVKQP QH YQQF FGEC[KPI HWPIK
ýúCNMGT GV CN. 199ôû
+PJKDKVKQP QH RNCPV RCVJQIGPKE HWPIK
ýúCNMGT GV CN. 199ôû
+PJKDKVKQP QH JWOCP RCVJQIGPKE HWPIK
ýúCNMGT GV CN. 199ô; /CINKCPK GV CN. 1997Dû ýöKVCOQVQ GV CN. 1999û
2TGXGPVKQP QH URQKNCIG KP UKNQU
10.3 #XCKlCDlG KPHQTOCVKQP CDQWV IGPQOG QTICPKUCVKQP KP PQP-EQPXGPVKQPCl CPVKHWPICl [GCUVU 10.3.1 5GZWClKV[, IGPQOG UVTWEVWTG, CPF GZVTCEJTQOQUQOCl GlGOGPVU $CUKE MPQYNGFIG CDQWV VJG IGPQOG QTICPKUCVKQP KU PGEGUUCT[ HQT UWEEGUUHWN IGPGVKE OCPKRWNCVKQP. 6JKU KPHQTOCVKQP KPENWFGU RNQKF[ NGXGNÿ EJTQOQUQOG PWODGT CPF UK\Gÿ UGZWCNKV[ CPF OCVKPIÿ GZVTCEJTQOQUQOCN GNGOGPVUÿ CPF UGSWGPEG KPHQTOCVKQP ýUGG CNUQ %JCRVGT ùû. 10.3.1.1 Basidiomycetous yeasts and related imperfect species +P 19÷7ÿ C UGZWCN IGPGTCVKQP KP Rhodutorula glutinis YCU QDUGTXGF ý$CPPQ 19÷7û. 6JGUG UVTCKPU YGTG FGHKPGF CU VJG RGTHGEV UVCVG QH R. glutinisÿ Rhodosporidium toruloides. *QYGXGTÿ TGEGPVN[ÿ KV YCU FGOQPUVTCVGF QP VJG DCUKU QH VJG &1ð&ù UGSWGPEGU QH VJG NCTIG TKDQUQOCN UWDWPKV VJCV VJG V[RG UVTCKPU QH R. glutinis CPF R. toruloides CTG ENQUGN[ TGNCVGF DWV PQV EQPURGEKHKE ý(GNN GV CN. ùøøøû. *CRNQKF UVTCKPU QH R. toruloides CTG JGVGTQVJCNNKEÿ DGNQPIKPI VQ VJG OCVKPI V[RGU A QT aÿ JCXG C DWFFKPI OQTRJQNQI[ÿ CPF HQTO FKMCT[QVKE O[EGNKC CHVGT OCVKPI. &WTKPI URQTWNCVKQPÿ FKRNQKF VGNKQURQTGU CTG HQTOGF ý#DG GV CN. 197ôû. $CPPQ ý19÷7û UWIIGUVGF VJCV OGKQUKU RTQDCDN[ QEEWTU FWTKPI QWVITQYVJ QH VJGUG VGNKQURQTGU. *QYGXGTÿ CPCN[UKU
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
3ø7
QH VJG &0#-EQPVGPV CPF VJG UGITGICVKQP RCVVGTP QH CWZQVTQRJKE OCTMGTU UJQYGF VJCV PQ TGIWNCT OGKQUKU QEEWTTGF. 6JWUÿ OQUV UVTCKPU QH VJKU URGEKGU OKIJV DG CPGWRNQKF ý5COUQPQXC CPF ${VVEJGT 19üøû. 6JG UGZWCN E[ENG QH R. toruloides KU UWOOCTKUGF KP (KIWTG 1. /KVQVKECNN[ UVCDNG OQPQMCT[QVKE FKRNQKFU VJCV ITGY CU [GCUVU QT O[EGNKC EQWNF DG IGPGTCVGF D[ RTQVQRNCUV HWUKQP QH JCRNQKF UVTCKPU QH VJG UCOG CPF VJG QRRQUKVG OCVKPI V[RG ý5KRKE\MK CPF (GTGPE\[ 1977; $GEJGT CPF ${VVEJGT 19ü3û. 5GXGTCN IGPGU JCXG DGGP ENQPGF; OQUV QH VJGO CTG T&0# IGPGUÿ QVJGT IGPGU KPENWFG IGPGU GPEQFKPI HQT VJG RTGEWTUQT QH VJG OCVKPI V[RG RJGTQOQPG ý#MCFC GV CN. 19ü9û CPF VJG IGPG HQT VJG OGFKECNN[ CPF KPFWUVTKCNN[ KORQTVCPV GP\[OG RJGP[NCNCPKPG-COOQPKC-N[CUG ý2#Lû ý)KNDGTV GV CN. 19üôû. (GY HCEVU CTG MPQYP CDQWV VJG IGPGVKEU QH VJG [GCUV Crypt. albidus WUGF KP VJG EQOOGTEKCN CXCKNCDNG DKQEQPVTQN CIGPV þKGNF 2NWU½. )GPG UGSWGPEGU KP VJG ©/$LFCVCDCUG KPENWFG T&0# IGPGU CPF VJG !-1ÿ4-Z[NCPCUG GPEQFKPI IGPG ý$QWEJGT GV CN. 19üüû VJCV JCU DGGP WUGF CU OQFGN IGPG HQT JGVGTQNQIQWU IGPG GZRTGUUKQP KP UGXGTCN U[UVGOU ý/QTGCW GV CN. 199ù; /QTQUQNK GV CN. 1993; 2CUUQVJ CPF *CJP*kIGTFCN ùøøøû. A-HCEVQT A-EGNNU ýJCRNQKFû
a-EGNNU ýJCRNQKFû a-HCEVQT
2NQKF[TGFWEVKQP ýPQ TGIWNCT OGKQUKUû
(QTOCVKQP QH OCVKPI VWDGU
2NQKF[TGFWEVKQP ýPQ TGIWNCT OGKQUKUû
%GNN HWUKQPÿ HQTOCVKQP QH FKMCT[QVKE O[EGNKWO
(QTOCVKQP QH FKRNQKF VGNKQURQTGU
(KI. 1. 5GZWCN E[ENG QH Rhodosporidium toruloides CEEQTFKPI VQ ý$CPPQ 19÷7; #DG GV CN. 197ô; 5COUQPQXC CPF ${VVEJGT 19üøûÿ HQT GZRNCPCVKQP UGG VJG VGZV.
3øü
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
6JG IGPQOG QH VJG KORGTHGEV [GCUV Pseud. flocculosa YCU CPCN[UGF D[ %*©( GNGEVTQRJQTGUKU. #V NGCUV GNGXGP EJTQOQUQOGU KP VJG UK\G TCPIG QH ø.ôô- ù.9 /DR YGTG HQWPF ý%JGPI GV CN. ùøø1û. òPN[ TKDQUQOCN UGSWGPEGU CTG EWTTGPVN[ CXCKNCDNG KP VJG ©/$L-FCVCDCUG. 10.3.1.2 Asexual ascomycetous yeasts 6JG CUGZWCN CUEQO[EGVQWU [GCUV-NKMG HWPIWU A. pullulans ýDNCEM [GCUVû KU RQQTN[ KPXGUVKICVGF CV VJG IGPGVKE NGXGN. 5GXGTCN IGPG UGSWGPEGU CTG CXCKNCDNG KP VJG ©/$L-FCVCDCUGÿ OCKPN[ IGPGU QH TKDQUQOCN &0#. 6JG URA3 IGPG JCU DGGP ENQPGF CPF CPCN[UGF ýóQUG GV CN. ùøøøû. 6JG [GCUV C. oleophila WUGF CU VJG EQOOGTEKCN DKQEQPVTQN CIGPV #URKTG½ KU IGPGVKECNN[ RQQTN[ KPXGUVKICVGF. #RCTV HTQO T&0# IGPGUÿ QPN[ QPG RTQVGKP-GPEQFKPI IGPG KU EJCTCEVGTKUGF. 6JG GP\[OG GPEQFGF D[ VJKU IGPG KU C UGETGVGF 1ÿ3-!INWECPCUGÿ YJKEJ RTQDCDN[ RNC[U C TQNG KP VJG DKQEQPVTQN CEVKXKV[ QH VJKU [GCUV ý5GICN GV CN. ùøøùû. &KTGEV UGNGEVKQP QH ura3 QT leu2 OWVCPVU WUKPI VJG ô-(ò# ý$QGMG GV CN. 19ü4û QT VJG CNRJC COKPQ CFKRCVG OGVJQF ý%JCVVQQ GV CN. 1979ûÿ TGURGEVKXGN[ÿ YCU PQV UWEEGUUHWN. óCPFQO OWVCIGPGUKU YKVJ ©/5 ýùøï UWTXKXQTUû [KGNFGF 14 CWZQVTQRJKE OWVCPVU HTQO 3.÷‚1øô OWVCIGPKUGF EGNNU; VYGNXG QH VJGO TGSWKTGF JKUVKFKPG ý%JCPF-)Q[CN GV CN. 1999û. 10.3.1.3 Debaryomyces hansenii /QUV UVTCKPU QH VJG QUOQVQNGTCPV [GCUV D. hansenii VJCV UJQYGF DKQEQPVTQN CEVKXKV[ QP HTWKVU CPF YQQF ý5VGXGPU GV CN. 1997; 2C[PG CPF $TWEG ùøø1û CTG JCRNQKF. 6JG[ TCTGN[ OCVG CPF FKRNQKFK\G VTCPUKGPVN[ D[ UQOCVQICOQWU CWVQICO[. #UEK EQPVCKP QPG UKPING URQTG ýöTGIGT XCP óKj CPF 8GGPJWKU 197ô; XCP FGT úCNV GV CN. 1977û. LKPGCT RNCUOKFU YGTG HQWPF KP VJKU [GCUV VJCV UJQYGF UKOKNCTKV[ VQ VJG MKNNGT RNCUOKFU QH Kluyveromyces lactisÿ JQYGXGTÿ VJG[ YGTG PQV EQPPGEVGF VQ MKNNGT CEVKXKV[. 6JG RNCUOKFU YGTG UVCDKNK\GF D[ QUOQVKE RTGUUWTGÿ IGPGTCVGF D[ JKIJ EQPEGPVTCVKQPU QH UCNVÿ UQTDKVQNÿ QT IN[EGTQN ý)WPIG GV CN. 1993û. D. hansenii XCT. hansenii YCU TGEGPVN[ KPXGUVKICVGF KP VJG EQORCTCVKXG IGPQOKE Â)oPQNGXWTGU• RTQjGEV . # PWODGT QH IGPGU YGTG KFGPVKHKGFÿ KPENWFKPI VJG OKVQEJQPFTKCN IGPQOG CPF PWENGCT IGPGU. òXGTTGRTGUGPVCVKQP QH IGPGU KPXQNXGF KP VJG VTCPURQTV HCEKNKVCVKQP OKIJV RQKPV VQ VJG RJ[UKQNQIKECN RGEWNKCTKVKGU QH D. hansenii EQORCTGF VQ S. cerevisiae ýLoRKPING GV CN. ùøøø; 5QWEKGV GV CN. ùøøøû. 10.3.1.4 Pichia guilliermondii and Candida guilliermondii P. guilliermondii KU C JGVGTQVJCNNKE [GCUV CPF JCRNQKF UVTCKPU QH VJG OCVKPI V[RGU OCVî CPF OCV- CTG CDNG VQ HQTO UVCDNG FKRNQKF J[DTKFU ýúGDGT CPF $CTVJ 19üüû. $[ C OQFKHKECVKQP QH VJG QTKIKPCN J[DTKFKUCVKQP OGVJQF 11ü UVTCKPU HQTOGTN[ ENCUUKHKGF CU C. guilliermondii YGTG UGZWCNN[ ETQUUGF CPF TGENCUUKHKGF CU P. guilliermondii ý<JCTQXC GV CN. 1977û. $[ TGRGCVGF E[ENGU QH ETQUUKPI CPF UGNGEVKQP QH VJG DGUV URQTWNCVKPI J[DTKFU C IGPGVKE NKPG YCU QDVCKPGF ECRCDNG QH RTQFWEKPI C NCTIG PWODGT QH URQTGU ý5KDKTP[ GV CN. 1977û. 5KZ NKPMCIG ITQWRU JCXG DGGP HQWPF UQ HCT D[
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
3ø9
IGPGVKE CPCN[UKU QH CWZQVTQRJKE OWVCPVU ý2TCJN 19üüû. óGEGPVN[ VJG PWODGT QH EJTQOQUQOGU QH VJG V[RG UVTCKPU QH P. guilliermondii CPF C. guilliermondii YCU FGOQPUVTCVGF VQ DG CV NGCUV GKIJV ý$CK GV CN. ùøøøû. # EQPUKFGTCDNG XCTKCDKNKV[ QH VJG EJTQOQUQOCN DCPFKPI RCVVGTP YCU HQWPF KP QVJGT UVTCKPU QH VJG JGVGTQIGPQWU CPCOQTRJ C. guilliermondii. òPN[ HGY IGPG UGSWGPEGU CTG CXCKNCDNG KP VJG ©/$L FCVCDCUGÿ K.G. VJQUG IGPGU QH TKDQUQOCN EQORQPGPVU CPF IGPGU KPXQNXGF KP VJG TKDQHNCXKP U[PVJGUKU. 10.3.1.5 Pichia anomala 6JG IGPGVKEU QH VJG [GCUV P. anomala JCXG TGEGPVN[ DGGP KPXGUVKICVGF ý0CWOQX GV CN. ùøø1û. 6JG PCVWTCN RNQKF[ NGXGN YCU UWIIGUVGF VQ DG FKRNQKF. *GVGTQVJCNNKE VGUVGT UVTCKPU GZKUV DWV COQPI KUQNCVGU HTQO FKHHGTGPV UQWTEGUÿ UVCDNG OCVKPI V[RGU YGTG UGNFQO QDUGTXGF. 5GZWCN J[DTKFKUCVKQP YCU CEJKGXGF DGVYGGP UGXGTCN UVTCKPUÿ DWV VJG RTQFWEVU GZJKDKVGF RQQT HGTVKNKV[ CPF IGPGTCVGF PQP-XKCDNG CUEQURQTGU. 6JG OCjQTKV[ QH J[DTKFU YCU RQN[RNQKFÿ RTQDCDN[ VGVTCRNQKFÿ UQOG UVTCKPU HQTOGF FKRNQKF J[DTKFU YKVJ TGIWNCT OGKQVKE UGITGICVKQP. 5KOKNCT VQ OCP[ QVJGT [GCUV URGEKGU ý5EJGTGT CPF /CIGG 199ø; 2CUUQVJ GV CN. 199ù; $CK GV CN. ùøøøûÿ C TGOCTMCDNG EJTQOQUQOG NGPIVJ RQN[OQTRJKUO YCU FGVGEVGF. *QYGXGTÿ CNN UVTCKPU DGNQPIGF VQ VJG UCOG URGEKGU CEEQTFKPI VQ VJG UGSWGPEGU QH VJGKT &1ð&ù TGIKQPU ý0CWOQX GV CN. ùøø1û. 6JG UGSWGPEGU QH UGXGTCN IGPGU CTG CXCKNCDNG KP VJG ©/$L-FCVCDCUGÿ KPENWFKPI C IGPG GPEQFKPI CP GZVTCEGNNWNCT GZQ-!-1ÿ3-INWECPCUG YKVJ CPVKOKETQDKCN CEVKXKV[. 10.3.1.6 Mitochondrial genetics 6JG [GCUVU C. guilliermondiiÿ Kloeckera apiculataÿ P. anomalaÿ W. mrakiiÿ CPF W. saturnus YGTG UJQYP VQ DG RGVKVG PGICVKXGÿ K.G. VJG[ FQ PQV VQNGTCVG OWVCVKQPU KP VJG OKVQEJQPFTKCN IGPQOG ý$WNFGT 19÷4û. W. mrakii YCU VJG HKTUV [GCUV VJCV YCU FGOQPUVTCVGF VQ JCXG C NKPGCT OKVQEJQPFTKCN IGPQOG. +VU UK\G YCU FGVGTOKPGF VQ DG ôô MD ýúGUQNQYUMK CPF (WMWJCTC 19ü1û. /QUV QH VJG OKVQEJQPFTKCN IGPQOGU CTG TGICTFGF VQ DG EKTEWNCTÿ JQYGXGTÿ VJG QEEWTTGPEG QH NKPGCT OKVQEJQPFTKCN &0# KP QVJGT URGEKGU JCU DGGP FGOQPUVTCVGF ý(WMWJCTC GV CN. 1993; ,CEQDU GV CN. 199÷û. 10.3.2 )GPGVKEU QH VJG MKllGT EJCTCEVGT KP PQP-EQPXGPVKQPCl DKQEQPVTQl [GCUVU 6JG HKTUV IGPGVKE KPXGUVKICVKQPU QP VJG MKNNGT EJCTCEVGT KP S. cerevisiae CPF K. lactis FGOQPUVTCVGF VJCV VJKU RJGPQOGPQP YCU WPFGT EQPVTQN QH E[VQRNCUOKE FGVGTOKPCPVU VJCV TGSWKTGF VJG RTGUGPEG QH FQOKPCPV PWENGCT IGPGU ý5QOGTU CPF $GXCP 19÷9; 6KRRGT CPF $QUVKCP 19ü4û. *QYGXGTÿ VJG IGPGVKE DCUKU QH VJG MKNNGT RJGPQOGPQP ECP DG SWKVG XCTKCDNG. +P S. cerevisiaeÿ FUó0# XKTWUGU JCXG DGGP KFGPVKHKGF VQ DG VJG QTKIKP QH VJG MKNNGT RJGPQV[RG ý5EJOKVV CPF $TGKPKI ùøøùû. +P K. lactisÿ VJG MKNNGT IGPGU CTG NQECNK\GF QP NKPGCT FU&0# RNCUOKFU ý5EJCHHTCVJ CPF $TGWPKI ùøøøÿ UGG CNUQ %JCRVGT ÷û. FUó0# XKTWUGU CPF NKPGCT FU&0# RNCUOKFU
31ø
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
YGTG FGOQPUVTCVGF VQ GPEQFG VJG MKNNGT EJCTCEVGT KP QVJGT [GCUVUÿ DWV CNUQ EJTQOQUQOCN KPJGTKVCPEG JCU DGGP HQWPF ý/CINKCPK GV CN. 1997D; /CTSWKPC GV CN. ùøøù; 5EJOKVV CPF $TGKPKI ùøøùû. # PWENGCT IGPG GPEQFKPI VJG UCNV OGFKCVGF MKNNGT VQZKP ý5/ö VQZKPû QH VJG JCNQVQNGTCPV URGEKGU P. farinosa JCU DGGP ENQPGF CPF EJCTCEVGTK\GF. 6JG VQZKP UJQYGF KVU OCZKOWO CEVKXKV[ CV ù M 0C%N. 6JG PWENGQVKFG UGSWGPEG RTGFKEVGF C RTGRTQVQZKP YKVJ ùùù COKPQ CEKF TGUKFWGUÿ EQPVCKPKPI C UKIPCN UGSWGPEG CPF VYQ UWDWPKVU QH VJG VQZKP. /CVWTKPI YCU UWIIGUVGF VQ QEEWT D[ RTQEGUUKPI YKVJ VJG ö©:ù-NKMG GPFQRGRVKFCUG CPF VJG ö©:1-NKMG ECTDQZ[RGRVKFCUG CPF VJG OQNGEWNCT YGKIJV QH VJG OCVWTG $F FKOGT YCU GUVKOCVGF VQ 14 M&C ý5W\WMK CPF 0KMMWPK 1994û. öKNNGT CEVKXKV[ JCU DGGP FGOQPUVTCVGF KP OCP[ FKHHGTGPV KUQNCVGU QH VJG [GCUV P. anomala ýG.I. þCR GV CN. ùøøø; (TGFNWPF GV CN. ùøøùû. %QPUKFGTCDNG FKHHGTGPEGU KP VJG CEVKXKV[ URGEVTWO QT VJG OQNGEWNCT EJCTCEVGTKUVKEU QH VJG VQZKPU YGTG QDUGTXGF DGVYGGP FKHHGTGPV UVTCKPU ýöCIK[COC GV CN. 19üü; þCR GV CN. ùøøøû. /QNGEWNCT OCUUGU QH 43 M&Cÿ ü3-üô M&Cÿ CPF üô M&C YGTG FGVGTOKPGF HQT FKHHGTGPV KUQNCVGF MKNNGT RTQVGKPU ý5CYCPV GV CN. 19ü9; 8WUVKP GV CN. 19ü9; )W[CTF GV CN. 1999û. 8GT[ NCTIG MKNNGT VQZKPU ýCDQWV 3øø M&Cû YGTG KUQNCVGF CPF EJCTCEVGTKUGF HTQO VYQ JCNQVQNGTCPV P. anomala UVTCKPU ýöCIK[COC GV CN. 19üüû. #NN VJG KUQNCVGF P. anomala MKNNGT RTQVGKPU YGTG EQPUKFGTCDN[ FKHHGTGPV TGICTFKPI COKPQ CEKF EQORQUKVKQPÿ MKNNGT URGEVTWOÿ QRVKOWO R* CPF VGORGTCVWTGÿ FGITGG QH IN[EQU[NCVKQP CPF KUQGNGEVTKE RQKPV. +P QPG ECUGÿ VJG VQZKP YCU GZVTGOGN[ VJGTOQ UVCDKNG; MKNNGT CEVKXKV[ YCU UVKNN QDUGTXGF CHVGT KPEWDCVKQP CV ÷øº% HQT HKXG JQWTU CPF FKUCRRGCTGF QPN[ CHVGT ùô OKP CV 1øøº% ý8WUVKP GV CN. 19ü9û. $QVJ MKNNGT VQZKPU HTQO VJG JCNQVQNGTCPV P. anomala UVTCKPU YGTG JGCV NCDKNG VQ FKHHGTGPV FGITGGU ýöCIK[COC GV CN. 19üüû. #RRNKECVKQP QH E[ENQJGZKOKFGÿ GNGXCVGF VGORGTCVWTGUÿ GVJKFKWO DTQOKFGÿ QT CETKFKPG QTCPIG FKF PQV EWTG VJG MKNNGT RJGPQV[RGÿ KPFKECVKPI EJTQOQUQOCN KPJGTKVCPEG ýþQWPI CPF þCIKW 197ü; öCIK[COC GV CN. 19üü; 8WUVKP GV CN. 19ü9û. # UKOKNCT XCTKCDKNKV[ QH VJG MKNNGT EJCTCEVGTKUVKEU JCU CNUQ DGGP QDUGTXGF KP QVJGT [GCUVUÿ KPENWFKPI W. saturnus XCT. mrakiiÿ K. lactisÿ CPF P. membranefaciens ý#UJKFC GV CN. 19ü3; þCOCOQVQ GV CN. 19ü÷; þCR GV CN. ùøøøû. +P W. saturnus XCT. mrakii CPF KP W. saturnus XCT. saturnus PWENGCT MKNNGT VQZKP IGPGU JCXG DGGP ENQPGF ýHMK CPF HSKÿ TGURGEVKXGN[û. $QVJ IGPGU JCF C JKIJ FGITGG QH KFGPVKV[. 6JG[ GPEQFGF RTGEWTUQTU VQ MKNNGT VQZKPU QH 1ùô CPF 1ù4 COKPQ CEKFUÿ TGURGEVKXGN[ÿ EQPVCKPKPI C UKIPCN RGRVKFG CPF RTQRGRVKFG YJKEJ KU RTQDCDN[ RTQEGUUGF D[ C ö©:ù-NKMG RTQVGCUG. $QVJ IGPGU EQPHGTTGF VJG MKNNGT EJCTCEVGT VQ S. cerevisiae YJGP VTCPUHGTTGF KPVQ VJKU [GCUV ýöKOWTC GV CN. 1993û. òVJGT IGPGU YKVJ JQOQNQI[ VQ VJG HSK IGPG OC[ GZKUV KP W. saturnus XCT. mrakii ýöKOWTC GV CN. 199ôDû.
10.4 #XCKlCDlG IGPGVKE OGVJQFU HQT DKQEQPVTQl [GCUVU òPG QH VJG OQUV KORQTVCPV IGPGVKE CRRTQCEJGU VQ DKQEQPVTQN KU VJG KFGPVKHKECVKQP CPF RJ[NQIGPGVKE ENCUUKHKECVKQP QH [GCUV URGEKGU CPF UVTCKPU D[ OQNGEWNCT OGVJQFU ý#XKU GV CN. ùøø1Dû. óGNKCDNGÿ HCUVÿ CPF UKORNG OGVJQFU HQT UVTCKP CPF URGEKGU KFGP-
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
311
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ý)WNNKPQ GV CN. 199ô; #XKU GV CN. ùøø1Dû. 10.4.1 /QlGEWlCT OGVJQFU HQT KFGPVKHKECVKQP CPF RJ[lQIGPGVKE CPCl[UKU QH DKQEQPVTQl [GCUVU /QNGEWNCT OGVJQFU CTG PQY CXCKNCDNG VJCV CNNQY VJG KFGPVKHKECVKQP QH [GCUVU HCUVGT CPF HTGSWGPVN[ OQTG TGNKCDNG VJCP VJG ENCUUKECN RJ[UKQNQIKECN VGUVU. 6CDNG 4 RTQXKFGU C UWTXG[ CDQWV CXCKNCDNG OQNGEWNCT OGVJQFU HQT UVTCKP CPF URGEKGU KFGPVKHKECVKQP. 6JG TGIKQP GPEQFKPI VJG TKDQUQOCN ó0# ýT&0#û KU RTQOKUKPI HQT RJ[NQIGPGVKE UVWFKGU CPF URGEKGU KFGPVKHKECVKQP DGECWUG KV EQPVCKPU DQVJ EQPUGTXGF CPF XCTKCDNG TGIKQPUÿ CNNQYKPI VJG FGVGTOKPCVKQP QH FKUVCPV CPF ENQUG TGNCVKQPUJKRU ýöWTV\OCP CPF óQDPGVV 199ü; 8CNGPVG GV CN. 1999û. 5GSWGPEKPI VJG URGEKGUURGEKHKE &1ð&ù TGIKQP QH VJG NCTIG UWDWPKV T&0# CPF VJG KPVGTPCN VTCPUETKDGF URCEGT ý+65û TGIKQP KU EWTTGPVN[ VJG UVCPFCTF VGEJPKSWG VQ KFGPVKH[ [GCUVU CV VJG URGEKGU NGXGN ý(KI. ùÿ öWTV\OCP 1994; öWTV\OCP CPF óQDPGVV 199ü; $CK GV CN. ùøøùû. óGUVTKEVKQP HTCIOGPV NGPIVJ RQN[OQTRJKUO ýó(L2û QH VJG +65 RTQXKFGU C TCRKF OGVJQF HQT VJG KFGPVKHKECVKQP QH [GCUV URGEKGU ý©UVGXG-
31ù
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
5GSWGPEG CPCN[UKU
2TKOGTí0L1
2TKOGTí0L4 &1ð&ù-TGIKQPÿ ÷øø DR
1ü5 T&0#
ô.ü5 T&0#
+65
ùü5 T&0#
+65
ì4øø
üøø DR
2TKOGTUí +65ôÿ +651
2TKOGTí +654
1ü5 T&0#
ùô5 T&0#
ó(L2ÿ UGSWGPEG CPCN[UKU
(KI. ÿ. 5EJGOCVKE FTCYKPI QH VJG T&0# TGIKQP QH [GCUVU. 2TGHGTTGF TGIKQPU HQT URGEKGU KFGPVKHKECVKQP CTG VJG KPVGTPCN VTCPUETKDGF URCEGTU ý+65û CPF VJG ô•-GPF QH VJG NCTIG TKDQUQOCN UWDWPKV ý&1ð&ù-TGIKQPÿ PWENGQVKFGU ÷3-÷4ù QH VJG RDN25 IGPG QH S. cerevisiaeû. 6JG &0# KU CORNKHKGF WUKPI RTKOGTU ý+65ôÿ +651ÿ +654; 0L1ÿ 0L4û VJCV DKPF VQ EQPUGTXGF UVTGVEJGU QH VJG T&0# CPF HWTVJGT CPCN[UGF. 2TKOGT UGSWGPEGU CTG IKXGP KP 8CNGPVG GV CN. ý1999û. 6JG HKIWTG KU FTCYP CEEQTFKPI VQ VJG OCR QH S. cerevisiae EJTQOQUQOG :++ CU IKXGP KP VJG Saccharomyces IGPQOG FCVCDCUG ý5)&ÿ JVVRíððIGPQOG-YYY.UVCPHQTF.GFWð5CEEJCTQO[EGUðû.
HCEG QH HTWKVU CPF XGIGVCDNGUÿ CNN QH VJGO YGTG CDNG VQ EQPVTQN UGXGTCN RCVJQIGPKE HWPIK ý5EJGPC GV CN. 1999û. # UKOKNCT FKXGTUKV[ YCU HQWPF COQPI KUQNCVGU HTQO QVJGT GPXKTQPOGPVUÿ KPENWFKPI TQEMUÿ NGCXGUÿ CPF HTWKV ýñT\t GV CN. 1999û. 6JG UGSWGPEG QH VJG 1ü5 Tó0# YCU WUGF VQ FGUKIP C URGEKGU-URGEKHKE RTQDG HQT A. pullulans ýLK GV CN. 199÷û. 6JKU RTQDG YCU CRRNKGF HQT HNWQTGUEGPEG KP UKVW J[DTKFKUCVKQP ýHKUJû VQ KFGPVKH[ VJG [GCUV QP UNKFGU CPF NGCXGU CPF VQ OQPKVQT VJG FGXGNQROGPV QH
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
313
6CDNG ý. /QNGEWNCT OGVJQFU HQT [GCUV KFGPVKHKECVKQP /GVJQF 5GSWGPEKPI TGIKQPU QH T&0# ý&1ð&ùÿ +65û óGUVTKEVKQP HTCIOGPV NGPIVJ RQN[OQTRJKUO ýó(L2û QH VJG +65 2WNUGF HKGNF IGN GNGEVTQRJQTGUKU ý2()©û
+FGPVKHKECVKQP NGXGN 5RGEKGU
óGHGTGPEG ý8CNGPVG GV CN. 1999û
5RGEKGU
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5VTCKPÿ URGEKGUÿ XCTKGV[
G.I. ý8CWIJCP-/CTVKPK GV CN. ùøøø; 0CWOQX GV CN. ùøø1û ýúKNNKCOU GV CN. 199ø; *CFT[U GV CN. 199ùû ýúGNUJ CPF /E%NGNNCPF 199øû G.I. ý/ELCWIJNKP GV CN. 199ø; 8CWIJCP-/CTVKPK GV CN. ùøøøû
óCPFQO CORNKHKGF RQN[- 5VTCKP OQTRJKE &0# ýó#2&û #TDKVTCT[ RTKOGF 2%ó 5VTCKP ý#2-2%óû &0#-&0# TGCUUQEKCVKQP 5VTCKPÿ URGEKGUÿ XCTKGV[
GV
CN.
VJG A. pullulans RQRWNCVKQP KP VJG CRRNG RJ[NNQRNCPG KP C HKGNF GZRGTKOGPV ýLK GV CN. 1997; 5RGCT GV CN. 1999; #PFTGYU GV CN. ùøøùû +P C NCTIG VCZQPQOKE UVWF[ÿ ôøø CUEQO[EGVQWU [GCUVU YGTG CPCN[UGF D[ UGSWGPEKPI VJG &1ð&ù TGIKQPU. 6JG FKXGTIGPEG KP VJKU FQOCKP YCU CNYC[U UWHHKEKGPV VQ FKUVKPIWKUJ KPFKXKFWCN URGEKGU. %QPURGEKHKE UVTCKPU JCF NGUU VJCP 1ï PWENGQVKFG UWDUVKVWVKQP KP VJKU TGIKQP. 2J[NQIGPGVKE VTGGU IGPGTCVGF QP VJG DCUKU QH VJG &1ð&ù TGIKQP CPF VJG UGSWGPEG QH VJG 1ü5 T&0# YGTG EQORCTGF VQ GCEJ QVJGT CPF TGUWNVGF KP C JKIJ FGITGG QH EQPEQTFCPEG. 6JWUÿ EQORCTKPI VJG UGSWGPEGU QH VJG &1ð&ù TGIKQPU QH VJG NCTIG TKDQUQOCN UWDWPKV RTQXKFGU C TGNKCDNG RQYGTHWN VQQN HQT KFGPVKHKECVKQP CPF RJ[NQIGPGVKE KPXGUVKICVKQPU. $KQEQPVTQN [GCUVU YGTG FKUVTKDWVGF QXGT UGXGTCN ENCFGU KP VJKU CPCN[UKUÿ HQT KPUVCPEG P. anomala CPF W. saturnus DGNQPIGF VQ VJG P. anomala ENCFG VJCV KPENWFG UGXGTCN OGODGTU QH VJG HQTOGT IGPWU Hansenula; C. oleophilaÿ D. hanseniiÿ CPF P. farinosa YGTG ITQWRGF KP VJG Debaryomycesð Lodderomyces ENCFG CPF M. pulcherima DGNQPIGF VQ VJG Stephanoascusð Metschnikowa ENCFG ýöWTV\OCP CPF óQDPGVV 199üû. &TQD[ GV CN. ý1999û WUGF ó#2&ÿ #2-2%óÿ +65 UGSWGPEKPI CPF RJ[UKQNQIKECN EJCTCEVGTKUCVKQP VQ EJCTCEVGTKUG VJG GRKRJ[VKE [GCUV RQRWNCVKQP QP ITCRGHTWKV. +P VJKU GZRGTKOGPVÿ JQYGXGTÿ ó#2& CPF #2-2%ó FKF PQV RTQXKFG UCVKUHCEVQT[ TGUQNWVKQP QH VJG [GCUV UVTCKPU. òPN[ VYQ FKHHGTGPV RCVVGTPU YGTG HQWPF COQPI KUQNCVGU YKVJ JKIJ DKQEQPVTQN CEVKXKV[ÿ CNVJQWIJ VJG PWODGT QH URGEKGU YCU CV NGCUV UKZ ýC. guilliermondiiÿ C. sakeÿ C. oleophilaÿ D. hanseniiÿ Crypt. albidusÿ Crypt. laurentiiû CU GUVKOCVGF D[ RJ[UKQNQIKECN OGVJQFU CPF D[ VJG UGSWGPEG QH VJG +65. 6JGUG TGUWNVU UVTQPIN[ UWIIGUV VJCV KFGPVKHKECVKQP DCUGF QP TCPFQO CORNKHKECVKQP PGGFU VQ DG XGTKHKGF D[ CFFKVKQPCN OGVJQFU.
314
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
10.4.2 /GVJQFU HQT VJG OCPKRWlCVKQP QH DKQEQPVTQl [GCUVU #XCKNCDNG OGVJQFU HQT OCPKRWNCVKQP ENCUUKECNN[ KPENWFG OWVCIGPGUKUÿ UGZWCN QT RCTCUGZWCN J[DTKFKUCVKQPÿ PGY OGVJQFU KPENWFG VTCPUHQTOCVKQP CPF VJG KPVTQFWEVKQP QH JGVGTQNQIQWU IGPGU ýúGDGT CPF $CTVJ 19üü; úCPI GV CN. ùøø1; 5RGPEGT GV CN. ùøøùû. 6JGUG OGVJQFU ECP DG WUGF VQ KFGPVKH[ RCVJYC[U KPXQNXGF KP DKQEQPVTQN. (QT KPUVCPEGÿ UVTCKPU ECTT[KPI OWVCVKQPU VJCV ECWUG FGETGCUGF DKQEQPVTQN CEVKXKV[ ECP DG KUQNCVGF. 6JQUG OWVCVKQPU ECP DG EQODKPGF WUKPI VJG CDQXG-OGPVKQPGF OGVJQFU QH IGPG VTCPUHGT GPCDNKPI GRKUVCVKE CPCN[UKU CPF VJWUÿ KFGPVKHKECVKQP QH VJG PWODGT QH DKQEQPVTQN OGEJCPKUOU. /QTGQXGTÿ OWVCVKQPU CTG TGSWKTGF CU OCTMGT HQT UGZWCN CPF RCTCUGZWCN J[DTKFKUCVKQP CPF HQT VTCPUHQTOCVKQP. (TGSWGPVN[ÿ VJQUG OCTMGTU KPENWFG OWVCVKQPU VJCV ECWUG FGRGPFGPE[ QP EGTVCKP COKPQ CEKFUÿ XKVCOKPUÿ RWTKPGUÿ QT R[TKOKFKPGU. 6JG KPFWEVKQP QH VJQUG OWVCVKQPU ECP DG CEJKGXGF D[ EJGOKECN OWVCIGPGUKU QT ñ8- QT KQPKUKPI TCFKCVKQP. /WVCPVU ECP DG UETGGPGF D[ CWZCPQITCRJ[ÿ WUKPI FTQRQWV OGFKWO ýóQUG GV CN. 199øû. &KTGEV UGNGEVKQP OGVJQFU JCXG DGGP FGXGNQRGF HQT VJG KPFWEVKQP QH ura3 CPF leu2 OWVCPVU KP S. cerevisiae ý%JCVVQQ GV CN. 1979; $QGMG GV CN. 19ü4û. 6JGUG OGVJQFU CTG PQV IGPGTCNN[ CRRNKECDNG VQ PQP-EQPXGPVKQPCN [GCUVU CPF OWUV DG VGUVGF HQT GCEJ URGEKHKE URGEKGU ý%JCPF-)Q[CN GV CN. 1999û. 5GZWCN J[DTKFKUCVKQP OC[ DG RQUUKDNG KP C RGTHGEV URGEKGU. *QYGXGTÿ KUQNCVGU HTQO VJG GPXKTQPOGPV HTGSWGPVN[ JCXG C NQY HGTVKNKV[ QT HQTO OCKPN[ PQP-XKCDNG URQTGU ýG.I. 0CWOQX GV CN. ùøø1û. (QT IGPGVKE CPCN[UKU VJG DTGGFKPI QH C IGPGVKE NKPG YKVJ JKIJ URQTG HQTOCVKQP TCVG CPF C JKIJ RTQRQTVKQP QH XKCDNG URQTGU KU HTGSWGPVN[ PGEGUUCT[ ý5KDKTP[ GV CN. 1977û. 5GZWCN J[DTKFKUCVKQP ýOCVKPIû ECP DG CEJKGXGF D[ OKZKPI QH JCRNQKF UVTCKPU QH VJG QRRQUKVG OCVKPI V[RG ýKP VJG ECUG QH JGVGTQVJCNNKE URGEKGUû YKVJ GCEJ QVJGT. +P VJG ECUG QH JQOQVJCNNKE UVTCKPUÿ VYQ FKHHGTGPVN[ OCTMGF UVTCKPU ECP DG OKZGF CPF J[DTKFU ECP DG UGNGEVGFÿ DWV UGNHFKRNQKFKUCVKQP OC[ NGCF VQ EQPHWUKPI TGUWNVU ý/GNCMG GV CN. 199÷û. *GPEGÿ VGEJPKSWGU HQT UGZWCN J[DTKFKUCVKQP PGGF VQ DG QRVKOKUGF HQT GXGT[ URGEKGU ý<JCTQXC GV CN. 1977; /GNCMG GV CN. 199÷û. (QT KORGTHGEV URGEKGU QT KH CVVGORVU QH UGZWCN J[DTKFKUCVKQP HCKN KP C RGTHGEV URGEKGUÿ UVTCKPU ECP DG J[DTKFKUGF D[ RCTCUGZWCN OGCPUÿ WUKPI RTQVQRNCUV HWUKQP ýUGG CNUQ %JCRVGT ùû. 6JKU VGEJPKSWG GXGP CNNQYU KPVGTURGEKGU J[DTKFKUCVKQP ý5RGPEGT GV CN. 19üü; úGDGT CPF $CTVJ 19üü; öCXCPCIJ CPF úJKVVCMGT 199÷û. *QYGXGTÿ RTQDCDN[ DGECWUG QH VJG CTVKHKEKCN EGNN HWUKQPÿ WPWUWCN GXGPVU NKMG EJTQOQUQOCN TGCTTCPIGOGPVU JCXG DGGP QDUGTXGF ýöNKPPGT CPF ${VVEJGT 19üôû. òP VJG QVJGT JCPFÿ CPCN[UKU QH RTQVQRNCUV HWUKQP RTQFWEVU ECP NGCF VQ VJG FGXGNQROGPV QH CRRTQRTKCVG OGVJQFU HQT CEJKGXGOGPV QH UGZWCN J[DTKFKUCVKQP ý/GNCMG GV CN. 199÷û. 6TCPUHQTOCVKQP KU C XGT[ RQYGTHWN OGVJQF QH IGPG VTCPUHGT KP [GCUV. ñUKPI CRRTQRTKCVG XGEVQTUÿ QXGTGZRTGUUKQP QH IGPGUÿ VCTIGVGF QT TCPFQO OWVCIGPGUKU ECP DG CEJKGXGF ý)KGV\ CPF úQQFU ùøø1; úCPI GV CN. ùøø1û. 6JG OQUV HTGSWGPVN[ WUGF OGVJQFU VQ IGPGTCVG EQORGVGPV EGNNU CTG VJG VTGCVOGPVU YKVJ DKXCNGPV CNMCNKPG KQPUÿ GURGEKCNN[ LKùîÿ GNGEVTQRQTCVKQP CPF URJGTQRNCUV VTCPUHQTOCVKQP. $GECWUG VTCPUHQTOCVKQP KU C TCTG GXGPVÿ CRRTQRTKCVG FQOKPCPV OCTMGTU CTG TGSWKTGF VQ UGNGEV VTCPUHQTOCPVU VJCV EQORNGOGPV CP CWZQVTQRJ[ÿ EQPHGT CP CPVKDKQVKE TGUKUVCPEG QT RTQXKFG EQNQWTKPI QT HNWQTGUEGPEG ýúCPI GV CN. ùøø1û. 8GEVQTU HQT VTCPUHQTOCVKQP ECP
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
31ô
DG GRKUQOCN QT KPVGITCVKXG. (QT GRKUQOCN XGEVQTUÿ CP CWVQPQOQWUN[ TGRNKECVKPI UGSWGPEG ý#ó5û KU PGEGUUCT[ VJCV HWPEVKQPU KP VJG TGURGEVKXG JQUV. (TGSWGPVN[ÿ VJG #ó5 QH VJG S. cerevisiae ù±-RNCUOKF FQGU PQV HWPEVKQP KP PQPEQPXGPVKQPCN [GCUVUÿ CPF QVJGT #ó5-UGSWGPEGU VJWU PGGF VQ DG KUQNCVGF ýúCPI GV CN. ùøø1û. 6JG CDQXG-OGPVKQPGF OGVJQFU JCXG DGGP WUGF VQ OCPKRWNCVG [GCUVU YKVJ DKQEQPVTQN RQVGPVKCN. +P R. toruloidesÿ CWZQVTQRJKE OWVCPVU YGTG KUQNCVGF D[ TCPFQO OWVCIGPGUKU CPF GPTKEJGF D[ KPQUKVQN UVCTXCVKQP ý6WNN[ 19üôû. # VTCPUHQTOCVKQP U[UVGO JCU DGGP GUVCDNKUJGFÿ WUKPI VJG URJGTQRNCUV OGVJQF YKVJ VJG S. cerevisiae LEU2-IGPG QT VJG PAL IGPG GPEQFKPI RJGP[NCNCPKPG COOQPKC-N[CUG CU UGNGEVCDNG OCTMGT. 2CN- UVTCKPU CTG WPCDNG VQ WUG RJGP[NCNCPKPG CU ECTDQP QT PKVTQIGP UQWTEG. 6JG ù± QTKIKP FKF PQV HWPEVKQP KP R. toruloidesÿ VJG VTCPUHQTOCVKQPU [KGNFGF GKVJGT UVCDNG KPVGITCPVU QT PQP-UVCDNG GRKUQOCN VTCPUHQTOCPVU ý6WNN[ CPF )KNDGTV 19üôû. # OGVJQF HQT VJG VTCPUHQTOCVKQP QH Pseud. flocculosa YCU GUVCDNKUJGF ý%JGPI GV CN. ùøø1û. (QT A. pullulansÿ C VTCPUHQTOCVKQP U[UVGO JCU DGGP FGXGNQRGF WUKPI URJGTQRNCUVU CPF C IGPG EQPHGTTKPI TGUKUVCPEG VQ J[ITQO[EKP $ CU UGNGEVCDNG OCTMGT ý%WNNGP GV CN. 1991; 6JQTPGYGNN GV CN. 199ôû. #PQVJGT VTCPUHQTOCVKQP U[UVGO CRRNKGF VJG A. pullulans ura3 IGPG CU UGNGEVCDNG OCTMGT. 5VTCKPU YKVJ C FGHKEKGPE[ KP VJKU IGPG YGTG KUQNCVGF WUKPI EJGOKECN OWVCIGPGUKU CPF ô-(ò# UGNGEVKQP. 6JG IGPG YCU KUQNCVGF D[ J[DTKFKUCVKQP QH C RCTVKCN NKDTCT[ YKVJ C RTQDG HTQO C RTGXKQWUN[ KUQNCVGF IGPG HTCIOGPV ýóQUG GV CN. ùøøøû. (QT VJG [GCUV P. anomalaÿ C VTCPUHQTOCVKQP U[UVGO JCU DGGP FGXGNQRGF HQT C JCRNQKF UVTCKP FGTKXGF HTQO C FKRNQKF KUQNCVG HTQO YCUVGYCVGT VTGCVOGPV ýòICVC GV CN. 199ôû. #P ura3 OWVCVKQP YCU KPFWEGF KP VJKU UVTCKP D[ EJGOKECN OWVCIGPGUKU CPF UWDUGSWGPV ô-(ò# UGNGEVKQP. 6JKU OWVCPV YCU VTCPUHQTOGF YKVJ C EQPUVTWEV ECTT[KPI VJG JQOQNQIQWU URA3 IGPG QH P. anomala. ñTCî UVTCKPU ECTTKGF VJG RNCUOKF KPVGITCVGF KP VJG IGPQOG D[ JQOQNQIQWU TGEQODKPCVKQP ýòICVC GV CN. 199ôû. 6JG P. anomala URA3 IGPG YCU CNUQ UJQYP VQ EQORNGOGPV VJG JQOQNQIQWU ura3 OWVCPVU QH S. cerevisiae. +PVGTGUVKPIN[ÿ VJG UGSWGPEG FGUETKDGF D[ òICVC GV CN. ý199ùû FKHHGTGF D[ 1ø COKPQ CEKFU HTQO VJG UGSWGPEG QH C IGPG VJCV YCU KUQNCVGF HTQO CPQVJGT UVTCKP ý©/$L CEEGUUKQP PWODGT þø9ùù1û TGHNGEVKPI KPVTCURGEKHKE XCTKCPEG YKVJKP VJG URGEKGU P. anomala. 6JG DKQEQPVTQN CIGPV C. oleophila YCU VTCPUHQTOGF WUKPI GKVJGT VJG S. cerevisiae HIS5 IGPG VQ EQORNGOGPV C *KU- OWVCPV QT VJG E. coli hph IGPG VJCV EQPHGTTGF TGUKUVCPEG CICKPUV J[ITQO[EKP $ VQ VJG TGEKRKGPVU ý%JCPF-)Q[CN GV CN. 1999; þGJWFC GV CN. ùøø1û. 6JG RNCUOKFU KPVGITCVGF UVCDN[ KP VJG IGPQOG; KH VJG JQOQNQIQWU UGSWGPEG QH VJG C. oleophila EXG1 IGPG HNCPMGF VJG VTCPUHQTOKPI &0#ÿ EQTTGEV JQOQNQIQWU KPVGITCVKQP YCU QDUGTXGF KP UKZ QH GKIJV VTCPUHQTOCPVU. +H VJG XGEVQT FKF PQV EQPVCKP CP[ JQOQNQIQWU UGSWGPEGUÿ KV KPVGITCVGF TCPFQON[ KPVQ VJG IGPQOG. 6JWUÿ OGVJQFU HQT DQVJ TCPFQO CPF VCTIGVGF IGPG FKUTWRVKQP CTG PQY CXCKNCDNG KP VJKU [GCUVÿ RTQXKFKPI RQYGTHWN VQQNU HQT GZRNQTKPI VJG OGVCDQNKE DCUKU QH DKQEQPVTQN CPF HQT VJG NCDGNNKPI QH DKQEQPVTQN UVTCKPU GPCDNKPI VJGKT TGEQXGT[ ý%JCPF-)Q[CN GV CN. 1999; þGJWFC GV CN. ùøø1û.
31÷
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
10.þ 6JG WUG QH IGPGVKE OGVJQFU HQT VJG OCPKRWlCVKQP QH DKQEQPVTQl CEVKXKV[ 10.þ.1 #RRlKECVKQP QH OWVCPVU KP DKQEQPVTQl U[UVGOU 6JG WUG QH OWVCIGPGUKU VQ OCPKRWNCVG VJG DKQEQPVTQN CEVKXKV[ QH XCTKQWU PQPEQPXGPVKQPCN [GCUV UVTCKPU KU C RTQOKUKPI VQQN VQ QRVKOKUG VJG RGTHQTOCPEG QH VJG TGURGEVKXG DKQEQPVTQN CIGPV. #NVJQWIJ VJKU VGEJPKSWG JCU TCTGN[ DGGP WUGF ý,CPKUKGYKE\ CPF öQTUVGP ùøøùûÿ C DTQCF HKGNF HQT VJG CRRNKECVKQP QH OWVCIGPGUKU KU QDXKQWU. 6Q QDVCKP OWVCPVU VJCV CTG WUGHWN HQT DKQEQPVTQN VJG GUVCDNKUJOGPV QH CP CRRTQRTKCVG UGNGEVKQP U[UVGO KU VJG OQUV ETKVKECN UVGR. %JCTCEVGTKUVKEU VJCV CTG FKTGEVN[ KPXQNXGF KP DKQEQPVTQN CPF VJCV ECP DG EJCPIGF D[ OWVCIGPGUKU CTG HQT KPUVCPEG VJG CDKNKV[ VQ CVVCEJ VQ VJG JQUV UWTHCEGÿ VQ ITQY HCUVÿ VQ EQPUWOG PWVTKGPVU YKVJ C JKIJ TCVGÿ CPF VQ RTQFWEG KPJKDKVQT[ EQORQWPFU. /WVCVKQPU VJCV KORTQXG VJQUG EJCTCEVGTKUVKEU YKNN KORTQXG VJG DKQEQPVTQN RGTHQTOCPEG QH VJG YJQNG U[UVGOÿ YJGTGCU PWNN OWVCPVU YKNN JGNR VQ WPTCXGN VJG DKQEQPVTQN OGEJCPKUOU. $KQEQPVTQN CIGPVU OWUV HWNHKN UGXGTCN TGSWKTGOGPVUÿ G.I. TGUKUVCPEG CICKPUV GPXKTQPOGPVCN UVTGUUGU VJCV JGNRU VJG CPVCIQPKUV VQ UWTXKXG QP VJG JQUV ý+RRQNKVQ CPF 0KITQ ùøøøûÿ TGUKUVCPEG CICKPUV HWPIKEKFGUÿ RGUVKEKFGU QT CPVKOGVCDQNKVGU YJGP WUGF KP CP KPVGITCVKXG CRRTQCEJÿ PQ PGICVKXG KPVGTCEVKQPU YKVJ QVJGT DKQEQPVTQN QTICPKUOU YJGP WUGF KP EQODKPCVKQPÿ CPF PQ NQUU QH DKQEQPVTQN CEVKXKV[ YJGP WUGF KP EQOOGTEKCN HQTOWNCVKQPU ý2CWNKV\ CPF $oNCPIGT ùøø1; ,CPKUKGYKE\ CPF öQTUVGP ùøøùû. /WVCIGPGUKU RTQOKUGU VQ DG WUGHWN VQ KUQNCVG DKQEQPVTQN UVTCKPU YKVJ VJG FGUKTGF EJCTCEVGTKUVKEU. /WVCPVU YKVJ VJG CDQXG-OGPVKQPGF EJCTCEVGTKUVKEU OKIJV DG KUQNCVGF KP FKHHGTGPV YC[U. %GNNU ECP DG OWVCIGPKUGF WUKPI TCPFQO OWVCIGPGUKU CPF CRRTQRTKCVG OWVCPVU ECP DG UGNGEVGF HTQO CEEQTFKPIN[ FGUKIPGF KP XKVTQ CUUC[U ýóQUG GV CN. 199øû. 2QUKVKXG OWVCPVUÿ HQT KPUVCPEG YKVJ C TGUKUVCPEG CICKPUV C EGTVCKP HWPIKEKFG OC[ DG KUQNCVGF HTQO EWNVWTGU EQPUVCPVN[ VTGCVGF YKVJ VJG HWPIKEKFG ý*KjYGIGP 19ü÷; $GP[CIQWD CPF $oNCPIGT 199ôû. 6JG OKVQVKE UVCDKNKV[ QH KUQNCVGF OWVCVKQPU KU CP KORQTVCPV RCTCOGVGTÿ DQVJ HQT VJG RGTHQTOCPEG CPF HQT TGNKCDKNKV[ QH VJG DKQEQPVTQN U[UVGO CPF HQT UGEWTKV[ TGCUQPU. 6JG WUG QT KUQNCVKQP QH UGXGTCN OWVCPVU KP VJG DKQEQPVTQN GPXKTQPOGPV JCU DGGP FGUETKDGF. *KjYGIGP ý19ü÷û KUQNCVGF C UVTCKP QH Tilletiopsis minor VJCV EQWNF ITQY KP VJG RTGUGPEG QH 1øø ±IðON QH VJG HWPIKEKFG HGPCTKOQN. +P Pseud. flocculosa OWVCPVU TGUKUVCPV CICKPUV VJG U[UVGOKE HWPIKEKFG FQFGOQTRJ-CEGVCVG YGTG KUQNCVGF D[ TGRGCVGF GZRQUWTG VQ ITQYKPI COQWPVU QH VJG HWPIKEKFG CPF TGKUQNCVKQP QH ITQYKPI EGNNU. $[ VJKU CRRTQCEJÿ C OWVCPV VQNGTCVKPI WR VQ 3øø ±IðON EQWNF DG UGNGEVGFÿ C FQUCIG VJCV KU ôøï JKIJGT VJCP TGEQOOGPFGF HQT VJG VTGCVOGPV QH RQYFGT[ OKNFGY QP TQUGU. 6JG OWVCPVU FKF PQV FKHHGT HTQO VJG YKNF V[RG KP KVU DKQEQPVTQN CEVKXKV[. +P UVCDKNKV[ VGUVUÿ VJG HWPIKEKFG TGUKUVCPEG QH VJG OWVCPV FKF PQV FGETGCUG CHVGT 1ø VTCPUHGTU KP OGFKWO YKVJQWV VJG HWPIKEKFG ý$GP[CIQWD CPF $oNCPIGT 199ôû. (QT VJG UCOG URGEKGUÿ VJG KUQNCVKQP QH OWVCPVU WPCDNG VQ RTQFWEG CPVKHWPICN EQORQPGPVU JCU DGGP TGRQTVGF. 6JQUG OWVCPVU YKNN DG WUGHWN HQT VJG WPTCXGNNKPI QH VJG DKQU[PVJGUKU RCVJYC[ QH VJGUG CPVKDKQVKEU CPFÿ VJGTGHQTGÿ HQT VJG WPFGTUVCPFKPI QH VJG OGEJCPKUO QH DKQEQPVTQN RTQXKFGF D[ VJKU [GCUV ý2CWNKV\ CPF $oNCPIGT ùøø1û.
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
317
+P UQOG CRRTQCEJGUÿ DKQEQPVTQN CIGPVU YGTG EQODKPGF YKVJ PWVTKGPV CPCNQIWGU NKMG ù-FGUQZ[-&-INWEQUG ýù-&ò)û ý,CPKUKGYKE\ 1994; ,CPKUKGYKE\ CPF öQTUVGP ùøøùû. 6JQUG CPVKOGVCDQNKVGU ECP CNUQ KPJKDKV VJG DKQEQPVTQN CIGPVÿ CU FGOQPUVTCVGF HQT Sp. roseus ý,CPKUKGYKE\ 1994û. 6JKU [GCUV YCU WUGF KP C EQODKPGF CRRTQCEJ VQIGVJGT YKVJ Pseudomonas syringae CPF ù-&ò). +P EQPVTCUV VQ P. syringaeÿ Sp. roseus YCU UWUEGRVKDNG VQ 1ø OIðON QH VJG PWVTKGPV CPCNQIWG. 5RQPVCPGQWU OWVCPVU YGTG KUQNCVGF D[ NQPI VKOG EWNVKXCVKQP KP OGFKC EQPVCKPKPI ù-&ò). òPG UGNGEVGF OWVCPV YCU RTQXGP VQ DG UVCDNGÿ DGECWUG PQ NQUU QH TGUKUVCPEG QEEWTTGF CHVGT 1ø EQPUGEWVKXG VTCPUHGTU CV VYQ FC[U KPVGTXCNU QP PQP-UGNGEVKXG OGFKWO. 6JKU OWVCPVÿ EQODKPGF YKVJ ù-&ò) RTQXKFGF C ITGCVN[ GPJCPEGF DKQEQPVTQN CEVKXKV[ QH Pen. expansum QP CRRNGÿ YJGP EQORCTGF YKVJ VJG YKNF V[RG. 10.þ.2 #RRlKECVKQP QH IGPGVKECll[ GPIKPGGTGF UVTCKPU KP DKQEQPVTQl U[UVGOU 6JG KPVTQFWEVKQP QH HQTGKIP IGPGU KP C DKQEQPVTQN UVTCKP ECP DG WUGHWN KP UGXGTCN YC[U. # WPKSWG IGPG UGSWGPEG ECP DG VJG DCUKU QH FGVGEVKQP QH VJG UVTCKP VJWU CNNQYKPI KFGPVKHKECVKQP QH TG-KUQNCVGU HTQO VJG DKQEQPVTQN GPXKTQPOGPV. ©CU[ KFGPVKHKECVKQP CPF OQPKVQTKPI ECP DG QDVCKPGF KH C EGTVCKP EJCTCEVGTKUVKEÿ G.I. HNWQTGUEGPEG YKNN DG VTCPUHGTTGF VQ VJG DKQEQPVTQN UVTCKP. $GUKFGU VJG WUG HQT UVTCKP KFGPVKHKECVKQPÿ IGPGVKE GPIKPGGTKPI ECP CNUQ DG WUGF HQT VJG UVTCKP KORTQXGOGPV. #NN VJG KUUWGU QH UVTCKP KORTQXGOGPV VJCV ECP DG CRRTQCEJGF D[ OWVCIGPGUKU ECP CNUQ DG RGTHQTOGF WUKPI IGPGVKE GPIKPGGTKPIÿ KP OQUV ECUGU KP C OQTG FKTGEVGF YC[. 5GXGTCN GZCORNGU QH WUKPI IGPGVKECNN[ GPIKPGGTGF UVTCKPU KP DKQEQPVTQN JCXG DGGP FGUETKDGF. # )(2 XCTKCPV JCU DGGP GZRTGUUGF KP A. pullulans CPF QVJGT [GCUVU ý0KITQ GV CN. 1999; 8CPFGP ú[OGNGPDGTI GV CN. 1999û. 6JG VTCPUHQTOGF [GCUV EQWNF GCUKN[ DG OQPKVQTGF QP NGCH UWTHCEGUÿ VJWU GPCDNKPI DKQOQPKVQTKPI CPF SWCPVKHKECVKQP QH VJG DKQEQPVTQN CIGPV. +P CFFKVKQPÿ VJG TGRQTVGT IGPG ECP CNUQ DG WUGF VQ GZRNQTG TGIWNCVKQP QH IGPG GZRTGUUKQP KP A. pullulans ý8CPFGP ú[OGNGPDGTI GV CN. 1999û. #PQVJGT TGRQTVGT IGPGÿ GPEQFKPI VJG !-INWEWTQPKFCUG QH E. coli ý)ñ5û YCU KPVTQFWEGF KP C. oleophila ý%JCPF-)Q[CN GV CN. 199ü; %JCPF-)Q[CN GV CN. 1999û. #NVJQWIJ VJKU IGPG YCU PQV GZRTGUUGF KP VJG [GCUVÿ KV YCU CP GZEGNNGPV VCTIGV HQT UVTCKP KFGPVKHKECVKQP. 6TCPUHQTOCPVU YGTG PQV KPHNWGPEGF KP VJGKT RGTHQTOCPEG KP VJG DKQRTQVGEVKQP QH EKVTWU HTWKVU CICKPUV ITGGP OQWNF ýP. digitatumû CPF VJGKT RQRWNCVKQP F[PCOKEU YCU OQPKVQTGF D[ 2%ó. 6JG WUG QH MKNNGT VQZKPU KP DKQEQPVTQN JCU HTGSWGPVN[ DGGP FKUEWUUGF CPF UGXGTCN GZCORNGU QH VJGKT CRRNKECVKQP CTG PQY CXCKNCDNG ý/CTSWKPC GV CN. ùøøù; 5EJOKVV CPF $TGKPKI ùøøùû. # K. lactis UVTCKP JCU DGGP EQPUVTWEVGF VQ RTGXGPV CGTQDKE FGVGTKQTCVKQP KP UKNCIG. #GTQDKE FGVGTKQTCVKQP KU ECWUGF D[ [GCUVU VJCV EQPUWOG NCEVKE CEKF KP VJG UKNQU YJGP QZ[IGP DGEQOGU CXCKNCDNG CHVGT QRGPKPIÿ VJWU NGCFKPI VQ CP KPETGCUG QH VJG R* CPF NQUUGU QH FT[ OCVVGT CPF PWVTKGPVU. # K. lactis MKNNGT UVTCKP YCU FKUTWRVGF KP KVU PCK1 IGPGÿ GPEQFKPI VJG RJQURJQGPQNR[TWXCVG ECTDQZ[MKPCUG ý2©2%öû. 6JKU IGPG KU C MG[ IGPG HQT INWEQPGQIGPGUKU CPFÿ VJGTGHQTGÿ GUUGPVKCN HQT VJG ITQYVJ QP NCEVKE CEKF. 6JG TGUWNVKPI OWVCPV YCU WPCDNG VQ EQPUWOG NCEVKE CEKFÿ JCF VJG UCOG MKNNGT CEVKXKV[ CU VJG YKNFV[RGÿ CPF YCU UJQYP VQ KPJKDKV P. anomala
31ü
8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
KP C OQFGN UKNQ GZRGTKOGPV ýöKVCOQVQ GV CN. 199ü; öKVCOQVQ GV CN. 1999û. 6JG MKNNGT RTQVGKPU QH VJG IGPWU Williopsis OKIJV DG GURGEKCNN[ RTQOKUKPI HQT EQPVTQN QH URQKNCIG KP HGGF CPF HQQF GPXKTQPOGPVU DGECWUG QH VJGKT JKIJ UVCDKNKV[ QXGT C DTQCF R* CPF VGORGTCVWTG TCPIG CPF KVU DTQCF MKNNKPI URGEVTWO ý/CINKCPK GV CN. 1997Dû. W. mrakii MKNNGT VQZKP YCU JGVGTQNQIQWUN[ RTQFWEGF KP A. niger D[ GZRTGUUKQP QH C HWUKQP QH VJG */ö IGPG QH W. saturnus XCT. mrakii YKVJ VJG INWEQCO[NCUG IGPG QH A. niger. 6JG RTQFWEGF RTQVGKP GZGTVGF MKNNGT CEVKXKV[ CICKPUV [GCUVU KUQNCVGF HTQO HQQF CPF UKNCIG CPF YCU UVCDNG KP C YKFG TCPIG QH R* CPF VGORGTCVWTG. òPUGV QH CGTQDKE URQKNCIG KP OCK\G UKNCIG EQWNF DG FGNC[GF CPF VJG ITQYVJ QH URQKNCIG [GCUVU KP [QIJWTV YCU EQORNGVGN[ KPJKDKVGF D[ VJG CFFKVKQP QH VJG MKNNGT RTQVGKP ýLQYGU GV CN. ùøøøû. 6JG W. mrakii MKNNGT VQZKP YCU CNUQ RTQFWEGF KP S. cerevisiaeÿ WUKPI C OWNVK-EQR[ KPVGITCVKQP XGEVQT. 6JG TGEQODKPCPV [GCUV UVTCKP YCU CDNG VQ RTQFWEG MKNNGT VQZKP VJCV KPJKDKVGF C VGUV UVTCKP QH P. anomala. 6JKU CRRTQCEJ OKIJV NGCF VQ RTQFWEVKQP UVTCKPU VJCV ECP RTGXGPV URQKNCIG D[ YKNF [GCUVU KP HQQF RTQFWEVKQP ýöKOWTC GV CN. 199ôCû.
10.ý %QPElWUKQPU CPF QWVlQQM #PVKHWPICN [GCUVU JCXG DGGP WUGF CU DKQEQPVTQN CIGPVUÿ KPVGPVKQPCN QT PQVÿ HQT C XGT[ NQPI VKOG. *QYGXGTÿ TGUGCTEJ GHHQTVU D[ C DTQCF UEKGPVKHKE EQOOWPKV[ JCXG QEEWTTGF QPN[ FWTKPI VJG NCUV ùô [GCTU. &WTKPI VJKU VKOGÿ CP KORTGUUKXG FKXGTUKV[ QH DKQEQPVTQN QTICPKUOU CPF OGEJCPKUOU JCU DGGP FKUEQXGTGF. 0QV UWTRTKUKPIN[ÿ FWG VQ VJG CXCKNCDKNKV[ QH RQYGTHWN OQNGEWNCT OGVJQFUÿ KFGPVKHKECVKQP CPF RJ[NQIGPGVKE CPCN[UKU DGECOG VJG OCKP HKGNF HQT VJG WUG QH IGPGVKE OGVJQFU KP DKQEQPVTQN TGUGCTEJ. óGRQTVU EQPEGTPKPI VJG IGPGVKE DCEMITQWPF QH VJG DKQEQPVTQN CEVKXKV[ CTG UVKNN TCTG. 6JG IGPGVKEU QH UQOG DKQEQPVTQN QTICPKUOU YGTG KPXGUVKICVGF VQ C EGTVCKP FGITGG DGECWUG VJG[ JCXG QVJGT DKQVGEJPQNQIKECNN[ KORQTVCPV RTQRGTVKGU ýG.I. P. guilliermondiiû. þGCUVU YKVJ DKQEQPVTQN CEVKXKV[ CTG OCKPN[ PQP-EQPXGPVKQPCN CPF VJG MPQYNGFIG QH VJGKT IGPGVKE U[UVGOU KU D[ HCT NGUU FGXGNQRGF VJCP S. cerevisiaeÿ OCMKPI KV FKHHKEWNV VQ GUVCDNKUJ CPF WVKNKUG CRRTQRTKCVG OGVJQFU. 6JGTG CTGÿ JQYGXGTÿ C ITQYKPI PWODGT QH TGRQTVU CDQWV VJG KPVTQFWEVKQP QH IGPGVKE OGVJQFU KP DKQEQPVTQN U[UVGOU HQT OCPKRWNCVKQP QH VJGKT CEVKXKV[. 6JGTG KU PQ FQWDV VJCV IGPGVKE OGVJQFU KP VJG PGCT HWVWTG YKNN KORTQXG QWT MPQYNGFIG CDQWV DKQEQPVTQN CPF YKNN JGNR VQ QRVKOKUG VJG DKQEQPVTQN CRRTQCEJGU. òP VJG QVJGT JCPFÿ GZRNQTKPI IGPGVKE U[UVGOU QVJGT VJCP S. cerevisiae YKNN GPJCPEG QWT MPQYNGFIG CDQWV FKXGTUKV[ QH IGPGVKE U[UVGOU CPF YKNNÿ VJGTGHQTGÿ EQPVTKDWVG VQ VJG HWTVJGT FGXGNQROGPV QH [GCUV CRRNKGF IGPGVKEU.
#EMPQYlGFIGOGPVU 6JG HKPCPEKCN UWRRQTV HQT TGUGCTEJ QP VJG DKQEQPVTQN [GCUV Pichia anomala HTQO VJG (QWPFCVKQP HQT 5VTCVGIKE ©PXKTQPOGPVCN óGUGCTEJ ý/+56ó#û CPF VJG ©ñ-
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
319
HWPFGF RTQjGEV $+ò2ò56*#ó8©56 ýõLöô-%61999-ø1ø÷ôû KU ITCVGHWNN[ CEMPQYNGFIGF.
üGHGTGPEGU #DG öÿ öWUCMC +ÿ (WMWK 5 ý197ôû /QTRJQNQIKECN EJCPIG KP VJG GCTN[ UVCIGU QH VJG OCVKPI RTQEGUU QH Rhodosporidium toruloides. , $CEVGTKQN 1ùùí71ø-71ü #MCFC óÿ /KPQOK öÿ öCK ,ÿ þCOCUJKVC +ÿ /K[CMCYC 6ÿ (WMWK 5 ý19ü9û /WNVKRNG IGPGU EQFKPI HQT RTGEWTUQTU QH TJQFQVQTWEKPG Aÿ C HCTPGU[N RGRVKFG OCVKPI RJGTQOQPG QH VJG DCUKFKQO[EGVQWU [GCUV Rhodosporidium toruloides. /QN %GNN $KQN 9í3491-349ü #PFTGYU ,*ÿ 5RGCT ó0ÿ 0QTFJGKO ©8 ýùøøùû 2QRWNCVKQP DKQNQI[ QH Aureobasidium pullulans QP CRRNG NGCH UWTHCEGU. %CP , /KETQDKQN 4üíôøø-ô13 #TTCU ) ý199÷û /QFG QH CEVKQP QH CP KUQNCVG QH Candida famata KP DKQNQIKECN EQPVTQN QH Penicillium digitatum KP QTCPIG HTWKVU. 2QUVJCTXGUV $KQN 6GEJPQN üí191-19ü #TTCU )ÿ &G %KEEQ 8ÿ #TTW 5ÿ LKOC ) ý199üû $KQEQPVTQN D[ [GCUVU QH DNWG OQWNF QH EKVTWU HTWKVU CPF VJG OQFG QH CEVKQP QH CP KUQNCVG QH Pichia guilliermondii. , *QTVKE 5EK $KQVGEJPQN 73í413-41ü #UJKFC 5ÿ 5JKOC\CMK 6ÿ öKVCPQ öÿ *CTC 5 ý19ü3û 0GY MKNNGT VQZKP QH Hansenula mrakii. #ITKE $KQN %JGO 47íù9ô3-ù9ôô #XKU 6,ÿ $oNCPIGT óó ýùøøùû /GEJCPKUOU CPF OGCPU QH FGVGEVKQP QH DKQEQPVTQN CEVKXKV[ QH Pseudozyma [GCUVU CICKPUV RNCPV-RCVJQIGPKE HWPIK. (©/5 þGCUV óGU ùíô-ü #XKU 6,ÿ %CTQP 5,ÿ $QGMJQWV 6ÿ *COGNKP ó%ÿ $GNCPIGT óó ýùøø1Cû /QNGEWNCT CPF RJ[UKQNQIKECN CPCN[UKU QH VJG RQYFGT[ OKNFGY CPVCIQPKUV Pseudozyma flocculosa CPF TGNCVGF HWPIK. 2J[VQRCVJQNQI[ 91íù49-ùô4 #XKU 6,ÿ *COGNKP ó%ÿ $oNCPIGT óó ýùøø1Dû #RRTQCEJGU VQ OQNGEWNCT EJCTCEVGTK\CVKQP QH HWPICN DKQEQPVTQN CIGPVUí UQOG ECUG UVWFKGU. %CP , 2NCPV 2CVJQN ù3íü-1ù $CK (þÿ LKCPI *þÿ ,KC ,* ýùøøøû 6CZQPQOKE TGNCVKQPUJKRU COQPI VJG VCZC KP VJG Candida guilliermondii EQORNGZÿ CU TGXGCNGF D[ EQORCTCVKXG GNGEVTQRJQTGVKE MCT[QV[RKPI. +PV , 5[UV ©XQN /KETQDKQN ôøí417-4ùù $CK (þÿ <JCQ ,*ÿ 6CMCUJKOC /ÿ ,KC ,*ÿ $QGMJQWV 6ÿ 0CMCUG 6 ýùøøùû óGENCUUKHKECVKQP QH VJG Sporobolomyces roseus CPF Sporidiobolus pararoseus EQORNGZGUÿ YKVJ VJG FGUETKRVKQP QH Sporobolomyces phaffii UR. PQX. +PV , 5[UV ©XQN /KETQDKQN ôùíù3ø9-ù314 $CPPQ + ý19÷7û 5VWFKGU QH VJG UGZWCNKV[ QH Rhodotorula. , )GP #RRN /KETQDKQN 13í1÷7-19÷ $GEJGT &ÿ ${VVEJGT ( ý19ü3û 6JG EGNN V[RG QH Rhodosporidium toruloides CHVGT RTQVQRNCUV HWUKQP DGVYGGP UVTCKPU QH KFGPVKECN CPF QRRQUKVG OCVKPI V[RG. %WTT /KETQDKQN 9íù973øø $GIGTQY &ÿ $CWGT óÿ $QGMJQWV 6 ýùøøøû 2J[NQIGPGVKE RNCEGOGPVU QH WUVKNCIQO[EGVQWU CPCOQTRJU CU FGFWEGF HTQO PWENGCT L5ñ T&0# UGSWGPEGU. /[EQN óGU 1ø4íô3-÷ø $GP[CIQWD /ÿ $GN óJKNKF óÿ $oNCPIGT óó ý199÷Cû 2WTKHKECVKQP CPF EJCTCEVGTK\CVKQP QH PGY HCVV[ CEKFU YKVJ CPVKDKQVKE CEVKXKV[ RTQFWEGF D[ Sporothrix flocculosa. , %JGO ©EQN ùùí4øô-413 $GP[CIQWD /ÿ $oNCPIGT óó ý199ôû &GXGNQROGPV QH C OWVCPV UVTCKP QH Sporothrix flocculosa YKVJ TGUKUVCPEG VQ FQFGOQTRJ-CEGVCVG. 2J[VQRCVJQNQI[ üôí7÷÷-77ø $GP[CIQWD /ÿ úKNNGOQV %ÿ $oNCPIGT óó ý199÷Dû +PHNWGPEG QH C UWDKPJKDKVQT[ FQUG QH CPVKHWPICN HCVV[ CEKFU HTQO Sporothrix flocculosa QP EGNNWNCT NKRKF EQORQUKVKQP KP HWPIK. LKRKFU 31í1ø77-1øüù
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8QNMOCT 2CUUQVJ CPF ,QJCP 5EJP·TGT
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VTGCVOGPV HQT EQPVTQN QH RQUVJCTXGUV UVQTCIG TQVU QH HTWKVU CPF XGIGVCDNGU. $KQNQIKECN %QPVTQN 1øí9ü-1ø3 5W\WMK %ÿ 0KMMWPK 5 ý1994û 6JG RTKOCT[ CPF UWDWPKV UVTWEVWTG QH C PQXGN V[RG MKNNGT VQZKP RTQFWEGF D[ C JCNQVQNGTCPV [GCUVÿ Pichia farinosa. , $KQN %JGO ù÷9í3ø41-3ø4÷ 5W\WMK %ÿ þCOCFC öÿ òMCFC 0ÿ 0KMMWPK 5 ý19ü9û +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH JCNQVQNGTCPV MKNNGT [GCUVU HTQO HGTOGPVGF HQQFU. #ITKE $KQN %JGO ô3íùô93-ùô97 6GT LKPFG ,,ÿ 5VGGPUOC *þ ýùøøùû # OKETQCTTC[-CUUKUVGF UETGGP HQT RQVGPVKCN *CR1 CPF óQZ1 VCTIGV IGPGU KP Saccharomyces cerevisiae. þGCUV 1üíüùô-ü4ø 6JQTPGYGNN 5,ÿ 2GGT[ ó$ÿ 5MCVTWF 2L ý199ôû +PVGITCVKXG CPF TGRNKECVKXG IGPGVKE VTCPUHQTOCVKQP QH Aureobasidium pullulans. %WTT )GPGV ù9í÷÷-7ù 6KCP 52ÿ (CP õÿ :W þÿ ,KCPI #L ýùøøùCû ©HHGEVU QH ECNEKWO QP DKQEQPVTQN CEVKXKV[ QH [GCUV CPVCIQPKUVU CICKPUV VJG RQUVJCTXGUV HWPICN RCVJQIGP Rhizopus stolonifer. 2NCPV 2CVJQNQI[ ô1í3ôù-3ôü 6KCP 52ÿ (CP õÿ :W þÿ LKW *$ ýùøøùDû $KQEQPVTQN GHHKECE[ QH CPVCIQPKUV [GCUVU VQ ITC[ OQNF CPF DNWG OQNF QP CRRNGU CPF RGCTU KP EQPVTQNNGF CVOQURJGTGU. 2NCPV &KUGCUG ü÷íü4ü-üô3 6KRRGT &,ÿ $QUVKCP ö# ý19ü4û &QWDNG-UVTCPFGF TKDQPWENGKE CEKF MKNNGT U[UVGOU KP [GCUVU. /KETQDKQN óGX 4üí1ùô-1ô÷ 6WNN[ / ý19üôû ©PTKEJOGPV QH OWVCPVU QH Rhodosporidium toruloides D[ VJG WUG QH KPQUKVQN UVCTXCVKQP. , $CUKE /KETQDKQN ùôí÷ü3-÷ü÷ 6WNN[ /ÿ )KNDGTV *, ý19üôû 6TCPUHQTOCVKQP QH Rhodosporidium toruloides. )GPG 3÷íù3ôù4ø ñTSWJCTV ©,ÿ /GP\KGU ,)ÿ 2WPjC <ö ý1994û )TQYVJ CPF DKQNQIKECN EQPVTQN CEVKXKV[ QH Tilletiopsis URGEKGU CICKPUV RQYFGT[ OKNFGY ýSphaerotheca fuligineaû QP ITGGPJQWUG EWEWODGT. 2J[VQRCVJQNQI[ ü4í341-3ô1 ñTSWJCTV ©,ÿ 2WPjC <ö ýùøøùû *[FTQN[VKE GP\[OGU CPF CPVKHWPICN EQORQWPFU RTQFWEGF D[ Tilletiopsis URGEKGUÿ RJ[NNQURJGTG [GCUVU VJCV CTG CPVCIQPKUVU QH RQYFGT[ OKNFGY HWPIK. %CP , /KETQDKQN 4üíù19-ùù9 ñT\t %ÿ &G LGQ (ÿ LQ 2CUUQ %ÿ %TKUGQ ) ý1999û +PVTC-URGEKHKE FKXGTUKV[ QH Aureobasidium pullulans UVTCKPU KUQNCVGF HTQO TQEMU CPF QVJGT JCDKVCVU CUUGUUGF D[ RJ[UKQNQIKECN OGVJQFU CPF D[ TCPFQO CORNKHKGF RQN[OQTRJKE &0# ýó#2&û. , /KETQDKQN /GVJ 3÷í9ô-1øô 8CNGPVG 2ÿ óCOQU ,2ÿ LGQPEKPK ò ý1999û 5GSWGPEKPI CU C VQQN KP [GCUV OQNGEWNCT VCZQPQO[. %CP , /KETQDKQN 4ôí949-9ôü XCP FGT úCNV ,2ÿ 6C[NQT /$ÿ LKGDGPDGTI 08 ý1977û 2NQKF[ÿ CUEWU HQTOCVKQP CPF TGEQODKPCVKQP KP Torulaspora ýDebaryomycesû hansenii. #PVQPKG XCP LGGWYGPJQGM 43íùøôù1ü 8CPFGP ú[OGNGPDGTI #ÿ %WNNGP &ÿ 5RGCT ó0ÿ #PFTGYU ,* ý1999û óGIWNCVGF GZRTGUUKQP QH ITGGP HNWQTGUEGPV RTQVGKP WPFGT VJG EQPVTQN QH Aureobasidium pullulans Z[NCPCUG IGPG xylA. (©/5 /KETQDKQN LGVV 1ü1íùøô-ùø9 8CWIJCP-/CTVKPK #ÿ #PIGNKPK 2ÿ %CTFKPCNK ) ýùøøøû ñUG QH EQPXGPVKQPCN VCZQPQO[ÿ GNGEVTQRJQTGVKE MCT[QV[RKPI CPF &0#-&0# J[DTKFK\CVKQP HQT VJG ENCUUKHKECVKQP QH HGTOGPVCVKXG CRKEWNCVG [GCUVU. +PV , 5[UV ©XQN /KETQDKQN ôøí1÷÷ô-1÷7ù 8WUVKP //ÿ 5JGOjCMKPC 6/ÿ óGDGPVKUJ $#ÿ $[VUJMQXC /#ÿ öJCTKVQPQX 5+ÿ $GNjCjGX 58ÿ 6KOQMJKPC ©#ÿ 5KPGQMKj 52 ý19ü9û =öKNNGT RTQVGKPU RTQFWEGF D[ VJG [GCUV Hansenula anomala?. &QMN #MCF 0CWM 555ó 3øüí1ùô1-1ùôô ýKP óWUUKCPû úCNMGT )/ÿ /ENGQF #*ÿ *QFIUQP 8, ý199ôû +PVGTCEVKQPU DGVYGGP MKNNGT [GCUVU CPF RCVJQIGPKE HWPIK. (©/5 /KETQDKQN LGVV 1ù7íù13-ùùù
1ø 0QP-EQPXGPVKQPCN [GCUVU KP CPVKHWPICN CRRNKECVKQP
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úCPI 66ÿ %JQK þ,ÿ LGG $* ýùøø1û 6TCPUHQTOCVKQP U[UVGOU QH PQP-Saccharomyces [GCUVU. %TKV óGX $KQVGEJPQN ù1í177-ù1ü úGDGT *ÿ $CTVJ ) ý19üüû 0QPEQPXGPVKQPCN [GCUVUí VJGKT IGPGVKEU CPF DKQVGEJPQNQIKECN CRRNKECVKQPU. %TKV óGX $KQVGEJPQN 7íùü1-337 úGNUJ ,ÿ /E%NGNNCPF / ý199øû (KPIGTRTKPVKPI IGPQOGU WUKPI 2%ó YKVJ CTDKVTCT[ RTKOGTU. 0WENGKE #EKFU óGU 1üí7ù13-7ù1ü úGUQNQYUMK /ÿ (WMWJCTC * ý19ü1û LKPGCT OKVQEJQPFTKCN FGUQZ[TKDQPWENGKE CEKF HTQO VJG [GCUV Hansenula mrakii. /QN %GNN $KQN 1í3ü7-393 úKNNKCOU ,)öÿ öWDGNKM #óÿ LKXCM ö,ÿ óCHCNUMK ,#ÿ 6KPIG[ 58 ý199øû &0# RQN[OQTRJKUO CORNKHKGF D[ CTDKVTCT[ RTKOGTU CTG WUGHWN CU IGPGVKE OCTMGTU. 0WENGKE #EKFU óGU 1üí÷ô31-÷ô3ô úKNUQP %Lÿ ©NIJCQWVJ #ÿ %JCNWV\ ©ÿ &TQD[ 5ÿ 5VGXGPU %ÿ LW ,þÿ öJCP 8ÿ #TWN , ý1994û 2QVGPVKCN QH KPFWEGF TGUKUVCPEG VQ EQPVTQN RQUVJCTXGUV FKUGCUGU QH HTWKVU CPF XGIGVCDNGU. 2NCPV &KUGCUG 7üíü37-ü44 úKUPKGYUMK /ÿ $KNGU %ÿ &TQD[ 5ÿ /ENCWIJNKP óÿ úKNUQP %ÿ %JCNWV\ © ý1991û /QFG QH CEVKQP QH VJG RQUVJCTXGUV DKQEQPVTQN þGCUVÿ Pichia guilliermondii .1. %JCTCEVGTK\CVKQP QH CVVCEJOGPV VQ Botrytis cinerea. 2J[UKQN /QN 2NCPV 2 39íù4ô-ùôü þCOCOQVQ 6ÿ *KTCVCPK 6ÿ *KTCVC *ÿ +OCK /ÿ þCOCIWEJK * ý19ü÷û öKNNGT VQZKP HTQO *CPUGPWNC OTCMKK UGNGEVKXGN[ KPJKDKVU EGNN YCNN U[PVJGUKU KP C UGPUKVKXG [GCUV. (©$5 LGVV 197íôø-ô4 þCR 0#ÿ FG $CTTQU LQRGU /ÿ LCPITKFIG 2ÿ *GPUEJMG 2# ýùøøøû 6JG KPEKFGPEG QH MKNNGT CEVKXKV[ QH PQP-Saccharomyces [GCUVU VQYCTFU KPFKIGPQWU [GCUV URGEKGU QH ITCRG OWUVí RQVGPVKCN CRRNKECVKQP KP YKPG HGTOGPVCVKQP. , #RRN /KETQDKQN ü9í3ü1-3ü9 þCTYQQF %© ý193ùû Ampelomyces quisqualis QP ENQXGT OKNFGY. 2J[VQRCVJQNQI[ ùùí31 þGJWFC *ÿ &TQD[ 5ÿ úKUPKGYUMK /ÿ )QNFYC[ / ýùøø1û # VTCPUHQTOCVKQP U[UVGO HQT VJG DKQEQPVTQN [GCUVÿ Candida oleophilaÿ DCUGF QP J[ITQO[EKP $ TGUKUVCPEG. %WTT )GPGV 4øíùüù-ùü7 þQWPI 6úÿ þCIKW / ý197üû # EQORCTKUQP QH VJG MKNNGT EJCTCEVGT KP FKHHGTGPV [GCUVU CPF KVU ENCUUKHKECVKQP. #PVQPKG XCP LGGWYGPJQGM 44íô9-77
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG %JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
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
11.1 +PVTQFWEVKQP +P KPFWUVT[, VJGTG KU C EQPUVCPV UVTKXG VQ KORTQXG VJG RGTHQTOCPEG QH OCPWHCEVWTKPI RTQEGUUGU, CPF VJGTG KU IGPGTCNN[ HQEWU QP QRVKOKUCVKQP QH [KGNF, CPF/QT RTQFWEVKXKV[, CPF VQ CRRN[ CU EJGCR TCY OCVGTKCNU CU RQUUKDNG. 6JWU, KP C V[RKECN NKHG E[ENG VJG KPFWUVTKCN RTQEGUU KU EQPUVCPVN[ KORTQXGF TGUWNVKPI KP EJGCRGT RTQFWEVU þCV NGCUV QP C TGNCVKXG UECNG). 6QFC[, VJG EJGOKECN KPFWUVT[ KU VWTPKPI VQ DKQVGEJPQNQI[ HQT RTQFWEVKQP QH DQVJ HKPG EJGOKECNU CPF EQOOQFKV[ V[RG RTQFWEVU NKMG HWGNU, UQNXGPVU, CPF RQN[OGTU þ%JQVCPK GV CN. 2ýýý). 6JG MG[ FTKXGTU HQT VJKU CTG: 8“ %JGCRGT TCY OCVGTKCNU. 6JG TGNCVKXG EQUVU QH ECTDQJ[FTCVG TCY OCVGTKCNU CTG FGETGCUKPI FWG VQ KPVTQFWEVKQP QH OQTG GHHKEKGPV ETQRU CPF JCTXGUVKPI OGVJQFU. 8“ #DKNKV[ VQ RTQFWEG PQXGN EQORQWPFU. 6JTQWIJ DKQVGEJPQNQI[, EQORNGVGN[ PGY EQORQWPFU ECP DG OCPWHCEVWTGF CPF VJGUG OC[ HKPF CRRNKECVKQPU CU RJCTOCEGWVKECNU, HKPG EJGOKECNU CPF RQN[OGTU YKVJ KORTQXGF RTQRGTVKGU.
Topics in Current Genetics, Vol. 2 J.H. de Winde (Ed.): Functional Genetics of Industrial Yeasts Springer-Verlag Berlin Heidelberg 2003
332
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
8“ )TGGP EJGOKUVT[. $KQVGEJPQNQI[ TGRTGUGPVU C ITGGP VGEJPQNQI[, CPF GPXKTQPOGPVCN EQPUVTCKPVU KORQUGF QP VJG EJGOKECN KPFWUVT[ OCMG KV CVVTCEVKXG VQ HKPF CNVGTPCVKXG RTQFWEVKQP TQWVGU. 6JG YKFG CRRNKECVKQP QH DKQVGEJPQNQI[ KP VJG EJGOKECN KPFWUVT[ KU OCKPN[ FGVGTOKPGF D[ VJG CDKNKV[ VQ RGTHQTO FKTGEVGF IGPGVKE OQFKHKECVKQPU, CP CRRTQCEJ TGHGTTGF VQ CU OGVCDQNKE GPIKPGGTKPI. 6JKU VGTO JCU DGGP FGHKPGF CU OCMKPI TCVKQPCN KORTQXGOGPVU QH EGNNWNCT RTQEGUUGU VJTQWIJ KPVTQFWEVKQP QH FKTGEVGF IGPGVKE EJCPIGU D[ IGPGVKE GPIKPGGTKPI þ$CKNG[ 1991), CPF KV KPXQNXGU DQVJ FGUKIP QH PQXGN EGNN HCEVQTKGU CPF KORTQXGOGPV QH GZKUVKPI QPGU. 6TCFKVKQPCNN[, KORTQXGOGPVU QH C IKXGP EGNN HCEVQT[ YGTG UQWIJV D[ ENCUUKECN UVTCKP KORTQXGOGPV, YJKEJ KPXQNXG TCPFQO OWVCIGPGUKU QT ENCUUKECN DTGGFKPI QH VYQ UVTCKPU HQNNQYGF D[ UETGGPKPI HQT OWVCPVU QT J[DTKFU YKVJ VJG FGUKTGF RTQRGTVKGU þUGG %JCRVGT 2). &GURKVG VJG TCPFQO PCVWTG QH ENCUUKECN OGVJQFU, VJG[ JCXG DGGP SWKVG UWEEGUUHWN HQT KORTQXKPI VJG RGTHQTOCPEG QH OCP[ RTQFWEVKQP QTICPKUOU, CU KNNWUVTCVGF YKVJ VJG 1ý,ýýý HQNF KORTQXGOGPV KP RGPKEKNNKP RTQFWEVKXKV[ QDVCKPGF QXGT üý [GCTU QH ENCUUKECN UVTCKP KORTQXGOGPV þ0KGNUGP 2ýýý), CPF VJG[ UVKNN RNC[ C XGT[ KORQTVCPV TQNG KP KPFWUVT[. *QYGXGT, OGVCDQNKE GPIKPGGTKPI CNNQYU HQT KORTQXGOGPV QH VJG EGNN KP C OWEJ OQTG FKTGEVGF YC[, DGECWUG OQFKHKECVKQPU ECP DG VCTIGVGF VQ VJG RCVJYC[ QH KPVGTGUV. %QORCTGF VQ ENCUUKECN UVTCKP KORTQXGOGPV OGVCDQNKE GPIKPGGTKPI QHHGTU OCP[ QVJGT RQUUKDKNKVKGU CPF CFFKVKQPCN CFXCPVCIGU, G.I. KPVTQFWEVKQP QH JGVGTQNQIQWU IGPGU. 6JG URGEKHKE EJCPIGU KPVTQFWEGF CTG MPQYP, VJGTGHQTG KV KU RQUUKDNG VQ KORNGOGPV URGEKHKE KORTQXGOGPVU KP OCP[ FKHHGTGPV UVTCKPU QT QTICPKUOU CPF VJWU C UWEEGUUHWN UVTCVGI[ ECP DG CRRNKGF HQT KORTQXGOGPV QH FKHHGTGPV RTQEGUUGU. 5WEEGUUHWN OGVCDQNKE GPIKPGGTKPI UVCTVU YKVJ C ECTGHWN CPCN[UKU QH EGNNWNCT HWPEVKQP. $CUGF QP VJG CPCN[UKU, CP KORTQXGF UVTCKP KU FGUKIPGF CPF UWDUGSWGPVN[ EQPUVTWEVGF D[ IGPGVKE GPIKPGGTKPI þ0KGNUGP 2ýý1). +P QTFGT VQ GXCNWCVG VJG GPIKPGGTGF UVTCKP, KV KU PGEGUUCT[ VQ HQNNQY WR YKVJ CPCN[UKU, CPF VJKU OC[ RQKPV VQ PGY FGUKIP UVTCVGIKGU HQT KORTQXGOGPV QH VJG QTICPKUO. 6JGTGHQTG VJG RTQEGUU QH OGVCDQNKE GPIKPGGTKPI QHVGP KPXQNXGU C E[ENKE QRGTCVKQP þUGG (KIWTG 1) þ0KGNUGP 2ýý1) YJGTG UGXGTCN TQWPFU QH U[PVJGUKU CPF CPCN[UKU CTG TGSWKTGF VQ CEJKGXG VJG FGUKTGF KORTQXGOGPV QH VJG EGNN HCEVQT[. 6Q DG CDNG VQ TCVKQPCNN[ FGUKIP PGY UVTCVGIKGU HCUV CPF GHHKEKGPVN[, KV KU KORQTVCPV VJCV VJG RJGPQV[RKE EJCTCEVGTK\CVKQP þQT VJG CPCN[UKU RCTV) KU UWHHKEKGPVN[ FGVCKNGF VQ OCR URGEKHKE OGEJCPKUOU VJCV CTG KORQTVCPV HQT VJG RTQEGUU. 0GXGTVJGNGUU, KV KU VKOG EQPUWOKPI VQ RGTHQTO C FGVCKNGF RJGPQV[RKE EJCTCEVGTK\CVKQP, CPF KV KU VJGTGHQTG KORQTVCPV VJCV QPN[ TGNGXCPV CPCN[UKU QH VJG UVTCKP KU ECTTKGF QWV. &GRGPFKPI QP VJG OCPKRWNCVKQPU OCFG, C HGY ECTGHWNN[ UGNGEVGF CPCN[UKU VGEJPKSWGU, HTQO VJG VQQNDQZ QH CXCKNCDNG OGVJQFU, CTG PQTOCNN[ WUGF. ûDXKQWUN[, PQXGN JKIJ-VJTQWIJRWV VGEJPKSWGU FGXGNQRGF KP VJG HKGNF QH HWPEVKQPCN IGPQOKEU DGEQOG CVVTCEVKXG CU VJG[ CNNQY HQT CPCN[UKU QH OCP[ HCEVQTU KP C UKPING CPCN[UKU TWP. *QYGXGT, VJG[ CNUQ JCXG VJG IGPGTCN FTCYDCEM QH JKIJ-VJTQWIJRWV VGEJPKSWGU KP VJCV VJG[ CTG TGNCVKXGN[ KPUGPUKVKXG OGCUWTGOGPVU. 6JGTGHQTG, KV KU C RTGECTKQWU DCNCPEG VQ KFGPVKH[ OGVJQFU VJCV CNNQY HQT GNWEKFCVKQP QH VJG HCEVQTU CPF OGEJCPKUOU TGURQPUKDNG HQT VJG EQPVTQN QH OGVCDQNKE HNWZGU YJKEJ KU QHVGP TGSWKTGF HQT FGUKIP QH UWEEGUUHWN OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU HQT C IKXGP EGNN HCEVQT[.
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
333
(KI. 1. 6JG RTQEGUU QH OGVCDQNKE GPIKPGGTKPI. úGVCDQNKE GPIKPGGTKPI QHVGP KPXQNXGU C E[ENKE RTQEGUU UVCTVKPI KP QPG QH VJG VYQ DQZGU. 5[PVJGUKU QH C PGY OWVCPV D[ IGPGVKE GPIKPGGTKPI KU HQNNQYGF D[ CPCN[UKU QH RJGPQV[RG VQ SWCPVKH[ EJCPIGU CPF RQUUKDNG KORTQXGOGPVU. #PCN[UKU KU WUWCNN[ RGTHQTOGF YKVJ C HGY ECTGHWNN[ UGNGEVGF CPCN[UKU VQQNU HTQO VJG YKFG UGNGEVKQP QH VQQNU, CPF VJG CPCN[UKU VJGP OC[ RQKPV VQ PGY FGUKIP UVTCVGIKGU HQT KORTQXGOGPV QH VJG QTICPKUO. ûPG QHVGP JCU VQ VCMG UGXGTCN TQWPFU QH U[PVJGUKU CPF CPCN[UKU VQ CEJKGXG VJG FGUKTGF KORTQXGOGPV QH VJG EGNN HCEVQT[.
ùG YKNN FKUEWUU VJG KORCEV QH IGPQOKEU QP OGVCDQNKE GPIKPGGTKPI, CPF GURGEKCNN[, JQY VQQNU HTQO HWPEVKQPCN IGPQOKEU ECP DG WUGF VQ RGTHQTO XGT[ FGVCKNGF RJGPQV[RKE EJCTCEVGTKUCVKQP. 6QFC[, VJG NKOKVKPI EQORQPGPV QH VJG OGVCDQNKE GPIKPGGTKPI E[ENG þ(KIWTG 1) KU VJG TCVKQPCN CPCN[UKU QH EGNNWNCT RTQEGUUGU HQT VJG KFGPVKHKECVKQP QH VCTIGV IGPGU CPF GP\[OGU. *QYGXGT, VJG CFXCPEGU KP HWPEVKQPCN IGPQOKEU QHHGT OCP[ PGY VQQNU HQT INQDCN CPCN[UKU QH EGNNU CPF VJGKT UWEEGUUHWN CRRNKECVKQPU CTG VJGTGHQTG DQWPF VQ JCXG C JWIG KORCEV QP OGVCDQNKE GPIKPGGTKPI KP VJG PGCT HWVWTG þ%JQVCPK GV CN. 2ýýýø 5CPHQTF GV CN. 2ýý2).
11.2 ©ZCORNGU QH OGVCDQNKE GPIKPGGTKPI 6JGTG CTG OCP[ UWEEGUUHWN GZCORNGU QH OGVCDQNKE GPIKPGGTKPI DQVJ KP [GCUVU CPF QVJGT QTICPKUOU. úGVCDQNKE GPIKPGGTKPI JCU DGGP WUGF VQ KORTQXG RTQEGUUGU CRRNKGF HQT JGVGTQNQIQWU RTQVGKP RTQFWEVKQP CPF RTQFWEVKQP QH OGVCDQNKVGU D[ G.I. KP-
33ü
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
VTQFWEKPI PGY RCVJYC[U GPEQFGF D[ HQTGKIP IGPGU, GZVGPFKPI VJG UWDUVTCVG TCPIG, QT KORTQXKPI VJG [KGNF QT RTQFWEVKXKV[ þ0KGNUGP 2ýý1)þUGG CNUQ 6CDNG 1). ©URGEKCNN[ OGVCDQNKE GPIKPGGTKPI QH [GCUV JCU DGGP CP CTGC QH HQEWU FWG VQ VJG YKFG KPFWUVTKCN WUG QH Saccharomyces cerevisiae, CPF OCP[ GZCORNGU QH OGVCDQNKE GPIKPGGTKPI QH S. cerevisiae JCXG CRRGCTGF KP VJG NCUV FGECFG þûUVGTICCTF GV CN. 2ýýýD). +P VJKU EJCRVGT, YG YKNN IKXG C HGY GZCORNGU, YJKEJ YKNN EQXGT GZVGPUKQP QH UWDUVTCVG TCPIG, JGVGTQNQIQWU RTQVGKP RTQFWEVKQP, CPF TGFKTGEVKQP QH HNWZGU KP [GCUV. (QT OQTG GZCORNGU YG TGHGT VQ UQOG TGEGPV TGXKGYU þ0KGNUGP 2ýý1ø ûUVGTICCTF GV CN. 2ýýýD). 6CDNG 1. ©ZCORNGU QH OGVCDQNKE GPIKPGGTKPI KP [GCUV 6CTIGV ©ZVGPUKQP QH UWDUVTCVG TCPIG
öCVJYC[U NGCFKPI VQ PGY RTQFWEVU
*GVGTQNQIQWU RTQVGKP RTQFWEVKQP
+ORTQXGOGPV QH [KGNF CPF/QT RTQFWEVKXKV[
©NKOKPCVKQP QT TGFWEVKQP QH D[-RTQFWEV HQTOCVKQP +ORTQXGOGPV QH RTQEGUU RGTHQTOCPEG
©ZCORNG - S. cerevisiae YCU VTCPUHQTOGF YKVJ IGPGU GPEQFKPI Z[NQUG TGFWEVCUG CPF Z[NKVQN FGJ[FTQIGPCUG HTQO Pichia stipitis. 6JKU GPCDNGF S. cerevisiae VQ WVKNKUG Z[NQUG. - S. cerevisiae, VTCPUHQTOGF YKVJ VJG IGPG GPEQFKPI Z[NQUG TGFWEVCUG, RTQFWEGF Z[NKVQN HTQO Z[NQUG. - S. cerevisiae VTCPUHQTOGF YKVJ VJG IGPGU GPEQFKPI õ-OGVJ[NUCNKEWNKE CEKF U[PVJCUG HTQO Penicillium patulum CPF C JGVGTQNQIQWU RJQURJQRCPVGVJGKP[N VTCPUHGTCUG RTQFWEGU VJG RQN[MGVKFG õOGVJ[NUCNKE[NKE CEKF. - ;KGNF QH KPUWNKP UGETGVGF HTQO KPUWNKPRTQFWEKPI S. cerevisiae YCU KORTQXGF D[ WUKPI C U[PVJGVKE RTGRTQ-NGCFGT UGSWGPEG. - *KIJ NGXGN GZRTGUUKQP QH C UYGGV RTQVGKP, OQPGNNKP KP VJG HQQF [GCUV Candida utilis - +ORTQXKPI [KGNF QH GVJCPQN KP S. cerevisiae D[ FGNGVKPI VJG IGPG GPEQFKPI 0#&ö*-FGRGPFGPV INWVCOCVG FGJ[FTQIGPCUG CPF QXGT-GZRTGUUKPI VJG ô5ôûô#6 RCVJYC[ - 4GFWEKPI FKCEGV[N HQTOCVKQP KP DTGYGTU [GCUV D[ QXGT-GZRTGUUKPI VJG IGPG GPEQFKPI CEGVQNCEVCVG TGFWEVQKUQOGTCUG. - #NNGXKCVKQP QH INWEQUG EQPVTQN QP ICNCEVQUG OGVCDQNKUO KP CP KPFWUVTKCN UVTCKP QH S. cerevisiae D[ FGNGVKPI VJG IGPGU GPEQFKPI VJG PGICVKXG TGIWNCVQTU ôCN8ý CPF úKI1.
4GHGTGPEG þ÷{VVGT GV CN. 199ýø ÷{VVGT CPF %KTKCE[ 1993) þ*CNNDQTP GV CN. 1991) þ÷GCNG[ GV CN. 1998)
þ÷LGNFUGP GV CN. 1997) þ÷QPFQ GV CN. 1997) þ0KUUGP GV CN. 2ýýý)
þôLGTOCPUGP GV CN. 1988ø úKVJKGWZ CPF ùGKUU 199ó) þ4òPPQY GV CN. 1999)
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
33ó
11.2.1 ©ZVGPUKQP QH UWDUVTCVG TCPIG ñUWCNN[ EQUVU QH TCY OCVGTKCN EQPUVKVWVG C EQPUKFGTCDNG RCTV QH VJG VQVCN RTQFWEVKQP EQUVU KP KPFWUVTKCN HGTOGPVCVKQP RTQEGUUGU RTQFWEKPI NQY-XCNWG-CFFGF RTQFWEVU. 6JWU, KV KU CVVTCEVKXG VQ JCXG CU EJGCR C TCY OCVGTKCN CU RQUUKDNG KP UWEJ RTQEGUUGU. úQUV [GCUV UVTCKPU JCXG C PCTTQY UWDUVTCVG TCPIG, UQ VQ CEEQORNKUJ WVKNKUCVKQP QH C EJGCR TCY OCVGTKCN KV KU QHVGP PGEGUUCT[ VQ GKVJGT EJQQUG CPQVJGT QTICPKUO WUGF HQT RTQFWEVKQP QT GPCDNG KV VQ WUG VJG YCPVGF TCY OCVGTKCN. 6JKU CRRNKGU HQT GVJCPQN RTQFWEVKQP KP S. cerevisiae KP YJKEJ VJGTG KU C UWDUVCPVKCN KPVGTGUV VQ CRRN[ EJGCRGT HGGFUVQEMU UWEJ CU NKIPQEGNNWNQUKE OCVGTKCNU FGTKXGF HTQO CITKEWNVWTCN TGUKFWGU. +P QTFGT VQ FQ VJKU, KV KU TGSWKTGF VJCV S. cerevisiae GHHGEVKXGN[ ECP EQPXGTV RGPVQUGU NKMG Z[NQUG CPF CTCDKPQUG KPVQ GVJCPQN þ#TKUVKFQW CPF öGPVVKNk 2ýýýø *CJP*kIGTFCN GV CN. 2ýý1ø
33õ
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
(KI. 2. :[NQUG WVKNKUCVKQP KP GPIKPGGTGF S. cervisisiae. ùKNF V[RG S. cerevisiae KU PQV CDNG VQ WVKNKUG Z[NQUG, DWV GZRTGUUKQP QH Z[NQUG TGFWEVCUG þð4) CPF Z[NKVQN FGJ[FTQIGPCUG þð&*) HTQO P. stipitis GPCDNGU S. cerevisiae VQ ITQY QP Z[NQUG CU VJG UQNG ECTDQP UQWTEG þ÷{VVGT GV CN. 199ýø ÷{VVGT CPF %KTKCE[ 1993). ûXGT-GZRTGUUKQP QH VJG JQOQNQIQWU Z[NWNQUG MKPCUG þð÷) TGUWNVU KP CP KPETGCUGF GVJCPQN [KGNF CPF RTQFWEVKXKV[ þ*Q GV CN. 1998ø ÿQJCPUUQP GV CN. 2ýý1).
RTQVGKPU CTG VJGTGHQTG RQUUKDNG D[ OCPKRWNCVKPI VJG HQNFKPI GPXKTQPOGPV QH VJG ©4. 6JG *UR7ý EJCRGTQPG $Kö þDKPFKPI RTQVGKP) CRRGCTU VQ RNC[ C MG[ TQNG KP VJG OGEJCPKUO QH OQXGOGPV QH C RQN[RGRVKFG CETQUU VJG ©4 þï[OCP CPF 5EJGMOCP 1997ø öCP\PGT GV CN. 199óø 5EJCV\ CPF &QDDGTUVGKP 199õ), ©4 TGVGPVKQP þ&QTPGT CPF ÷CWHOCP 199ü), CU YGNN CU DKPFKPI VQ WPHQNFGF CPF WPCUUGODNGF RTQVGKPU KP VJG ©4 þ*CCU CPF ùCDN 1983). +V YCU RTGFKEVGF VJCV QXGT-GZRTGUUKQP QH $Kö YQWNF JCXG C RQUKVKXG TQNG HQT UGETGVKQP QH JGVGTQNQIQWU RTQVGKPU þ4QDKPUQP CPF ùKVVTWR 1993), DWV KV QPN[ FQGU UQ HQT UQOG JGVGTQNQIQWU RTQVGKPU. 5GETGVKQP QH UKPINGEJCKP CPVKDQF[ HTCIOGPVU KP S. cerevisiae DGPGHKVU HTQO VJG EJCRGTQPG HWPEVKQP QH $Kö QXGT-GZRTGUUKQP þ5JWUVC GV CN. 1998) D[ C OGEJCPKUO PQV UKIPKHKECPV HQT VJTGG QVJGT OQFGN RTQVGKPU UVWFKGF þ4QDKPUQP GV CN. 199õ). # DGVVGT WPFGTUVCPFKPI QH VJG OGEJCPKUOU VJCV KORTQXG JGVGTQNQIQWU RTQVGKP RTQFWEVKQP KU VJGTGHQTG PGGFGF, HQT GZCORNG VJTQWIJ UVWFKGU QH GZRTGUUKQP QH IGPGU TGNCVGF VQ U[PVJGUKU, HQNFKPI CPF UGETGVKQP QH RTQVGKPU, CU YGNN CU VJG GZCEV HWPEVKQP QH VJG IGPG RTQFWEVU. 11.2.3 +ORTQXGOGPV QH HNWZGU +P QTFGT VQ OCZKOKUG [KGNF CPF/QT RTQFWEVKXKV[, KV KU QHVGP FGUKTCDNG VQ KORTQXG HNWZGU, G.I. KPETGCUG QT FGETGCUG C IKXGP HNWZ. 6JKU ECP DG FQPG D[ KPETGCUKPI UWDUVTCVG WRVCMG QT TGFKTGEVKPI HNWZGU, CPF VJGTG CTG UGXGTCN GZCORNGU QH VJKU KP [GCUV, G.I. KPETGCUKPI HNWZ VQ GVJCPQN YJKNG FGETGCUKPI HNWZ VQ IN[EGTQN þ0KUUGP GV CN. 2ýýý) CPF TGFWEKPI FKCEGV[N HQTOCVKQP KP DTGYGT•U [GCUV þôLGTOCPUGP GV CN. 1988ø úKVJKGWZ CPF ùGKUU 199ó). +P S. cerevisiae, VJG ICNCEVQUG HNWZ KU VJTGG HQNF NQYGT VJCP VJCV QH INWEQUG þûUVGTICCTF GV CN. 2ýýýE), CPF VJGTGHQTG, KV KU CVVTCEVKXG VQ KP-
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
337
ETGCUG VJG ICNCEVQUG HNWZ HTQO CP KPFWUVTKCN RQKPV QH XKGY UKPEG UGXGTCN KPFWUVTKCN OGFKC EQPVCKPU ICNCEVQUG þûUVGTICCTF GV CN. 2ýýýEø ùKUGNQIGN GV CN. 199õ). 6JG ICNCEVQUG WVKNKUCVKQP RCVJYC[ EQPUKUVU QH HKXG GP\[OGU EQPXGTVKPI GZVTCEGNNWNCT ICNCEVQUG KPVQ INWEQUG-õ-RJQRJCVG, YJKEJ VJGP GPVGTU VJG IN[EQN[UKU þUGG (KIWTG 3). ûXGT-GZRTGUUKQP QH VJG IGPGU GPEQFKPI VJG HKTUV HQWT GP\[OGU KP VJG ICNCEVQUG WVKNKUCVKQP RCVJYC[, GKVJGT CNQPG QT KP EQODKPCVKQPU, TGUWNVU KP C FGETGCUGF ICNCEVQUG HNWZ þûUVGTICCTF 2ýýý). *QYGXGT, YJGP VJG IGPGU GPEQFKPI VJG PGICVKXG
(KI. 3. 6JG ICNCEVQUG WVKNKUCVKQP RCVJYC[ KP S. cerevisiae. ©ZVTCEGNNWNCT ICNCEVQUG KU VTCPURQTVGF KPVQ VJG EGNN D[ ICNCEVQUG RGTOGCUG þôCN2) CPF VJGP EQPXGTVGF VQ INWEQUG-õRJQURJCVG D[ TGCEVKQPU ECVCN[UGF D[ ICNCEVQMKPCUG þôCN1), ICNCEVQUG-1-RJQURJCVG WTKF[N[NVTCPUHGTCUG þôCN7), ñ&ö-INWEQUG ü-GRKOGTCUG þôCN1ý) CPF RJQURJQINWEQOWVCUG þôCNó). ôCNü CEVU CU C VTCPUETKRVKQPCN CEVKXCVQT QH VJG GAL IGPGU D[ DKPFKPI VQ URGEKHKE WRUVTGCO CEVKXCVKPI UKVGU þñ#5ô). ôCNCEVQUG CEVU CU C UKIPCN OQNGEWNG, YJKEJ þD[ CP #6ö-FGRGPFGPV OGEJCPKUO) KU DQWPF VQ ôCN3 VJCV CEVU CU C VTCPUFWEGT HQT ICNCEVQUG KPFWEVKQP D[ KPVGTCEVKPI YKVJ ôCN8ý CPF VJGTGD[ TGNKGXKPI ôCN8ý•U TGRTGUUKQP QH ôCNü. úKI1-OGFKCVGF EQPVTQN RTGXGPVU VTCPUETKRVKQP QH VJG GAL1 IGPG CPF VJG GAL4 IGPG KP VJG RTGUGPEG QH INWEQUG D[ RCTVKEKRCVKPI KP C RTQVGKP EQORNGZ YKVJ 5UPõ CPF 6WR1 VJCV DKPFU VQ URGEKHKE WRUVTGCO TGRTGUUKQP UKVGU þñ45). (WTVJGTOQTG, ôCNõ CNUQ FQYP-TGIWNCVGU VJG GAL U[UVGO D[ CHHGEVKPI RTQFWEVKQP, UVCDKNKUCVKQP, QT FGITCFCVKQP QH GAL1, GAL2 CPF GAL7 O40#, DWV VJG GZCEV OGEJCPKUO KU PQV YGNN MPQYP. ûUVGTICCTF GV CN. þ2ýýýC) KPETGCUGF VJG HNWZ VJTQWIJ VJG ICNCEVQUG WVKNKUCVKQP RCVJYC[ D[ QXGT-GZRTGUUKPI GAL4 QT FGNGVKPI GAL6, GAL80, CPF MIG1. (KIWTG OQFKHKGF HTQO ûUVGTICCTF GV CN. þ2ýýýC).
338
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
TGIWNCVQT[ RTQVGKPU ôCN8ý, ôCNõ, CPF úKI1 þUGG (KIWTG 3) YGTG FGNGVGF QT VJG VTCPUETKRVKQPCN CEVKXCVQT QH VJG ICNCEVQUG IGPGU, ôCNü, YCU QXGT-GZRTGUUGF, VJGTG YCU C RQUKVKXG GHHGEV CPF VJG ICNCEVQUG HNWZ KPETGCUGF YKVJ WR VQ üýî KP VJG VTKRNG FGNGVKQP OWVCPV þûUVGTICCTF GV CN. 2ýýýC). 6JKU KNNWUVTCVGU VJCV TGIWNCVQT[ RCVJYC[U CTG CVVTCEVCDNG VCTIGVU HQT OGVCDQNKE GPIKPGGTKPI CPF VJCV OWNVKRNG IGPGVKE OQFKHKECVKQPU ECP DG PGEGUUCT[ VQ QDVCKP DGVVGT KORTQXGOGPVU. $WV RQVGPVKCNN[, VJG ICNCEVQUG HNWZ ECP DG KPETGCUGF OWEJ HWTVJGT KH QPG CUUWOGU KV ECP DG DTQWIJV VQ VJG UCOG NGXGN CU VJG INWEQUG HNWZ. 6Q HWTVJGT TCVKQPCNN[ GPIKPGGT VJG ICNCEVQUG OGVCDQNKUO, C DGVVGT WPFGTUVCPFKPI QH VJG U[UVGO CPF C DGVVGT CDKNKV[ VQ RTQRGT UWTXG[ CNN VJG GHHGEVU QH VJG UWEEGUUHWN IGPGVKE OCPKRWNCVKQPU CTG PGEGUUCT[. #NN GHHGEVU QH IGPG OCPKRWNCVKQPU CTG HCT HTQO RTGFKEVCDNG, GXGP HQT C TGIWNCVQT[ U[UVGO UQ TGNCVKXGN[ YGNN WPFGTUVQQF CU VJCV QH VJG ICNCEVQUG WVKNKUCVKQP U[UVGO.
11.3 %JCNNGPIGU KP OGVCDQNKE GPIKPGGTKPI # HWPFCOGPVCN QDLGEVKXG QH OGVCDQNKE GPIKPGGTKPI KU VQ GNWEKFCVG VJG HCEVQTU CPF OGEJCPKUOU TGURQPUKDNG HQT VJG EQPVTQN QH OGVCDQNKE HNWZ, CPF VQ ICKP C DGVVGT WPFGTUVCPFKPI QH VJGUG HQTOU VJG DCUKU QH DGVVGT UVTCVGIKGU HQT OGVCDQNKE GPIKPGGTKPI. 6JG OGVCDQNKE HNWZGU CPF VJGKT EQPVTQN ECP DG CUUGUUGF D[ XCTKQWU VJGQTGVKECN OGVJQFU DCUGF QP RJ[UKQNQIKECN FCVC þ5VGRJCPQRQWNQU GV CN. 1998), DWV VJGUG VGEJPKSWGU CTG WUWCNN[ DCUGF QP UKORNKHKECVKQPU, URGEKHKE CUUWORVKQPU, CPF KPCFGSWCVG MPQYNGFIG, CPF VJG[ FQ PQV VCMG KPVQ CEEQWPV VJG OCP[ NGXGNU QH TGIWNCVKQP KP VJG EGNN. úGVCDQNKE EQPVTQN CPCN[UKU ECP DG C IQQF UVCTVKPI RQKPV, DWV QHVGP OCPKRWNCVKQP QH C IGPG, RTGFKEVGF VQ JCXG NCTIG EQPVTQN QH HNWZ VJTQWIJ C IKXGP RCVJYC[, JCU PQ RQUKVKXG GHHGEV CU KNNWUVTCVGF YKVJ VJG GHHQTVU VQ KPETGCUG VJG HNWZ VJTQWIJ IN[EQN[UKU þ5EJCCHH GV CN. 1989ø 5OKVU GV CN. 2ýýý). #P[ RGTVWTDCVKQP QH VJG IGPQOG GXGP C UKPING IGPGVKE EJCPIG, WUWCNN[ TGUWNVU KP C OWNVKIGPG/OWNVKVTCPUETKRV/OWNVKRTQVGKP TGURQPUG þ$CKNG[ 1999), YJKEJ OCMGU KV FKHHKEWNV VQ RTGFKEV CPF WPFGTUVCPF VJG GHHGEVU QH URGEKHKE IGPGVKE OQFKHKECVKQPU. ôNQDCN TGURQPUGU CTG CNYC[U KPXQNXGF CPF CNYC[U RQVGPVKCNN[ KORQTVCPV þ$CKNG[ 1999) CPF VJGTGHQTG, KV UGGOU KORQUUKDNG VQ EJCPIG VJG TGNCVKXG NGXGN QH QPN[ QPG VTCPUETKRV YKVJ CNN QVJGTU HKZGF. 6JKU KU C EQPUGSWGPEG QH VJG EQORNGZ TGIWNCVQT[ U[UVGOU RTGXCKNKPI KP VJG EGNN, CPF VJG TGFWPFCPE[ KP EQPVTQN QH OCP[ RCVJYC[U. #U C EQPUGSWGPEG QH GXQNWVKQP, VJGUG TGIWNCVQT[ U[UVGOU JCXG DGGP FGUKIPGF UWEJ VJCV VJG QXGTCNN EGNN HWPEVKQP KU TGNCVKXGN[ KPUGPUKVKXG VQ OWVCVKQPU CPF VJGTGHQTG OCP[ IGPGU, UKIPCNU, CPF TGIWNCVQT[ KPVGTCEVKQPU HCKN VQ JCXG CP[ UKIPKHKECPV GHHGEV QP RJGPQV[RG WPNGUU QVJGT IGPGU CTG UKOWNVCPGQWUN[ CNVGTGF. 6JG HNWZGU KP C EGNN CTG EQPVTQNNGF CV OCP[ NGXGNU þUGG (KIWTG ü), G.I. VTCPUETKRVKQPCN EQPVTQN, EQPVTQN QH O40# FGITCFCVKQP, VTCPUNCVKQPCN EQPVTQN, RTQVGKP CEVKXCVKQP/KPCEVKXCVKQP, CPF CNNQUVGTKE EQPVTQN QH GP\[OGU þ÷NGKP GV CN. 1998). 6Q KNNWUVTCVG VJG EQORNGZ TGIWNCVKQP KP VJG EGNN, YG OC[ URGEKH[ GCEJ QH VJG ÂQOGU• KP VJG EGNN CU HWPEVKQP QH VJG QVJGT ÂQOGU•: 6TCPUETKRVQOG = HVTCPUETKRVQOGþIGPQOG, RTQVGQOG, KPVGTCEVQOG) öTQVGQOG = HRTQVGQOGþVTCPUETKRVQOG, RTQVGQOG, KPVGTCEVQOG)
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
339
+PVGTCEVQOG = HKPVGTCEVQOGþIGPQOG, RTQVGQOG, OGVCDQNQOG) úGVCDQNQOG = HOGVCDQNQOGþHNWZQOG) (NWZQOG = HHNWZQOGþRTQVGQOG, OGVCDQNQOG) %NGCTN[, VJG RTQVGQOG RNC[U C MG[ TQNG KP VJG QXGTCNN TGIWNCVKQP, DWV VJG UGV QH HWPEVKQPU URGEKHKGF CDQXG KPFKECVGU VJG ENQUG KPVGTCEVKQP VJCV GZKUVU DGVYGGP VJG FKHHGTGPV ÂQOGU•. 6JWU, GXGP C UKPING IGPGVKE EJCPIG OKIJV G.I. XKC EJCPIGF OGVCDQNKVG EQPEGPVTCVKQPU KPHNWGPEG TGIWNCVQT[ RTQVGKPU NGCFKPI VQ WPHQTGUGGP UGEQPFCT[ GHHGEVU QP DQVJ VJG VTCPUETKRVQOG CPF RTQVGQOG. $GECWUG KV KU XGT[ FKHHKEWNV VQ RTGFKEV VJG EQORNGVG GHHGEV QP C IGPGVKE EJCPIG, KV KU QHVGP PGEGUUCT[ VQ IQ VJTQWIJ VJG E[ENG QH OGVCDQNKE GPIKPGGTKPI UGXGTCN VKOGU. +V JCU CNTGCF[ DGGP UGGP KP GZCORNGU JQY OWNVKRNG IGPGVKE OQFKHKECVKQPU ECP JGNR VQ DGVVGT GPIKPGGT UVTCKPU VQ CVVCKP FGUKTGF RJGPQV[RGU þûUVGTICCTF GV CN. 2ýýýC). 5WKVCDNG EQ-QTFKPCVGF GZRTGUUKQP þCPF/QT KPJKDKVKQP) QH UGXGTCN IGPGU YKNN VJGTGHQTG DG PGEGUUCT[ VQ CEJKGXG OQTG CFXCPEGF QDLGEVKXGU KP OGVCDQNKE GPIKPGGTKPI þöCRQWVUCMKU 1998), DWV KV KORQUGU C ITGCVGT FGOCPF QP QWT MPQYNGFIG QH VJG EGNN.
(KI. ÿ. *KGTCTEJKECN EQPVTQN QH HNWZ KP C EGNN. 6JG IGPGU KP C IGPQOG CTG VTCPUETKDGF KPVQ O40#U þVJG VTCPUETKRVQOG) VJCV CTG HWTVJGT VTCPUNCVGF KPVQ RTQVGKPU þVJG RTQVGQOG). 6JG RTQVGKPU FGVGTOKPG EGNNWNCT HWPEVKQP GKVJGT VJTQWIJ KPVGTCEVKQPU YKVJ RTQVGKP, &0# QT OGVCDQNKVGU þVJG KPVGTCEVQOG) QT VJTQWIJ VJGKT CEVKQP CU GP\[OGU, YJKEJ ECVCN[UG VJG KPVGTEQPXGTUKQP QH OGVCDQNKVGU þVJG OGVCDQNQOG). 6JG KPVGT-EQPXGTUKQP QH OGVCDQNKVGU EQPUVKVWVGU VJG OGVCDQNKE PGVYQTM CPF ECP DG SWCPVKHKGF CU HNWZGU þVJG HNWZQOG). 6JG OCP[ NGXGNU QH EQPVTQN OCMG OCRRKPI QH EGNNWNCT HWPEVKQPU FKHHKEWNV. 5[ODQNU: 5, UWDUVTCVGø X, KPVTCEGNNWNCT OGVCDQNKVGø ö, RTQFWEVø X, HNWZ.
3üý
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
4GIWNCVQT[ PGVYQTMU CTG RTQOKUKPI VCTIGVU HQT OGVCDQNKE GPIKPGGTKPI UKPEG OCPKRWNCVKQP QH C TGIWNCVQT[ U[UVGO OC[ CNNQY TGNGCUG QH QXGTCNN EQPVTQN QH C YJQNG RCVJYC[. *QYGXGT, VJG EQORNGVG TGIWNCVQT[ UVTWEVWTG KU QPN[ MPQYP HQT TGNCVKXGN[ HGY TGIWNCVQT[ U[UVGOU, CPF VJGTGHQTG, OGVCDQNKE GPIKPGGTKPI QHVGP KPXQNXGU UVWFKGU QP TGIWNCVQT[ PGVYQTMU KP QTFGT VQ KFGPVKH[ PGY RQUUKDNG VCTIGVU. ùJGP VCTIGVKPI TGIWNCVQT[ RCVJYC[U, WPGZRGEVGF GHHGEVU CTG QHVGP QDUGTXGF, UKPEG TGIWNCVQT[ RCVJYC[U OC[ TGIWNCVG QVJGT IGPGU VJCP VJG QPGU QH KPVGTGUV. #TQWPF QPG VJKTF QH S. cerevisiae QRGP TGCFKPI HTCOGU þû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cerevisiae. +P QTFGT VQ FGXGNQR DGVVGT OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU, KV KU RKXQVCN VJCV VJG GPVKTG EGNN KU EQPUKFGTGF CPF PQV LWUV UKPING RCVJYC[U QT GP\[OCVKE TGCEVKQPU. +P VJG RCUV, VJGTG JCU DGGP C VGPFGPE[ VQ HQEWU QP UKPING RCVJYC[U CPF UKPING GP\[OCVKE TGCEVKQPU, DWV EWTTGPVN[ VJGTG KU C RCTCFKIO UJKHV CYC[ HTQO KPFKXKFWCN GP\[OCVKE TGCEVKQPU VQYCTF U[UVGOU QH KPVGTCEVKPI DKQEJGOKECN TGCEVKQPU þ$TGPV 2ýýýø 0KGNUGP CPF ûNUUQP 2ýý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ÂQOGU•, YKNN JCXG C UKIPKHKECPV KORCEV QP OGVCDQNKE GPIKPGGTKPI. úCP[ QH VJGUG JKIJ-VJTQWIJRWV CPCN[UKU VGEJPKSWGU HKPF ITGCV WUG KP OGVCDQNKE GPIKPGGTKPI D[ GPCDNKPI C U[UVGOU CRRTQCEJ HQT UVWF[KPI VJG EGNN CU OGPVKQPGF CDQXG, DWV VJG[ OC[ CNUQ QRGP PGY HKGNFU QH OGVCDQNKE GPIKPGGTKPI UWEJ CU KPXGTUG OGVCDQNKE GPIKPGGTKPI þ$CKNG[ GV CN. 199õ).
11.4 (WPEVKQPCN IGPQOKEU 6JG IQCN QH HWPEVKQPCN IGPQOKEU KU VQ CUUKIP HWPEVKQP VQ GCEJ IGPG YKVJKP C IGPQOG. #UUKIPOGPV QH HWPEVKQP VQ CNN IGPGU PGEGUUKVCVGU C U[UVGOU CRRTQCEJ DQVJ OGPVCNN[ CPF CPCN[VKECNN[. +P VJG RCUV FGECFG, VJGTG JCU DGGP C VTGOGPFQWU FGXGNQROGPV QH PQXGN VGEJPKSWGU HQT CPCN[UKU QH VJG IGPQOG, VTCPUETKRVQOG, RTQVGQOG, KPVGTCEVQOG, CPF OGVCDQNQOG QP C YJQNG EGNN NGXGN. ;GCUVU, CPF RCTVKEWNCTN[ S. cerevisiae, JCXG C WPKSWG RQUKVKQP KP HWPEVKQPCN IGPQOKEU FWG VQ VJG TQNG QH S. cerevisiae CU C OQFGN QTICPKUO CPF DGKPI QPG QH VJG OQUV YGNN-UVWFKGF QTICPKUOU QH
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ü1
CNN. +V IQV C JGCF UVCTV YKVJ VJG RWDNKECVKQP QH KVU IGPQOG UGSWGPEG þôQHHGCW GV CN. 199õ) CU VJG HKTUV GWMCT[QVKE QTICPKUO CPF S. cerevisiae JCU DGGP C RKQPGGT KP CNN PGY VGEJPKSWGU KP HWPEVKQPCN IGPQOKEU GXGT UKPEG, K.G. YKVJ GZVGPUKXG UVWFKGU QH VJG VTCPUETKRVQOG þ&G4KUK GV CN. 1997ø íGNEWNGUEW GV CN. 1997ø ùQFKEMC GV CN. 1997), VJG RTQVGQOG þ(WVEJGT GV CN. 1999ø ô[IK GV CN. 1999ø 5JGXEJGPMQ GV CN. 199õ), VJG KPVGTCEVQOG þ+VQ GV CN. 2ýýýø +[GT GV CN. 2ýý1ø 4GP GV CN. 2ýýýø ñGV\ GV CN. 2ýýý), CPF VJG OGVCDQNQOG þ4CCOUFQPM GV CN. 2ýý1) þXK\. (KIWTG ü). (WPEVKQPCN IGPQOKEU QP [GCUVU YKNN VJGTGHQTG KPGXKVCDNG DTKPI QPG VQ HQEWU QP S. cerevisiae CNOQUV GPVKTGN[, DWV CU OQTG [GCUV IGPQOGU CTG UGSWGPEGF, VJKU YKNN TCRKFN[ EJCPIG. 11.4.1 %QORCTCVKXG UGSWGPEG CPCN[UKU 6JGTG YCU C NQPI ICR CHVGT VJG RWDNKECVKQP QH VJG S. cerevisiae IGPQOG KP 199õ DGHQTG RWDNKECVKQP QH QVJGT [GCUV IGPQOGU. &WG VQ VJKU, VJG HKGNF QH NCTIG-UECNG EQORCTCVKXG UGSWGPEKPI QH [GCUVU KU UVKNN PGY. $WV YKVJ VJG RWDNKECVKQP QH VJG IGPQOG UGSWGPEG QH VJG HKUUKQP [GCUV Schizosaccharomyces pombe þùQQF GV CN. 2ýý2) CPF VJG QPIQKPI UGSWGPEKPI QH OCP[ QVJGT [GCUVU þJVVR://YKV.KPVGITCVGFIGPQOKEU.EQO /ôûï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þ>OKV\CMKU GV CN. 2ýý2ø ùCVGTUVQP GV CN. 2ýý2). # HKPG GZCORNG KP [GCUV KU VJG ôoPQNGXWTGU RTQLGEV YJKEJ KU C NCTIG-UECNG EQORCTCVKXG RTQLGEV DGVYGGP S. cerevisiae CPF 13 QVJGT [GCUV URGEKGU TGRTGUGPVCVKXG QH VJG XCTKQWU DTCPEJGU QH VJG *GOKCUEQO[EGVQWU ENCUU þUGG þ5QWEKGV GV CN. 2ýýý) CPF TGHGTGPEGU JGTGKP). 6JKU RTQLGEV JCU RCTVN[ UGSWGPEGF VJG 13 [GCUV URGEKGU CPF KFGPVKHKGF CRRTQZKOCVGN[ 2ý,ýýý RTQVGKP-EQFKPI UGSWGPEGU KP VJG URGEKGU. +FGPVKHKECVKQP QH HWPEVKQPCN UGSWGPEG GNGOGPVU PQV EQFKPI HQT RTQVGKP, G.I. UGSWGPEGU TGIWNCVKPI IGPG GZRTGUUKQP, KU OWEJ OQTG EQORNKECVGF FWG VQ VJG TGNCVKXG TCRKF TCVG QH EJCPIG QH PQPRTQVGKP-EQFKPI UGSWGPEGU KP GXQNWVKQP CPF VJGKT TGNCVKXG UKORNG UVTWEVWTGU þ%NKHVGP GV CN. 2ýý1). %QORCTKUQP QH TGNCVKXG ENQUGN[ TGNCVGF QTICPKUOU KU OQTG NKMGN[ VQ KFGPVKH[ RQVGPVKCN PQPRTQVGKP-EQFKPI GNGOGPVU. 6JKU YCU VGUVGF D[ %NKHVGP GV CN. þ2ýý1), YJQ FKF EQORCTCVKXG UGSWGPEG CPCN[UKU QH GKIJV Saccharomyces URGEKGU CPF UJQYGF VJCV VJKU OGVJQF ECP DG UWEEGUUHWN. +P VJG PGCT HWVWTG, YJGP OQTG ENQUGN[ TGNCVGF URGEKGU CTG UGSWGPEGF, YG OKIJV UGG C FKUEQXGT[ QH RQVGPVKCNN[ PQXGN TGIWNCVQT[ &0# GNGOGPVU D[ EQORCTCVKXG UGSWGPEG CPCN[UKU.
3ü2
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
11.4.2 6TCPUETKRVQOG CPCN[UKU 6JG CDKNKV[ VQ RGTHQTO IGPQOG-YKFG VTCPUETKRVKQP CPCN[UKU YCU C OCLQT DTGCMVJTQWIJ KP HWPEVKQPCN IGPQOKEU. 6JG UGSWGPEKPI QH VJG S. cerevisiae IGPQOG VQIGVJGT YKVJ VGEJPKECN CEJKGXGOGPVU GPCDNGF VJG HKTUV &0# CTTC[U HQT IGPQOG-YKFG VTCPUETKRVKQP CPCN[UKU QH S. cerevisiae þCPF QH CP[ UGSWGPEGF QTICPKUO) KP VJG NCVG199ý•U þ&G4KUK GV CN. 1997ø ùQFKEMC GV CN. 1997). úCP[ QRVKQPU QH &0# CTTC[U GZKUV þ*GNNGT 2ýý2), DWV IGPGTCNN[ VJG[ CNN KPXQNXG J[DTKFKUCVKQP QH HNWQTGUEGPVN[ NCDGNNGF VCTIGVU U[PVJGUKUGF HTQO VJG O40# RQRWNCVKQP VQ CP CTTC[ EQPVCKPKPI û4( UGSWGPEGU HQT RQVGPVKCNN[ CNN û4(U KP C IGPQOG. 6QFC[, &0# CTTC[U CTG VJG EQOOQP VQQN HQT VTCPUETKRVQOG CPCN[UKU CPF VJG[ CTG WUGF KP OCP[ NCDQTCVQTKGU. 5GTKCN CPCN[UKU QH IGPG GZRTGUUKQP þ5#ô©) þíGNEWNGUEW GV CN. 199ó) ECP CNUQ DG WUGF HQT INQDCN VTCPUETKRVKQP CPCN[UKU þíGNEWNGUEW GV CN. 1997) CPF OC[ DG WUGF HQT KFGPVKHKECVKQP QH RTGXKQWUN[ WP-CPPQVCVGF IGPGU, UKPEG KV TGSWKTGU PQ RTKQT UGSWGPEG KPHQTOCVKQP DGHQTG KV ECP DG RGTHQTOGF. *QYGXGT, VJG OGVJQF KU SWKVG NCDQTKQWU þ*QTCM CPF 5P[FGT 2ýý2). &0# CTTC[U IKXG KPHQTOCVKQP QH YJKEJ IGPGU CTG GZRTGUUGF CPF VQ YJCV GZVGPV VJG[ CTG GZRTGUUGF. 6JWU, EJCPIGU KP VTCPUETKRVKQP HTQO C IKXGP VKOG QT EQPFKVKQP VQ CPQVJGT ECP DG SWCPVKHKGF CPF VJKU OC[ IKXG KPFKECVKQPU QH IGPGU HWPEVKQP CPF TGIWNCVKQP. ôGPGU VJCV RCTVKEKRCVG KP VJG UCOG EGNNWNCT RTQEGUUGU QHVGP UJCTG UKOKNCT VTCPUETKRVKQP RTQHKNGU CU KNNWUVTCVGF KP S. cerevisiae D[ G.I. VJG CPCN[UKU QH VJG OKVQVKE EGNN E[ENG þ%JQ GV CN. 1998), URQTWNCVKQP þ%JW GV CN. 1998), FKCWZKE UJKHV þ&G4KUK GV CN. 1997), OGQUKU þöTKOKI GV CN. 2ýýý), CPF XCTKQWU GPXKTQPOGPVCN VTCPUKVKQPU þôCUEJ GV CN. 2ýýýø ôCUEJ GV CN. 2ýý1). $[ OKPKPI GZRTGUUKQP FCVC, WUKPI UQHVYCTG CRRN[KPI ENWUVGTKPI CNIQTKVJOU VJCV ITQWR VQIGVJGT IGPGU YKVJ UKOKNCT VTCPUETKRVKQP RTQHKNGU, IGPGU YKVJ UKOKNCT GZRTGUUKQP ECP DG KFGPVKHKGF þ©KUGP GV CN. 1998ø 5JGTNQEM 2ýýý). (WPEVKQP QH QTRJCP IGPGU OC[ VJGP DG KPHGTTGF HTQO HWPEVKQP QH IGPGU VJCV JCXG C UKOKNCT VTCPUETKRVKQP RTQHKNG. 4GIWNCVQT[ KPHQTOCVKQP QDVCKPGF, WUKPI &0# CTTC[U KU LWUV CU XCNWCDNG. ôGPGU YKVJ UKOKNCT VTCPUETKRVKQP OKIJV CNUQ DG TGIWNCVGF D[ VJG UCOG OGEJCPKUOU, CPF VJGTGHQTG JCXG UKOKNCT TGIWNCVQT[ GNGOGPVU KP VJGKT RTQOQVGTU. öTQOQVGT CPCN[UKU QH UWEJ IGPGU ECP TGXGCN PGY TGIWNCVQT[ GNGOGPVU CPF KFGPVKH[ IGPGU VJCV CTG WPFGT TGIWNCVKQP QH C EGTVCKP VTCPUETKRVKQP HCEVQT. +FGMGT GV CN. þ2ýý1) CPCN[UGF VJG VTCPUETKRVQOG QH C PWODGT QH OWVCPVU YKVJ FGNGVGF ICNCEVQUG IGPGU CICKPUV C TGHGTGPEG UVTCKP CPF VJKU [KGNFGF XCNWCDNG KPHQTOCVKQP CDQWV VJG TGIWNCVQT[ PGVYQTM QH VJG ICNCEVQUG WVKNKUCVKQP RCVJYC[. öQVGPVKCN PGY ôCNü DKPFKPI UKVGU YGTG KFGPVKHKGF KP PKPG IGPGU D[ RTQOQVGT CPCN[UKU QH VJG IGPGU KP VJG UCOG GZRTGUUKQP ENWUVGTU CU VJG IGPGU YKVJ GUVCDNKUJGF ôCNü-DKPFKPI UKVGU. úCP[ OGVJQFU JCXG DGGP FGUETKDGF VQ KFGPVKH[ TGIWNCVQT[ GNGOGPVU KP RTQOQVGTU, G.I.: þ*WIJGU GV CN. 2ýýýø ÿGPUGP CPF ÷PWFUGP 2ýýýø ÷GNGU GV CN. 2ýý2ø XCP *GNFGP GV CN. 2ýýý). #P CNIQTKVJO PCOGF 4©&ñ%© JCU CNUQ DGGP FGXGNQRGF VJCV QPN[ TGSWKTGU C UKPING IGPQOG-YKFG UGV QH GZRTGUUKQP TCVKQU CPF VJG WRUVTGCO UGSWGPEG HQT GCEJ IGPG VQ QWVRWV UVCVKUVKECNN[ UKIPKHKECPV UGSWGPEG OQVKHU þ$WUUGOCMGT GV CN. 2ýý1). 6TCPUETKRVQOG CPCN[UKU ECP CNUQ DG WUGF HQT TGEQPUVTWEVKQP QH TGIWNCVQT[ RCVJYC[U. +P VJKU TGURGEV $C[GUKCP PGVYQTMU UGGO VQ DG C RQYGTHWN OGVJQF, CU KV ECP DG WUGF VQ KPHGT VJG UVCVKUVKECN
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ü3
FGRGPFGPEKGU DGVYGGP FKHHGTGPV IGPG RTQFWEVU DCUGF QP NCTIG UGVU QH VTCPUETKRVKQP RTQHKNGU HTQO FKHHGTGPV OWVCPVU þ(TKGFOCP GV CN. 2ýýýø 6QJ CPF *QTKOQVQ 2ýý2). 6JGTG CTG PQY PWOGTQWU GZCORNGU QH VJG WUG QH VTCPUETKRVQOG CPCN[UKU KP S. cerevisiae CPF QVJGT QTICPKUOU, CPF VTCPUETKRVKQP FCVC KU IGPGTCVGF KP UVCIIGTKPI COQWPVU. *QYGXGT, PQV OWEJ JCXG DGGP RWDNKUJGF VQ FCVG KP VJG HKGNF QH CRRNKGF [GCUV TGUGCTEJ DGUKFGU C HGY RCRGTU QP VTCPUETKRVQOG CPCN[UKU QH YKPG [GCUV þ$CEMJWU GV CN. 2ýý1ø %CXCNKGTK GV CN. 2ýýýø *CWUGT GV CN. 2ýý1). ùKVJ VJG UGSWGPEKPI QH QVJGT [GCUV URGEKGU, YG YKNN UQQP UGG VTCPUETKRVQOG CPCN[UKU QH QVJGT [GCUVU VJCP S. cerevisiae, G.I. YCU VJG IGPQOG-YKFG GZRTGUUKQP RCVVGTP QH S. pompe TGEGPVN[ KPXGUVKICVGF þúCVC GV CN. 2ýý2). (QT OQTG GZVGPUKXG TGXKGYU QH GZCORNGU CPF VJG VGEJPQNQI[ UGG G.I.: þ*QTCM CPF 5P[FGT 2ýý2ø ÷PWFUGP 2ýý2ø ÷WOCT CPF 5P[FGT 2ýý1ø öoTG\-ûTVsP GV CN. 2ýý2). 11.4.3 )NQDCN OWVCPV CPCN[UKU ôNQDCN OWVCPV CPCN[UKU RQVGPVKCNN[ CNNQYU WU VQ VGUV VJG HWPEVKQPCN TQNG QH CNN IGPGU FKTGEVN[ CPF UKOWNVCPGQWUN[ D[ GZCOKPKPI VJG RJGPQV[RG QH CNN OWVCPVU. 4GEGPVN[ EQPUVTWEVKQP QH C PGCTN[ EQORNGVG EQNNGEVKQP QH IGPG-FKUTWRVKQP OWVCPVU þ9õî QH CPPQVCVGF û4(U) KP S. cerevisiae YCU HKPKUJGF CPF RWDNKUJGF þôKCGXGT GV CN. 2ýý2). 6JKU KORTGUUKXG EQNNGEVKQP YCU EQPUVTWEVGF D[ C ÷CPúð FGNGVKQP ECUUGVVG YJKEJ KP GCEJ TGUWNVKPI OWVCPV EQPVCKPGF VYQ FKUVKPEV 2ý PWENGQVKFG UGSWGPEGU VJCV UGTXG CU ÂOQNGEWNCT DCTEQFGU• VJCV CTG WPKSWG HQT GCEJ OWVCPV þ5JQGOCMGT GV CN. 199õø ùKP\GNGT GV CN. 1999). 6JG OQNGEWNCT DCTEQFGU CNNQY RCTCNNGN CPCN[UKU QH VJG RJGPQV[RGU QH CNN FGNGVKQP OWVCPVU VQ DG RGTHQTOGF WUKPI EQORGVKVKXG ITQYVJ CUUC[U. +P VJGUG CUUC[U, VJG HKVPGUU EQPVTKDWVKQP QH GCEJ IGPG KU CUUGUUGF SWCPVKVCVKXGN[ D[ J[DTKFKUCVKQP QH VJG OQNGEWNCT DCTEQFGU VQ &0# CTTC[U QH VJG EQORNGOGPVCT[ DCTEQFG UGSWGPEGU CHVGT DGKPI ö%4-CORNKHKGF HTQO IGPQOKE &0#. ôKCXGT GV CN. þ2ýý2) RGTHQTOGF EQORGVKVKXG ITQYVJ CUUC[U WPFGT UKZ YGNN-UVWFKGF EQPFKVKQPU CPF KFGPVKHKGF PGY IGPGU VQ DG PGEGUUCT[ HQT QRVKOCN ITQYVJ WPFGT VJG VGUVGF EQPFKVKQPU. $KTTGNN GV CN. þ2ýý2) WUGF VJG EQNNGEVKQP VQ VGUV TGURQPUG VQ &0#-FCOCIKPI CIGPVU, CPF RTGXKQWU VQ VJG EQORNGVKQP QH VJG IGPG-FKUTWRVKQP EQNNGEVKQP VJGTG YGTG CNUQ UQOG UVWFKGU YKVJ KPEQORNGVG EQNNGEVKQPU QH IGPG-FKUTWRVKQP OWVCPVU þ$GPPGVV GV CN. 2ýý1ø $KTTGNN GV CN. 2ýý1ø ùKP\GNGT GV CN. 1999). ôGPG-FKUTWRVKQP EQNNGEVKQPU QHHGT JWIG RQUUKDKNKVKGU, DWV HWTVJGT FGXGNQROGPVU CTG PGEGUUCT[. 6JG HCKNWTG QH CP[ IKXGP OWVCPV VQ TGXGCN C RJGPQV[RG OC[ DG VJG EQPUGSWGPEG QH GKVJGT IGPGVKE TGFWPFCPE[ QT QH GHHGEVKXG JQOGQUVCVKE EQPVTQNU YKVJKP VJG EGNN þ$TGPV 2ýýýø &GNPGTK GV CN. 2ýý1). úCMKPI FQWDNG þQT OQTG) MPQEMQWV EQODKPCVKQPU QH VJG CRRTQZKOCVGN[ ü8ýý PQPGUUGPVKCN IGPGU KP S. cerevisiae OKIJV TGXGCN CPF ENCTKH[ VJG HWPEVKQP QH UQOG QH VJGUG IGPGU. # OGVJQF ECNNGF U[PVJGVKE IGPGVKE CTTC[ þ5ô#) CPCN[UKU JCU DGGP FGXGNQRGF HQT VJKU RWTRQUG þ6QPI GV CN. 2ýý1). *QYGXGT, LWUV OCMKPI GXGT[ FQWDNG MPQEMQWV EQODKPCVKQP QH VJG PQPGUUGPVKCN IGPGU YQWNF [KGNF 23 OKNNKQP OWVCPVU þìWG CPF ùKP\GNGT 2ýý2). (WTVJGT IGPGVKE OCPKRWNCVKQP QP VJG OWVCPV EQNNGEVKQP KU CNUQ RQUUKDNG D[ VTCPUHQTOKPI VJG GPVKTG RQQN YKVJ CP CRRTQRTKCVG RNCUOKF QT TGRQTVGT þûQK GV CN. 2ýý1ø ìWG CPF ùKP\GNGT 2ýý2). úGVJQFU HQT UKOWNVCPGQWU CPCN[UKU QH QXGT-GZRTGUUKQP QH IGPGU
3üü
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
QP C IGPQOG-YKFG DCUKU CTG CNUQ PGGFGF. 5WEJ C OGVJQF JCU DGGP ETGCVGF KP Escherichia coli D[ VTCPUHQTOKPI C UVTCKP YKVJ C IGPQOKE NKDTCT[, GPTKEJKPI VJG EGNN RQRWNCVKQP KP VTCPUHQTOCPVU EQPVCKPKPI VJG VTCKV EQPHGTTGF IGPGþU), CPF HKPCNN[ WUKPI &0# CTTC[U VQ KFGPVKH[ GPTKEJGF IGPG KPUGTVU þôKNN GV CN. 2ýý2). ûVJGTU JCXG VCMGP CPQVJGT CRRTQCEJ HQT IGPGTCVKPI NCTIG EQNNGEVKQPU QH FGHKPGF [GCUV OWVCPVU. 6JG[ JCXG WUGF C TCPFQO VTCPURQUQP OWVCIGPGUKU UVTCVGI[ VJCV GZRNQKVU VTCPURQUQPU CDKNKV[ VQ ECTT[ XCTKQWU TGRQTVGT IGPGU, GRKVQRG VCIU, CPF TGIWNCVQT[ GNGOGPVU þ$WTPU GV CN. 199üø 4QUU-úCEFQPCNF GV CN. 1997ø 4QUU-úCEFQPCNF GV CN. 1999ø 5OKVJ GV CN. 199ó). 6JKU UVTCVGI[ IKXGU VJG RQUUKDKNKV[ VQ FGVGEV VJG GZRTGUUKQP NGXGN KP XKXQ, ENCTKH[ VJG UWDEGNNWNCT NQECNKUCVKQP, CPF UETGGP HQT RJGPQV[RGU UWEJ CU ITQYVJ, VGORGTCVWTG UGPUKVKXKV[, CPF EGNNWNCT TGURQPUGU VQ EJGOKECNU þ6CMCJCUJK GV CN. 2ýý1). 6JG VTCPURQUQP UVTCVGI[ KU CNUQ OQTG CRRNKECDNG VQ QTICPKUOU QVJGT VJCP S. cerevisiae YJGTG NQPIGT UVTGVEJGU QH JQOQNQI[ CTG PGGFGF HQT TGEQODKPCVKQP VQ QEEWT þìWG CPF ùKP\GNGT 2ýý2). S. pombe JCU DGGP UWDLGEV VQ TCPFQO KPUGTVKQPCN IGPQOG-YKFG OWVCIGPGUKU þ%JWC GV CN. 2ýýýø *QHHOCP CPF ùGNVQP 2ýýý). *QYGXGT, FWG VQ VJG PQV EQORNGVGN[ TCPFQOKUGF PCVWTG QH VTCPURQUQP KPUGTVKQP KV KU XGT[ FKHHKEWNV VQ OWVCIGPKUG CNN IGPGU YKVJKP C IGPQOG þ%JWC GV CN. 2ýýýø *QHHOCP CPF ùGNVQP 2ýýýø ÷WOCT CPF 5P[FGT 2ýý1). 11.4.4 ÿTQVGQOG CPCN[UKU öTQVGQOKEU, VJG UKOWNVCPGQWU NCTIG-UECNG CPCN[UKU QH VJG RTQVGKP EQORQPGPV QH CP QTICPKUO þöCPFG[ CPF úCPP 2ýýý), RQVGPVKCNN[ DTKPIU WU QPG NGXGN ENQUGT VQ WPFGTUVCPFKPI IGPG HWPEVKQP VJCP VTCPUETKRVQOG CPCN[UKU, DWV VJG NGXGN QH FKHHKEWNV[ KU CNUQ JKIJGT. öTQVGQOG CPCN[UKU EQXGTU OCP[ CURGEVU QH RTQVGKP EJCTCEVGTKUCVKQP UWEJ CU KFGPVKHKECVKQP, SWCPVKHKECVKQP, EJCTCEVGTKUCVKQP CPF KFGPVKHKECVKQP QH RQUVVTCPUNCVKQPCN OQFKHKECVKQPU, CPF EGNNWNCT NQECNKUCVKQP. 6YQ-FKOGPUKQPCN þ2&) IGN GNGEVTQRJQTGUKU JCU DGGP WUGF HQT CPCN[UKU QH VJG RTQVGQOG UKPEG VJG NCVG 197ýU CPF KU UVKNN CP KORQTVCPV VGEJPKSWG HQT UGRCTCVKQP CPF SWCPVKHKECVKQP KP RTQVGQOKEU VQFC[ þ(WVEJGT GV CN. 1999ø öGTTQV GV CN. 1999). 6JG RTQVGKPU TGUQNXGF D[ 2& GNGEVTQRJQTGUKU ECP DG KFGPVKHKGF CPF CPCN[UGF HQT G.I. RQUV-VTCPUNCVKQPCN OQFKHKECVKQPU D[ VJG WUG QH VJG OQUV KORQTVCPV VGEJPKSWG KP RTQVGQOKEU VQFC[, PCOGN[, OCUU URGEVTQUEQR[ þV[RKECNN[ ú#ï&+-6û(-ú5)þ#GDGTUQNF CPF ôQQFNGVV 2ýý1ø $NCEMUVQEM CPF úCPP 2ýýýø úCPP GV CN. 2ýý1) EQODKPGF YKVJ IGPQOKE KPHQTOCVKQP. *QYGXGT, URGEKHKE ENCUUGU QH RTQVGKPU CPF NQY CDWPFCPEG RTQVGKPU CTG MPQYP VQ DG CDUGPV QT WPFGTTGRTGUGPVGF KP 2& IGN RCVVGTPU þôQQFNGVV CPF ;K 2ýý2), CPF FWG VQ VJKU CPF QVJGT KUUWGU þUGG G.I.: þôQQFNGVV CPF ;K 2ýý2ø ûNKXGT GV CN. 2ýý2)) PGY OGVJQFU HQT CPCN[UKU QH RTQVGKPU KP EQORNGZ OKZVWTGU CXQKFKPI VJG WUG QH 2& IGNU CTG WPFGT FGXGNQROGPV þôQQFNGVV CPF ;K 2ýý2). 6JG HQEWU KP TGEGPV GHHQTVU KP RTQVGQOKEU JCU UJKHVGF VQ SWCPVKVCVKXG OGVJQFU VJCV CNNQY RTQVGKP NGXGNU VQ DG OGCUWTGF INQDCNN[. ôNQDCN CPCN[UKU OGVJQFU KP RTQVGQOKEU CTG UVKNN WPFGT FGXGNQROGPV CPF CTG CNUQ UVCTVKPI VQ CRRGCT KP HKGNFU HQT CPCN[UKU QH RTQVGKP DKQEJGOKUVT[ CPF EGNNWNCT NQECNKUCVKQP, YJKEJ EQPVTKDWVGU YKVJ KORQTVCPV KPHQTOCVKQP CDQWV RTQVGKP HWPEVKQP CPF TGIWNCVKQP. 4GEGPVN[ VTCPURQUQPU YGTG CRRNKGF HQT VCIIKPI CPF KPFKTGEV KOOWPQHNWQTGUEGPEG VQ FQ C NCTIG-UECNG KPXGUVKICVKQP CUUKIPKPI C UWDEGNNWNCT
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3üó
NQECNKUCVKQP HQT CDQWV JCNH VJG S. cerevisiae RTQVGKPU þ÷WOCT GV CN. 2ýý2). öTQVGQOG EJKRU CTG DGKPI FGXGNQRGF HQT INQDCN CPCN[UKU QH RTQVGKP DKQEJGOKUVT[ þúCE$GCVJ CPF 5EJTGKDGT 2ýýýø <JW GV CN. 2ýýýø <JW GV CN. 2ýý1) CPF ECP DG WUGF HQT UVWF[KPI RTQVGKP KPVGTCEVKQPU, UWTXG[KPI RQUV-VTCPUNCVKQPCN OQFKHKECVKQPU CPF KFGPVKH[KPI FQYPUVTGCO VCTIGVU QH FKHHGTGPV ENCUUGU QH GP\[OGU, UWEJ CU MKPCUGU, RJQURJCVCUGU CPF WDKSWKVKPU þ<JW CPF 5P[FGT 2ýý1ø <JW CPF 5P[FGT 2ýý2). +PXGUVKICVKQPU QH VJG EQTTGNCVKQP DGVYGGP O40# CPF RTQVGKP GZRTGUUKQP KP S. cerevisiae JCXG UJQYGF VJG FKHHKEWNVKGU QH RTGFKEVKPI VJG [GCUV RTQVGQOG FKTGEVN[ HTQO VJG VTCPUETKRVQOG þô[IK GV CN. 1999ø +FGMGT GV CN. 2ýý1). 6JG EQTTGNCVKQP DGVYGGP O40# CPF RTQVGKP CDWPFCPEG KU XGT[ RQQT HQT UQOG IGPGU, UWIIGUVKPI VJCV RQUV-VTCPUETKRVKQPCN TGIWNCVKQP RNC[U CP KORQTVCPV TQNG. ö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þ(KGNFU CPF 5QPI 1989) CPF TGEGPVN[, KV JCU DGGP WUGF HQT IGPQOG-YKFG UVWFKGU þ+VQ GV CN. 2ýýýø ñGV\ GV CN. 2ýýý). 6YQ-J[DTKF CUUC[U CTG GCU[ VQ FQ CPF TGCFKN[ CRRNKECDNG VQ QVJGT GWMCT[QVKE U[UVGOU, DWV PQV CNN RTQVGKPU ECP DG CPCN[UGF HQT VJGKT KPVGTCEVKQPU D[ VJG CUUC[, YJKEJ CNUQ RTQFWEGU OCP[ HCNUG-RQUKVKXG QWVEQOGU. *QYGXGT, HWVWTG VYQ-J[DTKF VGEJPQNQIKGU OKIJV QXGTEQOG UQOG QH VJG RTGUGPV-FC[ NKOKVCVKQPU þ÷WOCT CPF 5P[FGT 2ýý1), CPF RTQVGKP-RTQVGKP KPVGTCEVKQPU OKIJV CNUQ DG CPCN[UGF YKVJ RTQVGKP EJKRU þúCE$GCVJ CPF 5EJTGKDGT 2ýýýø <JW GV CN. 2ýýýø <JW GV CN. 2ýý1). 4GEGPVN[, VJG VYQ HKTUV NCTIG-UECNG CHHKPKV[ VCIIKPI CPF OCUU URGEVTQUEQR[ CPCN[UKU QH RTQVGKP EQORNGZGU KP [GCUV YGTG RWDNKUJGF þôCXKP GV CN. 2ýý2ø *Q GV CN. 2ýý2). 6JGUG UVWFKGU KFGPVKHKGF JWPFTGFU QH PQXGN RTQVGKP-RTQVGKP KPVGTCEVKQPU CPF RTQVGKP EQORNGZGU CPF RTQXKFG KPUKIJV KPVQ VJG OGEJCPKUOU D[ YJKEJ RTQVGKP EQORNGZGU CTG EQQTFKPCVGF KPVQ JKIJ-QTFGT PGVYQTMU TGURQPUKDNG HQT OCP[ HWPFCOGPVCN EGNNWNCT RTQEGUUGU þ<JW CPF 5P[FGT 2ýý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þ+[GT GV CN. 2ýý1ø 4GP GV CN. 2ýýý). 4GEGPVN[, C NCTIG TGIWNCVQT[ PGVYQTM UVTWEVWTG YCU TGEQPUVTWEVGF HQT S. cerevisiae DCUGF QP VJG KPVGTCEVKQPU HQWPF DGVYGGP OQUV QH VJG VTCPUETKRVKQPCN TGIWNCVQTU GP-
3üõ
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
EQFGF KP S. cerevisiae CPF VJG IGPGU CETQUU VJG IGPQOG þïGG GV CN. 2ýý2). #NUQ RTQVGKP EJKRU JCXG RQVGPVKCN HQT UVWF[KPI RTQVGKP-&0# KPVGTCEVKQPU þ<JW CPF 5P[FGT 2ýý2). 11.4.þ /GVCDQNQOG CPCN[UKU 6JG OGVCDQNQOG, EQPUKUVKPI QH CNN VJG KPVTCEGNNWNCT OGVCDQNKVGU, KU C HWPEVKQP QH VJG HNWZQOG, YJKEJ CICKP KU C HWPEVKQP QH VJG QVJGT ÂQOGU• CU OGPVKQPGF GCTNKGT. 6JWU, VJG OGVCDQNQOG TGRTGUGPVU KPVGITCVKXG KPHQTOCVKQP, CPF VJGTGHQTG, CPCN[UKU QH VJG OGVCDQNQOG OC[ IKXG HWTVJGT KPUKIJV KPVQ VJG HWPEVKQP QH IGPGU þ({TUVGT GV CN. 2ýý2). 6JG TGEGPV CFXCPEGU KP HWPEVKQPCN IGPQOKEU JCXG WTIGF VJG PGGF HQT INQDCN CPCN[UKU QH VJG OGVCDQNQOG þ(KGJP 2ýý1ø (KGJP 2ýý2). ùKVJ VJG FGXGNQROGPV QH CPCN[VKECN VGEJPQNQIKGU HQT UGRCTCVKQP, SWCPVKHKECVKQP, CPF KFGPVKHKECVKQP KV KU VQFC[ RQUUKDNG VQ OGCUWTG 1óý KPVTCEGNNWNCT OGVCDQNKVGU D[ WUKPI ô%-ú5 þ4QGUUPGT GV CN. 2ýýý). 6JG OGVCDQNKUO QH S. cerevisiae KU GUVKOCVGF VQ KPXQNXG CRRTQZKOCVGN[ óõý NQY OQNGEWNCT OGVCDQNKVGU þûNKXGT GV CN. 1998), CPF KP C TGEGPVN[ TGEQPUVTWEVGF OGVCDQNKE PGVYQTM HQT S. cerevisiae CDQWV õýý OGVCDQNKVGU YGTG KFGPVKHKGF þ({TUVGT GV CN. 2ýý3). ùKVJ VJKU TGNCVKXGN[ NQY PWODGT QH OGVCDQNKVGU KV OC[ KP VJG PGCT HWVWTG DG RQUUKDNG VQ OGCUWTG VJG HWNN OGVCDQNQOG QH S. cerevisiae. 6JG HKGNF KU UVKNN KP KVU KPHCPE[ CPF UQ HCT QPN[ C HGY TGUGCTEJ RCRGTU JCXG DGGP RWDNKUJGF. *QYGXGT, VJG RQYGT QH OGVCDQNQOKEU JCU CNTGCF[ DGGP KNNWUVTCVGF D[ C UVWF[ QP S. cerevisiae UJQYKPI VJCV OWVCVKQPU YKVJ UKNGPV RJGPQV[RGU ECP DG TGXGCNGF D[ EJCPIGU KP OGVCDQNKVG RTQHKNGU þ4CCOUFQPM GV CN. 2ýý1). 4CCOUFQPM GV CN. þ2ýý1) CNUQ UJQYGF VJCV FGNGVKQP OWVCPVU JCXKPI NQUV UKOKNCT HWPEVKQPU JCXG UKOKNCT OGVCDQNKVG RTQHKNGU. 6JG[ RTQRQUG VJCV VJKU ECP DG WUGF HQT CUUKIPKPI HWPEVKQP VQ QTRJCP IGPGU D[ EQORCTKPI VJG EJCPIGU KP OGVCDQNKVG RTQHKNG VJCV KU RTQFWEGF D[ FGNGVKPI VJG IGPG YKVJ C NKDTCT[ QH RTQHKNGU IGPGTCVGF D[ KPFKXKFWCNN[ FGNGVKPI IGPGU QH MPQYP HWPEVKQP. úGVCDQNQOG CPCN[UKU KU CNUQ WUGHWN HQT CPCN[UKU QH YKPG CPF DTGYGTU [GCUVU. 6JKU JCU CNTGCF[ DGGP GZGORNKHKGF D[ þ©INKPVQP GV CN. 2ýý2), YJQ WUGF OGVCDQNKVG RTQHKNKPI VQ UJQY VJG GHHGEV QH IGPGVKE GPIKPGGTKPI QH S. cerevisiae QP OGVCDQNKVGU VJCV CTG KORQTVCPV VQ VJG HNCXQWT QH YKPG. 11.4.7 (NWZQOG CPCN[UKU #PCN[UKU QH VJG HNWZQOG þCNN HNWZGU KP VJG EGNN) IKXGU KPHQTOCVKQP CDQWV YJKEJ RCVJYC[U CTG CEVKXG CPF VQ YJCV GZVGPV. 6JGTGD[, C DGVVGT WPFGTUVCPFKPI QH YJCV KU QEEWTTKPI KPUKFG VJG EGNN CV C IKXGP VKOG KU QDVCKPGF. úQUV HNWZGU ECPPQV DG OGCUWTGF FKTGEVN[ KP XKXQ, DWV VJTQWIJ VJG WUG QH 13%-NCDGNNGF UWDUVTCVGU HQNNQYGF D[ CPCN[UKU QH VJG NCDGNNKPI RCVVGTP QH KPVTCEGNNWNCT OGVCDQNKVGU CP GUVKOCVKQP QH VJG HNWZGU ECP DG QDVCKPGF XKC C OCVJGOCVKECN OQFGN QH VJG OGVCDQNKUO. 6JG NCDGNNKPI RCVVGTP QH 13% KP KPVTCEGNNWNCT OGVCDQNKVGU OC[ DG CPCN[UGF WUKPI GKVJGT ô%-ú5 þ%JTKUVGPUGP CPF 0KGNUGP 2ýýý) QT 0ú4 þ5\[RGTUMK 1998ø ùGPFKUEJ GV CN. 1997), CPF VJG NCDGNNKPI RCVVGTP JCU DGGP RTQXGF XGT[ WUGHWN HQT KFGPVKHKECVKQP QH VJG OGVCDQNKE PGVYQTM UVTWEVWTG þ%JTKUVGPUGP CPF 0KGNUGP 2ýýý). (TQO VJG NCDGNNKPI
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ü7
RCVVGTP CPF OGVCDQNKVG DCNCPEKPI C FGVCKNGF CPCN[UKU VJCV KPENWFGU HNWZ GUVKOCVKQP, KFGPVKHKECVKQP QH VJG PGVYQTM VQRQNQI[, OGVCDQNKE EJCPPGNNKPI, CPF UWDEGNNWNCT EQORCTVOGPVCVKQP ECP DG QDVCKPGF þ%JTKUVGPUGP CPF 0KGNUGP 2ýýýø ùKGEJGTV 2ýý1). 6JGTG CTG UGXGTCN GZCORNGU QH VJKU OGVCDQNKE PGVYQTM CPCN[UKU CRRTQCEJ KP [GCUV HQT GUVKOCVKPI VJG HNWZ FKUVTKDWVKQPU QH VJG EGPVTCN ECTDQP OGVCDQNKUO. $QVJ KP S. cerevisiae, YJGTG VJG HNWZGU YGTG GUVKOCVGF CV FKHHGTGPV EQPFKVKQPU QH INWEQUG TGRTGUUKQP þôQODGTV GV CN. 2ýý1) CPF KP S. kluyveri YJGTG KV YCU GUVKOCVGF HQT TGURKTQ-HGTOGPVCVKXG CPF TGURKTCVQT[ ITQYVJ þúòNNGT GV CN. 2ýý2).
11.5 (WPEVKQPCN IGPQOKEU KP OGVCDQNKE GPIKPGGTKPI +P OGVCDQNKE GPIKPGGTKPI, VJG IQCN KU VQ NKPM IGPGU CPF RCVJYC[U VQ RJGPQV[RG. ñPVKN PQY, VJKU JCU DGGP FQPG D[ UGCTEJKPI HQT EJCPIGU KP VJG EGNNWNCT RJGPQV[RG VJCV CTG CUUQEKCVGF YKVJ OQFKHKECVKQPU KPVTQFWEGF KPVQ C RCTVKEWNCT IGPG. $[ CFCRVKPI OGVJQFU HTQO HWPEVKQPCN IGPQOKEU KV KU PQY RQUUKDNG VQ CRRN[ CP KPXGTUG CRRTQCEJ CPF KFGPVKH[ VJG IGPGþU) QT GPXKTQPOGPVCN EQPFKVKQPþU) IGPGTCVKPI C FGUKTCDNG RJGPQV[RG. (WPEVKQPCN IGPQOKEU RTQXKFGU PGY VQQNU CPF CRRTQCEJGU HQT WPFGTUVCPFKPI, OQFGNNKPI, CPF OCPKRWNCVKPI EGNNU CPF JGTGD[ CNNQYU VJG CRRNKECVKQP QH C IGPQOG-YKFG CRRTQCEJ. ©XGP VJQWIJ, VJG HWPEVKQP QH VYQ VJKTFU QH VJG S. cerevisiae IGPGU CTG MPQYP QP VJG DKQEJGOKECN NGXGN, VJG HWPEVKQP QH OCP[ QH VJGUG IGPGU CV VJG RJGPQV[RKE NGXGN KU WPMPQYP. 6JGTGHQTG, VJG OGVCDQNKE GPIKPGGTKPI IQCN QH KFGPVKH[KPI IGPGU VJCV EQPHGT C RCTVKEWNCT RJGPQV[RG KU EQPEGRVWCNN[ CPF OGVJQFQNQIKECNN[ EQPITWGPV YKVJ EGPVTCN KUUWGU KP HWPEVKQPCN IGPQOKEU þ$CKNG[, 1999). 5KPEG HWPEVKQPCN IGPQOKEU YKNN PQV QPN[ GNWEKFCVG YJCV VJG IGPGU FQ, DWV YKNN CNUQ JGNR FGVGTOKPG YJGP, YJGTG CPF JQY VJG[ CTG GZRTGUUGF CU CP QTICPKUGF U[UVGO. *GPEG, HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI YKNN EQPVTKDWVG VQ VJG UCOG IQCN PCOGN[ QDVCKPKPI CP KORTQXGF WPFGTUVCPFKPI QH EGNNWNCT HWPEVKQP CV VJG YJQNG-EGNN NGXGN. 6JG VQQNU QH HWPEVKQPCN IGPQOKEU ECP CNUQ DG CRRNKGF FKTGEVN[ KP OGVCDQNKE GPIKPGGTKPI VQ HKPF VCTIGVU HQT KORTQXGF RJGPQV[RG. 6TCPUETKRVQOG CPCN[UKU YKVJ VJG WUG QH &0# CTTC[U JCU OCP[ WUGHWN CRRNKECVKQPU KP VJG CPCN[UKU RCTV QH OGVCDQNKE GPIKPGGTKPI. 6JG OQUV FKTGEV CRRNKECVKQP QH &0# CTTC[U VQ OGVCDQNKE GPIKPGGTKPI YKNN DG VJG KFGPVKHKECVKQP QH FKUETKOKPCVQT[ IGPGU EJCTCEVGTKUVKE QH FGUKTGF RJ[UKQNQIKECN UVCVGU, UWEJ CU VJQUG EQPVTKDWVKPI VQ JKIJ RTQFWEVKXKV[ þ5VCHHQTF CPF 5VGRJCPQRQWNQU 2ýý1). $[ EQORCTKPI VTCPUETKRVKQP RTQHKNGU QH C UVTCKP KORTQXGF UWEEGUUHWNN[ D[ OGVCDQNKE GPIKPGGTKPI YKVJ VJG TGHGTGPEG UVTCKP CNN RQUUKDNG RNGQVTQRJKE GHHGEVU ECP DG OCRRGF. 6JG CPCN[UKU OKIJV RQKPV VQ IGPGU, YJQUG EJCPIGU KP GZRTGUUKQP KPHNWGPEGU VJG RJGPQV[RG RQUKVKXGN[ QT PGICVKXGN[, CPF JGTGD[, KV OC[ RQKPV VQ PGY VCTIGVU HQT C PGZV TQWPF QH OGVCDQNKE GPIKPGGTKPI. 6JKU OC[ NGCF VQ KFGPVKHKECVKQP QH VCTIGVU VJCV YQWNF JCXG DGGP FKHHKEWNV VQ CEJKGXG YKVJQWV C YJQNG-EGNN CRRTQCEJ. *QYGXGT, CPCN[UKPI LWUV QPG JKIJ-RTQFWEKPI UVTCKP OKIJV OCMG KV FKHHKEWNV VQ KFGPVKH[ MG[ IGPGU HQT GPJCPEKPI VJG RTQFWEVKXKV[ QT [KGNF. $[ CPCN[UKPI VYQ QT OQTG
3ü8
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
+FGPVKH[ ECPFKFCVG IGPGU HQT GPJCPEKPI RTQFWEVKXKV[ QT [KGNF
2TQFWEVKXKV[/[KGNF
MCPKRWNCVG ECPFKFCVG IGPG(U)
(KI. 5. /GVJQF VQ KFGPVKH[ VCTIGVU HQT OGVCDQNKE GPIKPGGTKPI HTQO IGPQOG-YKFG VTCPUETKRVKQP CPCN[UKU. # PWODGT QH UVTCKPU YKVJ FKHHGTGPV RTQFWEVKXKV[/[KGNF/UWDUVTCVG WRVCMG KU CPCN[UGF VQ KFGPVKH[ FKHHGTGPVN[ GZRTGUUGF IGPGU, YJKEJ OKIJV DG MG[ GPJCPEGTU HQT VJG FGUKTGF RCTCOGVGT. 6JG ECPFKFCVG IGPGU CTG FGNGVGF QT QXGT-GZRTGUUGF KP C PGY E[ENG QH OGVCDQNKE GPIKPGGTKPI.
JKIJ RTQFWEKPI UVTCKPU YKVJ FKHHGTGPV RTQFWEVKXKV[ QT [KGNF, KFGPVKHKECVKQP KU OWEJ OQTG NKMGN[ D[ RKEMKPI WR IGPGU YKVJ UKIPKHKECPV EJCPIGU JCXKPI VJG UCOG FKTGEVKQP KP CNN CPCN[UGF UVTCKPU þUGG (KIWTG ó). +P QWT NCDQTCVQT[, YG GZRNQKVGF VJKU CRRTQCEJ D[ WUKPI &0# CTTC[U VQ CPCN[UG S. cerevisiae UVTCKPU YKVJ FKHHGTGPV FGITGGU QH KPETGCUGF HNWZ VJTQWIJ VJG ICNCEVQUG WVKNKUCVKQP RCVJYC[. 6JG UVTCKPU CPCN[UGF JCF RTGXKQWUN[ DGGP TCVKQPCNN[ FGUKIPGF CPF KORTQXGF D[ OGVCDQNKE GPIKPGGTKPI QH VJG TGIWNCVQT[ RCVJYC[ þûUVGTICCTF GV CN. 2ýýýCø UGG (KIWTG 3). 6JTQWIJ IGPQOG-YKFG GZRTGUUKQP CPCN[UKU WUKPI &0# CTTC[U QH VJG YKNF V[RG CPF VYQ GPIKPGGTGF UVTCKPU, YG KFGPVKHKGF C PGY VCTIGV HQT OGVCDQNKE GPIKPGGTKPI. 6JKU VCTIGV YCU RTQXGP VQ DG C MG[ IGPG KP GPJCPEKPI VJG HNWZ VJTQWIJ VJG ICNCEVQUG WVKNKUCVKQP RCVJYC[, CPF QXGT-GZRTGUUKPI VJG IGPG GPJCPEGF VJG ICNCEVQUG HNWZ YKVJ 7ýî þWPRWDNKUJGF TGUWNVU). &0# CTTC[U CTG CNUQ CRRNKECDNG HQT KPXGTUG OGVCDQNKE GPIKPGGTKPI QH YJKEJ VJG EQPEGRV KU VQ HKTUV, KFGPVKH[, EQPUVTWEV, QT ECNEWNCVG C FGUKTGF RJGPQV[RG, VJGP FGVGTOKPG VJG IGPGVKE HCEVQTU EQPHGTTKPI VJCV RJGPQV[RG CPF HKPCNN[ GPFQYKPI VJCV RJGPQV[RG QP CPQVJGT UVTCKP QT QTICPKUO D[ FKTGEVGF IGPGVKE OCPKRWNCVKQP þ$CKNG[ GV CN. 199õ). úCRRKPI QH OWVCVKQPU OC[ DG FQPG VJTQWIJ EQORCTKUQP QH UGSWGPEG QT VTCPUETKRVKQP RTQHKNGU HTQO UVTCKPU YKVJ VJG FGUKTGF RJGPQV[RG UWEJ CU ENCUUKECNN[
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ü9
KORTQXGF/JKIJ RTQFWEKPI UVTCKPU YKVJ EQTTGURQPFKPI YKNF V[RG UVTCKPU þUGG (KIWTG õ). 6JWU, IGPGU, YJKEJ CTG FKHHGTGPVN[ GZRTGUUGF KP VJG JKIJ-RTQFWEKPI UVTCKP, CTG KFGPVKHKGF. ûPG, UGXGTCN, QT C EQODKPCVKQP QH VJGUG IGPGU OC[ DG ECWUKPI VJG FGUKTGF RJGPQV[RG. 5WEJ C UVTCVGI[ QH KPXGTUG OGVCDQNKE GPIKPGGTKPI OC[ DG WUGF VQ KFGPVKH[ PQXGN VCTIGVU HQT OGVCDQNKE GPIKPGGTKPI, CPF VJKU JCU DGGP VGUVGF TGEGPVN[ QP Z[NQUG-WVKNKUKPI S. cerevisiae UVTCKPU þùCJNDQO GV CN. 2ýý3)þUGG (KIWTG 2). 6JG ENCUUKECNN[ KORTQXGF UVTCKP, YJKEJ YCU EQORCTGF VQ KVU TGHGTGPEG UVTCKP YCU IGPGTCVGF D[ EJGOKECN OWVCIGPGUKU QH C Z[NQUG WVKNKUKPI S. cerevisiae UVTCKP CPF UGNGEVGF HQT JKIJ ITQYVJ TCVG QP Z[NQUG þùCJNDQO GV CN. 2ýý3). 6JG VYQ UVTCKPU YGTG ITQYP KP EJGOQUVCV EWNVKXCVKQPU QP INWEQUG, INWEQUG ë Z[NQUG YJKNG VJG KORTQXGF UVTCKP YCU ITQYP QP Z[NQUG CNQPG. 6JG VTCPUETKRVQOG QH VJG VYQ UVTCKPU WPFGT VJG FKHHGTGPV EQPFKVKQPU YGTG CPCN[UGF YKVJ &0# CTTC[U, CPF C PWODGT QH IGPGU, DGNKGXGF VQ DG KPVGTGUVKPI HQT VJG Z[NQUG OGVCDQNKUO, YGTG HQWPF VQ JCXG CNVGTGF GZRTGUUKQP KP VJG ENCUUKECNN[ KORTQXGF UVTCKP þùCJNDQO GV CN. 2ýý3). ûPG QH VJG IGPGU, YJKEJ CNVGTGF GZRTGUUKQP, GPEQFGU C VTCPUETKRVKQPCN TGIWNCVQT þöGV18), YJKEJ UWDUGSWGPVN[ YCU OCPKRWNCVGF KP VJG TGHGTGPEG UVTCKP. *QYGXGT, PGKVJGT QXGT-GZRTGUUKQP PQT FGNGVKQP CHHGEVGF VJG ITQYVJ QP Z[NQUG þùCJNDQO GV CN. 2ýý3). 6JKU UJQYU VJG FKHHKEWNVKGU QH UGNGEVKPI VJG TKIJV IGPGþU) VQ VCTIGV HTQO VJG CPCN[UKU, UKPEG KV OKIJV DG CP[ QH VJG IGPGU YKVJ EJCPIGF GZRTGUUKQP QT C EQODKPCVKQP. 5WEEGUUHWN VTCPUHGT QH OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU HTQO NCDQTCVQT[ UVTCKPU VQ RTQFWEVKQP UVTCKPU ECP DG FKHHKEWNV FWG VQ IGPGVKE FKHHGTGPEGU. ôGPGVKE GPIKPGGTKPI KP KPFWUVTKCN [GCUV OCKPN[ KPXQNXGU IGPGVKE YQTM QP JCRNQKFU CPF FKRNQKFU HQNNQYGF D[ ETQUUKPI. &WTKPI ETQUUKPI, RQQTN[ FGHKPGF IGPGVKE EJCPIGU OKIJV QEEWT, YJKEJ OCMGU KV KORQTVCPV VQ RGTHQTO C RJ[UKQNQIKECN EJCTCEVGTKUCVKQP QH VJG TGUWNVKPI UVTCKP þûNUUQP CPF 0KGNUGP 2ýýý). 5WEJ C EJCTCEVGTKUCVKQP OKIJV DG FQPG YKVJ &0# CTTC[U D[ J[DTKFKUKPI NCDGNNGF O40# QT UKORN[ IGPQOKE &0#.
(KI. þ. 6JG UVTCVGI[ QH KPXGTUG OGVCDQNKE GPIKPGGTKPI WUKPI &0# CTTC[U. ôGPGU, YJKEJ ECWUG C FGUKTGF RJGPQV[RG KP G.I. C JKIJ RTQFWEKPI UVTCKP, OC[ DG KFGPVKHKGF VJTQWIJ EQORCTKUQP QH VTCPUETKRVKQP RTQHKNGU HTQO UVTCKPU YKVJ VJG FGUKTGF RJGPQV[RG, UWEJ CU ENCUUKECNN[ KORTQXGF/JKIJ RTQFWEKPI UVTCKPU YKVJ EQTTGURQPFKPI YKNF V[RG UVTCKPU. 6JG KFGPVKHKGF IGPGU ECP VJGP DG OCPKRWNCVGF D[ IGPGVKE GPIKPGGTKPI KP CPQVJGT UVTCKP VQ CEJKGXG C UKOKNCT RJGPQV[RG.
3óý
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
6QFC[, VTCPUETKRVQOG CPCN[UKU KU RTQDCDN[ VJG OQUV WUGHWN HWPEVKQPCN IGPQOKEU VQQN HQT FKTGEV WVKNKUCVKQP KP OGVCDQNKE GPIKPGGTKPI, DWV CPCN[UKU QH VJG QVJGT ÂQOGU• ECP CNUQ DG HQTGUGGP CU WUGHWN VQQNU. ìWCPVKVCVKXG RTQVGQOG CPCN[UKU ECP RQVGPVKCNN[ DG WUGF KPUVGCF QH VTCPUETKRVQOG CPCN[UKU CPF YKNN QHHGT C JKIJGT NGXGN QH KPHQTOCVKQP ENQUGT VQ VJG RJGPQV[RG. *QYGXGT, DGECWUG RTQVGQOG CPCN[UKU UVKNN KP OCP[ YC[U KU KP KVU KPHCPE[ CPF JCU PQV DGEQOG VTWN[ INQDCN [GV, VTCPUETKRVQOG CPCN[UKU KU UVKNN VJG OQUV WUGHWN VQQN. %QORCTCVKXG UGSWGPEG CPCN[UKU QH NCDQTCVQT[ UVTCKPU YKVJ KPFWUVTKCN RTQFWEVKQP UVTCKPU OKIJV DG XCNWCDNG KP WPFGTUVCPFKPI VJG RTQDNGOU VJCV UQOGVKOGU CTG GPEQWPVGTGF YJGP VT[KPI VQ VTCPUHGT OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU HTQO NCDQTCVQT[ UVTCKPU VQ RTQFWEVKQP UVTCKPU. $GVVGT MPQYNGFIG QH VJG IGPQOKE FKHHGTGPEGU OC[ GCUG CPF CNNQY GUVKOCVKQP QH VJG HGCUKDKNKV[ QH VTCPUHGTTKPI OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU. úGVCDQNQOG CPCN[UKU OKIJV CKF OGVCDQNKE GPIKPGGTKPI D[ IKXKPI KPUKIJV KPVQ OGVCDQNKE HWPEVKQP QH OWVCVGF IGPGU KP OWVCPVU D[ EQORCTKUQP YKVJ C TGHGTGPEG UVTCKP. +V OC[ CNUQ RTQXKFG KPFKTGEV GXKFGPEG QH EJCPIGU KP HNWZ TGIWNCVKQP COQPI FKHHGTGPV EQPFKVKQPU KH EJCPIGU KP HNWZ VJTQWIJ C RCVJYC[ CNVGT VJG SWCUK-UVGCF[-UVCVG EQPEGPVTCVKQPU QH CNN QT C UWDUGV QH VJG KPVGTOGFKCVGU KP VJG RCVJYC[ þûNKXGT GV CN. 2ýý2). ôNQDCN OWVCPV EQNNGEVKQPU OC[ DGEQOG C XGT[ RQYGTHWN VQQN KP OGVCDQNKE GPIKPGGTKPI DGECWUG VJG[ CNNQY UGCTEJKPI HQT UVTCKPU YKVJ C FGUKTGF RJGPQV[RG COQPI VJQWUCPF QH OWVCPVU UKOWNVCPGQWUN[ FKTGEVN[ KP VJG GPXKTQPOGPV YJGTG C IGPGVKE EJCPIG KU FGUKTGF. 6JKU VGEJPQNQI[ OKIJV DG WUGF HQT QRVKOKUCVKQP QH JGVGTQNQIQWU RTQVGKP RTQFWEVKQP D[ HKTUV VTCPUHQTOKPI C DCTEQFGF UVTCKP EQNNGEVKQP YKVJ VJG IGPG EQFKPI HQT VJG JGVGTQNQIQWU RTQVGKP CPF VJGP UGCTEJ HQT UVTCKPU YKVJ CP KORTQXGF RJGPQV[RG. 11.5.1 0GGF HQT KPVGITCVGF CRRTQCEJ ûPG QH VJG DKIIGUV EJCNNGPIGU KP HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI KU VQ EQODKPG FCVC HTQO CPCN[UKU QH VJG FKHHGTGPV ÂQOGU•. $KQKPHQTOCVKEU KU CDUQNWVGN[ GUUGPVKCN HQT KPVGITCVKPI FCVC HTQO VJG FKHHGTGPV EGNNWNCT NGXGNU CPF VQ CNNQY CP WPFGTUVCPFKPI QH VJGKT KPVGTCEVKQPU CPF TGIWNCVKQP. *WIG COQWPVU QH FCVC CTG CEJKGXGF HTQO LWUV C UKPING IGPQOG-YKFG CPCN[UKU. +P QTFGT VQ OCMG DKQNQIKECN UGPUG QWV QH VJG FCVC VQ KFGPVKH[ PGY VCTIGVU HQT OGVCDQNKE GPIKPGGTKPI CPF VQ CUUKIP HWPEVKQP VQ QTRJCP IGPGU, DKQKPHQTOCVKEU JCU CNTGCF[ UJQYGF KVU WUGHWNPGUU. *QYGXGT, UQ HCT, DKQKPHQTOCVKEU JCU OQUVN[ DGGP HQEWUUKPI QP FCVC OKPKPI HQT MPQYNGFIG FKUEQXGT[. 6JKU KU PQY EJCPIKPI CU OQTG CVVGPVKQP KU RWV QP UKOWNCVKQP-DCUGF CRRTQCEJGU þ÷KVCPQ 2ýý2). 4GEGPVN[, C IGPQOG-UECNG TGEQPUVTWEVKQP QH VJG S. cerevisiae OGVCDQNKE PGVYQTM YCU RWDNKUJGF þ({TUVGT GV CN. 2ýý3). ùKVJ UWEJ C TGEQPUVTWEVGF PGVYQTM, KV KU RQUUKDNG VQ UGV WR OGVCDQNKE OQFGNU VJCV ECP DG WUGF VQ GXCNWCVG CPF UKOWNCVG VJG RGTHQTOCPEG QH VJG OGVCDQNKE PGVYQTM þ%QXGTV GV CN. 2ýý1). 6JWU, VJGUG FGVCKNGF OGVCDQNKE OQFGNU ECP DG WUGF VQ FGUKIP OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU, DWV VJG[ ECP CNUQ DG WUGF VQ UVWF[ VJG EQPUGSWGPEGU QH FGNGVKPI KPFKXKFWCN IGPGU KP VJG QTICPKUO þ%QXGTV GV CN. 2ýý1). *GTGD[, VJG OQFGNU OC[ DG WUGF VQIGVJGT YKVJ GZRGTKOGPVCN FCVC VQ ICKP PGY KPUKIJV KPVQ VJG QRGTCVKQP QH C EQORNGZ OGVCDQNKE PGVYQTM. *QYGXGT, C FTCYDCEM QH VJG EWTTGPV OQFGNU KU VJCV VJG[ FQ PQV EQPUKFGT
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ó1
TGIWNCVKQP CV VJG NGXGN QH VJG VTCPUETKRVQOG, RTQVGQOG, CPF KPVGTCEVQOG, CPF KV YKNN DG C OCLQT VCUM VQ GZVGPF VJG OQFGNU YKVJ VJGUG HGCVWTGU, DWV UQOG KPKVKCVKXGU CTG QPIQKPI VQ OQFGN EQORNGVG TGIWNCVQT[ UVTWEVWTGU KP S. cerevisiae þ©PF[ CPF $TGPV 2ýý1). #U FKUEWUUGF CDQXG, HWPEVKQPCN IGPQOKEU ECP EQPVTKDWVG VQ OGVCDQNKE GPIKPGGTKPI YKVJ VQQNU HQT CPCN[UKU CPF RJGPQV[RKE EJCTCEVGTKUCVKQP CPF KPETGCUGF MPQYNGFIG CDQWV EGNNWNCT HWPEVKQPU. *QYGXGT, OGVCDQNKE GPIKPGGTKPI CNUQ EQPVTKDWVG VQ HWPEVKQPCN IGPQOKEU YKVJ KVU VTCFKVKQP CPF GZRGTKGPEG KP VCMKPI C U[UVGOU CRRTQCEJ YKVJ HQEWU QP VJG RJGPQV[RG QH VJG EGNN þ(KIWTG 7). #P KORQTVCPV QWVEQOG QH VJG OGVCDQNKE GPIKPGGTKPI E[ENG KU VJG GNWEKFCVKQP QH HWPEVKQP-IGPQV[RG TGNCVKQPUJKRU, CPF KP VJKU UGPUG, OGVCDQNKE GPIKPGGTKPI JCU DGGP CV VJG HQTGHTQPV QH HWPEVKQPCN IGPQOKEU þ5VCHHQTF CPF 5VGRJCPQRQWNQU 2ýý1). úGVCDQNKE GPIKPGGTKPI ECP CNUQ EQPVTKDWVG YKVJ KVU HQEWU QP WUKPI FGHKPGF GZRGTKOGPVCN EQPFKVKQPU, YJKEJ CTG PGEGUUCT[ HQT RTQRGT CPCN[UKU QH RJGPQV[RG. úCP[ INQDCN UVWFKGU KP HWPEVKQPCN IGPQOKEU CTG FQPG QP EWNVWTGU ITQYP KP EQORNGZ OGFKC HQT CP WPURGEKHKGF RGTKQF þ5EJCGEJVGT CPF +PITCJCO 2ýý2). ñPFGT UWEJ EQPFKVKQPU, VJG EQORQUKVKQP QH VJG OGFKWO, CPF VJGTGHQTG, VJG RJ[UKQNQIKECN UVCVG QH VJG EGNNU KU PQV YGNN FGHKPGF, CPF VJKU ECP IKXG RTQDNGOU KP KPVGTRTGVCVKQP QH VJG QDVCKPGF FCVC.
11.þ (WVWTG RTQURGEVU (WPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI CTG VYQ TGUGCTEJ CTGCU YKVJ RCTVN[ QXGTNCRRKPI KPVGTGUVU. 6JG EQPPGEVKQP QH HWPEVKQPCN IGPQOKEU YKVJ OGVCDQNKE GPIKPGGTKPI KU DCUGF QP C DK-FKTGEVKQPCN KPVGTCEVKQP KP YJKEJ VJG HWPEVKQPCN IGPQOKEU VGEJPQNQIKGU YKNN JGNR FGXGNQR VQQNU CPF TGCIGPVU þIGPGU, RTQOQVGTU GVE.) PGGFGF HQT RCVJYC[ OCPKRWNCVKQP, DWV CNUQ DGECWUG OGVCDQNKE GPIKPGGTKPI RTQXKFGU C EQPXGPKGPV HTCOGYQTM HQT VJG CPCN[UKU CPF KPVGITCVKQP QH UWEJ FCVC þ0KGNUGP 2ýý1ø 5VCHHQTF CPF 5VGRJCPQRQWNQU 2ýý1). 6JG WUG QH &0# CTTC[U CPF VTCPUETKRVQOG
(KI. 7. 6JG KPVGTCEVKQP DGVYGGP HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI KU DKFKTGEVKQPCN. (WPEVKQPCN IGPQOKEU ECP EQPVTKDWVG VQ OGVCDQNKE GPIKPGGTKPI YKVJ VQQNU HQT YJQNG-EGNN CPCN[UKU. úGVCDQNKE GPIKPGGTKPI ECP EQPVTKDWVG VQ HWPEVKQPCN IGPQOKEU YKVJ KVU HQEWU QP RJGPQV[RG þVJG GPVKTG U[UVGO) CPF YKVJ KVU HQEWU QP WUKPI FGHKPGF GZRGTKOGPVCN EQPFKVKQPU.
3ó2
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
CPCN[UKU KU CNTGCF[ HKPFKPI KVU RNCEG CU C VQQN KP OGVCDQNKE GPIKPGGTKPI CU KNNWUVTCVGF D[ VJG GZCORNGU KP VJKU EJCRVGT, CPF KP VJG HWVWTG, YG GZRGEV VJCV OCP[ UVWFKGU YKNN WUG EQODKPCVKQPU QH VJG FKHHGTGPV CPCN[UKU OGVJQFU HQT VJG ÂQOGU•. 6JGTG YKNN DG JWIG FGXGNQROGPVU CPF KORTQXGOGPVU QH CPCN[VKECN OGVJQFU KP RCTVKEWNCT KP RTQVGQOG CPCN[UKU, CPF VQIGVJGT YKVJ KORTQXGF RGTHQTOCPEG QH VJG VGEJPKSWGU QH HWPEVKQPCN IGPQOKEU, VJG[ YKNN RTQFWEG OQTG CEEWTCVG FCVC. 6JKU YKNN GPCDNG KORTQXGF TGUQNWVKQP QH VJG OCP[ FKHHGTGPV RCVJYC[U QRGTCVKPI KP NKXKPI EGNNU CPF VJKU OGEJCPKUVKE KPHQTOCVKQP OC[ DG WUGF VQ TGEQPUVTWEV EQORNGVG TGIWNCVQT[ UVTWEVWTGU. 6JKU KPHQTOCVKQP YKNN RNC[ CP KORQTVCPV TQNG KP FGUKIPKPI PGY OGVCDQNKE GPIKPGGTKPI UVTCVGIKGU KP VJG HWVWTG.
#EMPQYNGFIGOGPVU ûWT URGEKCN VJCPMU IQ VQ ÷CURGT úòNNGT HQT ECTGHWNN[ TGCFKPI CPF EQOOGPVKPI VJKU OCPWUETKRV. 6JG TGUGCTEJ YQTM QP HWPEVKQPCN IGPQOKEU CV %GPVGT HQT öTQEGUU $KQVGEJPQNQI[ KU HQWPFGF XKC VJG &CPKUJ $KQVGEJPQNQI[ +PUVTWOGPV %GPVGT þ&#$+%).
ýGHGTGPEGU #GDGTUQNF 4, ôQQFNGVV &4 þ2ýý1) úCUU URGEVTQOGVT[ KP RTQVGQOKEU. %JGO 4GX 1ý1:2õ929ó #TKUVKFQW #, öGPVVKNk ú þ2ýýý) úGVCDQNKE GPIKPGGTKPI CRRNKECVKQPU VQ TGPGYCDNG TGUQWTEG WVKNK\CVKQP. %WTT ûRKP $KQVGEJPQN 11:187-198 $CEMJWU ï©, &G4KUK ÿ, $TQYP öû, $KUUQP ï( þ2ýý1) (WPEVKQPCN IGPQOKE CPCN[UKU QH C EQOOGTEKCN YKPG UVTCKP QH Saccharomyces cerevisiae WPFGT FKHHGTKPI PKVTQIGP EQPFKVKQPU. (©ú5 ;GCUV 4GUGCTEJ 1:111-12ó $CKNG[ ÿ© þ1991) 6QYCTF C UEKGPEG QH OGVCDQNKE GPIKPGGTKPI. 5EKGPEG 2ó2:1õõ8-1õ7ó $CKNG[ ÿ© þ1999) ïGUUQPU HTQO OGVCDQNKE GPIKPGGTKPI HQT HWPEVKQPCN IGPQOKEU CPF FTWI FKUEQXGT[. 0CV $KQVGEJPQN 17:õ1õ-õ18 $CKNG[ ÿ©, 5JWTNCVK #, *CV\KOCPKMCVKU í, ïGG ÷, 4GPPGT ù#, 6UCK ö5 þ199õ) +PXGTUG OGVCDQNKE GPIKPGGTKPI: # UVTCVGI[ HQT FKTGEVGF IGPGVKE GPIKPGGTKPI QH WUGHWN RJGPQV[RGU. $KQVGEJPQN $KQGPI ó2:1ý9-121 $GPPGVV %$, ïGYKU ï÷, ÷CTVJKMG[CP ô, ïQDCEJGX ÷5, ÿKP ;*, 5VGTNKPI ÿ(, 5PKRG ÿ4, 4GUPKEM ú# þ2ýý1) ôGPGU TGSWKTGF HQT KQPK\KPI TCFKCVKQP TGUKUVCPEG KP [GCUV. 0CV ôGPGV 29:ü2õ-ü3ü $KTTGNN ôù, ôKCGXGT ô, %JW #ú, &CXKU 4ù, $TQYP ÿú þ2ýý1) # IGPQOG-YKFG UETGGP KP Saccharomyces cerevisiae HQT IGPGU CHHGEVKPI ñí TCFKCVKQP UGPUKVKXKV[. öTQE 0CVN #ECF 5EK ñ5# 98:12õý8-12õ13 $KTTGNN ôù, $TQYP ÿ#, ùW *+, ôKCGXGT ô, %JW #ú, &CXKU 4ù, $TQYP ÿú þ2ýý2) 6TCPUETKRVKQPCN TGURQPUG QH Saccharomyces cerevisiae VQ &0#-FCOCIKPI CIGPVU FQGU PQV KFGPVKH[ VJG IGPGU VJCV RTQVGEV CICKPUV VJGUG CIGPVU. öTQE 0CVN #ECF 5EK ñ5# 99:87788783 $NCEMUVQEM ù, úCPP ú þ2ýýý) öTQVGQOKEU: # 6TGPFU ôWKFG. ©NUGXKGT, #OUVGTFCO $TGPV 4 þ2ýýý) ôGPQOKE DKQNQI[. %GNN 1ýý:1õ9-183
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ó3
$WTPU 0, ôTKOYCFG $, 4QUU-úCEFQPCNF ö$, %JQK ©;, (KPDGTI ÷, 4QGFGT ô5, 5P[FGT ú þ199ü) ïCTIG-UECNG CPCN[UKU QH IGPG GZRTGUUKQP, RTQVGKP NQECNK\CVKQP, CPF IGPG FKUTWRVKQP KP Saccharomyces cerevisiae. ôGPGU &GX 8:1ý87-11ýó $WUUGOCMGT *ÿ, ïK *, 5KIIKC ©& þ2ýý1) 4GIWNCVQT[ GNGOGPV FGVGEVKQP WUKPI EQTTGNCVKQP YKVJ GZRTGUUKQP. 0CV ôGPGV 27:1õ7-171 %CXCNKGTK &, 6QYPUGPF ÿö, *CTVN &ï þ2ýýý) úCPKHQNF CPQOCNKGU KP IGPG GZRTGUUKQP KP C XKPG[CTF KUQNCVG QH Saccharomyces cerevisiae TGXGCNGF D[ &0# OKETQCTTC[ CPCN[UKU. öTQE 0CVN #ECF 5EK ñ5# 97:123õ9-1237ü %JQ 4ÿ, %CORDGNN úÿ, ùKP\GNGT ©#, 5VGKPOGV\ ï, %QPYC[ #, ùQFKEMC ï, ùQNHUDGTI 6ô, ôCDTKGNKCP #©, ïCPFUOCP &, ïQEMJCTV &ÿ, &CXKU 4ù þ1998) # IGPQOG-YKFG VTCPUETKRVKQPCN CPCN[UKU QH VJG OKVQVKE EGNN E[ENG. úQN %GNN 2:õó-73 %JQVCPK ô, &QFIG 6, *UW #, ÷WOCT ú, ïC&WEC 4, 6TKODWT &, ùG[NGT ù, 5CPHQTF ÷ þ2ýýý) 6JG EQOOGTEKCN RTQFWEVKQP QH EJGOKECNU WUKPI RCVJYC[ GPIKPGGTKPI. $KQEJKO $KQRJ[U #EVC 1óü3:ü3ü-üóó %JTKUVGPUGP $, 0KGNUGP ÿ þ2ýýý) úGVCDQNKE PGVYQTM CPCN[UKU. # RQYGTHWN VQQN KP OGVCDQNKE GPIKPGGTKPI. #FX $KQEJGO ©PI $KQVGEJPQN õõ:2ý9-231 %JW 5, &G4KUK ÿ, ©KUGP ú, úWNJQNNCPF ÿ, $QVUVGKP &, $TQYP öû, *GTUMQYKV\ + þ1998) 6JG VTCPUETKRVKQPCN RTQITCO QH URQTWNCVKQP KP DWFFKPI [GCUV. 5EKGPEG 282:õ99-7ýó %JWC ô, 6CTKECPK ï, 5VCPING ù, ;QWPI öô þ2ýýý) +PUGTVKQPCN OWVCIGPGUKU DCUGF QP KNNGIKVKOCVG TGEQODKPCVKQP KP Schizosaccharomyces pombe. 0WENGKE #EKFU 4GU 28:©ó3 %NKHVGP ö(, *KNNKGT ïù, (WNVQP ï, ôTCXGU 6, úKPGT 6, ôKUJ ù4, ùCVGTUVQP 4*, ÿQJPUVQP ú þ2ýý1) 5WTXG[KPI Saccharomyces IGPQOGU VQ KFGPVKH[ HWPEVKQPCN GNGOGPVU D[ EQORCTCVKXG &0# UGSWGPEG CPCN[UKU. ôGPQOG 4GU 11:117ó-118õ %QXGTV úù, 5EJKNNKPI %*, (COKNK +, ©FYCTFU ÿ5, ôQT[CPKP ++, 5GNMQX ©, öCNUUQP $û þ2ýý1) úGVCDQNKE OQFGNKPI QH OKETQDKCN UVTCKPU KP UKNKEQ. 6TGPFU $KQEJGO 5EK 2õ:17918õ &GNPGTK &, $TCPEKC (ï, ûNKXGT 5ô þ2ýý1) 6QYCTFU C VTWN[ KPVGITCVKXG DKQNQI[ VJTQWIJ VJG HWPEVKQPCN IGPQOKEU QH [GCUV. %WTT ûRKP $KQVGEJPQN 12:87-91 &G4KUK ÿï, +[GT í4, $TQYP öû þ1997) ©ZRNQTKPI VJG OGVCDQNKE CPF IGPGVKE EQPVTQN QH IGPG GZRTGUUKQP QP C IGPQOKE UECNG. 5EKGPEG 278:õ8ý-õ8õ >OKV\CMKU ©6, 4G[OQPF #, ï[NG 4, 5ECOWHHC 0, ñENC %, &GWVUEJ 5, 5VGXGPUQP $ÿ, (NGIGN í, $WEJGT ö, ÿQPIGPGGN %í, #PVQPCTCMKU 5© þ2ýý2) 0WOGTQWU RQVGPVKCNN[ HWPEVKQPCN DWV PQP-IGPKE EQPUGTXGF UGSWGPEGU QP JWOCP EJTQOQUQOG 21. 0CVWTG ü2ý:ó78ó82 &QTPGT #ÿ, ÷CWHOCP 4ÿ þ199ü) 6JG NGXGNU QH GPFQRNCUOKE TGVKEWNWO RTQVGKPU CPF #6ö CHHGEV HQNFKPI CPF UGETGVKQP QH UGNGEVKXG RTQVGKPU. $KQNQIKECNU 22:1ý3-112 ©INKPVQP ÿú, *GKPTKEJ #ÿ, öQNNPKV\ #ö, ïCPITKFIG ö, *GPUEJMG ö#, FG $CTTQU ïú þ2ýý2) &GETGCUKPI CEGVKE CEKF CEEWOWNCVKQP D[ C IN[EGTQN QXGTRTQFWEKPI UVTCKP QH Saccharomyces cerevisiae D[ FGNGVKPI VJG ALD6 CNFGJ[FG FGJ[FTQIGPCUG IGPG. ;GCUV 19:29ó3ý1 ©KUGP ú$, 5RGNNOCP ö6, $TQYP öû, $QVUVGKP & þ1998) %NWUVGT CPCN[UKU CPF FKURNC[ QH IGPQOG-YKFG GZRTGUUKQP RCVVGTPU. öTQE 0CVN #ECF 5EK ñ5# 9ó:1ü8õ3-1ü8õ8 ©PF[ &, $TGPV 4 þ2ýý1) úQFGNNKPI EGNNWNCT DGJCXKQWT. 0CVWTG üý9 5WRRN:391-39ó (KGJP û þ2ýý1) %QODKPKPI IGPQOKEU, OGVCDQNQOG CPCN[UKU, CPF DKQEJGOKECN OQFGNNKPI VQ WPFGTUVCPF OGVCDQNKE PGVYQTMU. %QOR (WPEV ôGPQO 2:1óó-1õ8 (KGJP û þ2ýý2) úGVCDQNQOKEU--VJG NKPM DGVYGGP IGPQV[RGU CPF RJGPQV[RGU. öNCPV úQN $KQN ü8:1óó-171
3óü
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
(KGNFU 5, 5QPI û þ1989) # PQXGN IGPGVKE U[UVGO VQ FGVGEV RTQVGKP-RTQVGKP KPVGTCEVKQPU. 0CVWTG 3üý:2üó-2üõ ({TUVGT ÿ, ôQODGTV #÷, 0KGNUGP ÿ þ2ýý2) # HWPEVKQPCN IGPQOKEU CRRTQCEJ WUKPI OGVCDQNQOKEU CPF KP UKNKEQ RCVJYC[ CPCN[UKU. $KQVGEJPQN $KQGPI 79:7ý3-712 ({TUVGT ÿ, (COKNK +, (W ö, öCNUUQP $·, 0KGNUGP ÿ þ2ýý3) ôGPQOG-UECNG TGEQPUVTWEVKQP QH VJG Saccharomyces cerevisiae OGVCDQNKE PGVYQTM. ôGPQOG 4GU 13:2üü-2ó3 (TKGFOCP 0, ïKPKCN ú, 0CEJOCP +, öG'GT & þ2ýýý) ñUKPI $C[GUKCP PGVYQTMU VQ CPCN[\G GZRTGUUKQP FCVC. ÿ %QORWV $KQN 7:õý1-õ2ý (WVEJGT $, ïCVVGT ô+, úQPCTFQ ö, úEïCWIJNKP %5, ôCTTGNU ÿ+ þ1999) # UCORNKPI QH VJG [GCUV RTQVGQOG. úQN %GNN $KQN 19:73ó7-73õ8 ôCUEJ #ö, 5RGNNOCP ö6, ÷CQ %ú, %CTOGN-*CTGN û, ©KUGP ú$, 5VQT\ ô, $QVUVGKP &, $TQYP öû þ2ýýý) ôGPQOKE GZRTGUUKQP RTQITCOU KP VJG TGURQPUG QH [GCUV EGNNU VQ GPXKTQPOGPVCN EJCPIGU. úQN $KQN %GNN 11:ü2ü1-ü2ó7 ôCUEJ #ö, *WCPI ú, úGV\PGT 5, $QVUVGKP &, ©NNGFIG 5ÿ, $TQYP öû þ2ýý1) ôGPQOKE GZRTGUUKQP TGURQPUGU VQ &0#-FCOCIKPI CIGPVU CPF VJG TGIWNCVQT[ TQNG QH VJG [GCUV #64 JQOQNQI úGE1R. úQN $KQN %GNN 12:2987-3ýý3 ôCXKP #%, $QUEJG ú, ÷TCWUG 4, ôTCPFK ö, úCT\KQEJ ú, $CWGT #, 5EJWNV\ ÿ, 4KEM ÿú, úKEJQP #ú, %TWEKCV %ú, 4GOQT ú, *QHGTV %, 5EJGNFGT ú, $TCLGPQXKE ú, 4WHHPGT *, úGTKPQ #, ÷NGKP ÷, *WFCM ú, &KEMUQP &, 4WFK 6, ôPCW í, $CWEJ #, $CUVWEM 5, *WJUG $, ïGWVYGKP %, *GWTVKGT ú#, %QRNG[ 44, ©FGNOCPP #, ìWGTHWTVJ ©, 4[DKP í, &TGYGU ô, 4CKFC ú, $QWYOGGUVGT 6, $QTM ö, 5GTCRJKP $, ÷WUVGT $, 0GWDCWGT ô, 5WRGTVK-(WTIC ô þ2ýý2) (WPEVKQPCN QTICPK\CVKQP QH VJG [GCUV RTQVGQOG D[ U[UVGOCVKE CPCN[UKU QH RTQVGKP EQORNGZGU. 0CVWTG ü1ó:1ü1-1ü7 ôKCGXGT ô GV CN. þ2ýý2) (WPEVKQPCN RTQHKNKPI QH VJG Saccharomyces cerevisiae IGPQOG. 0CVWTG ü18:387-391 ôKNN 46, ùKNFV 5, ;CPI ;6,
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3óó
*CNNDQTP ÿ, ùCNHTKFUUQP ú, #KTCMUKPGP ñ, ûLCOQ *, *CJP-*kIGTFCN $, öGPVVKNC ú, ÷GTCUPGP 5 þ1991) ð[NKVQN RTQFWEVKQP D[ TGEQODKPCPV Saccharomyces cerevisiae. $KQVGEJPQNQI[ þ0;) 9:1ý9ý-1ý9ó *COOQPF %, *GNGPKWU # þ199ó) ìWCNKV[ EQPVTQN KP VJG UGETGVQT[ RCVJYC[. %WTT ûRKP %GNN $KQN 7:ó23-ó29 *CWUGT 0%, (GNNGPDGTI ÷, ôKN 4, $CUVWEM 5, *QJGKUGN ÿ&, öoTG\-ûTVKP ÿ© þ2ýý1) ùJQNG IPQOG CPCN[UKU QH C YKPG [GCUV UVTCKP. %QOR (WPEV ôGPQO 2:õ9-79 *GNNGT úÿ þ2ýý2) &0# OKETQCTTC[ VGEJPQNQI[: FGXKEGU, U[UVGOU, CPF CRRNKECVKQPU. #PPW 4GX $KQOGF ©PI ü:129-1ó3 *Q 0ù, %JGP <, $TCKPCTF #ö þ1998) ôGPGVKECNN[ GPIKPGGTGF Saccharomyces [GCUV ECRCDNG QH GHHGEVKXG EQHGTOGPVCVKQP QH INWEQUG CPF Z[NQUG. #RRN ©PXKTQP úKETQDKQN õü:18ó218ó9 *Q ;, ôTWJNGT #, *GKNDWV #, $CFGT ô&, úQQTG ï, #FCOU 5ï, úKNNCT #, 6C[NQT ö, $GPPGVV ÷, $QWVKNKGT ÷, ;CPI ï, ùQNVKPI %, &QPCNFUQP +, 5EJCPFQTHH 5, 5JGYPCTCPG ÿ, íQ ú, 6CIICTV ÿ, ôQWFTGCWNV ú, úWUMCV $, #NHCTCPQ %, &GYCT &, ïKP <, úKEJCNKEMQXC ÷, ùKNNGOU #4, 5CUUK *, 0KGNUGP ö#, 4CUOWUUGP ÷ÿ, #PFGTUGP ÿ4, ÿQJCPUGP ï©, *CPUGP ï*, ÿGURGTUGP *, öQFVGNGLPKMQX #, 0KGNUGP ©, %TCYHQTF ÿ, öQWNUGP í, 5QTGPUGP $&, úCVVJKGUGP ÿ, *GPFTKEMUQP 4%, ôNGGUQP (, öCYUQP 6, úQTCP ú(, &WTQEJGT &, úCPP ú, *QIWG %ù, (KIG[U &, 6[GTU ú þ2ýý2) 5[UVGOCVKE KFGPVKHKECVKQP QH RTQVGKP EQORNGZGU KP Saccharomyces cerevisiae D[ OCUU URGEVTQOGVT[. 0CVWTG ü1ó:18ý-183 *QHHOCP %5, ùGNVQP 4 þ2ýýý) úWVCIGPGUKU CPF IGPG ENQPKPI KP Schizosaccharomyces pombe WUKPI PQPJQOQNQIQWU RNCUOKF KPVGITCVKQP CPF TGUEWG. $KQVGEJPKSWGU 28:ó32-õ, ó38, óüý *QTCM %©, 5P[FGT ú þ2ýý2) ôNQDCN CPCN[UKU QH IGPG GZRTGUUKQP KP [GCUV. (WPEV +PVGIT ôGPQOKEU 2:171-18ý *WIJGU ÿ&, ©UVGR öù, 6CXC\QKG 5, %JWTEJ ôú þ2ýýý) %QORWVCVKQPCN KFGPVKHKECVKQP QH EKUTGIWNCVQT[ GNGOGPVU CUUQEKCVGF YKVJ ITQWRU QH HWPEVKQPCNN[ TGNCVGF IGPGU KP Saccharomyces cerevisiae. ÿ úQN $KQN 29õ:12ýó-121ü +FGMGT 6, 6JQTUUQP í, 4CPKUJ ÿ#, %JTKUVOCU 4, $WJNGT ÿ, ©PI ÿ÷, $WOICTPGT 4, ôQQFNGVV &4, #GDGTUQNF 4, *QQF ï þ2ýý1) +PVGITCVGF IGPQOKE CPF RTQVGQOKE CPCN[UGU QH C U[UVGOCVKECNN[ RGTVWTDGF OGVCDQNKE PGVYQTM. 5EKGPEG 292:929-93ü +VQ 6, 6CUJKTQ ÷, úWVC 5, û\CYC 4, %JKDC 6, 0KUJK\CYC ú, ;COCOQVQ ÷, ÷WJCTC 5, 5CMCMK ; þ2ýýý) 6QYCTF C RTQVGKP-RTQVGKP KPVGTCEVKQP OCR QH VJG DWFFKPI [GCUV: # EQORTGJGPUKXG U[UVGO VQ GZCOKPG VYQ-J[DTKF KPVGTCEVKQPU KP CNN RQUUKDNG EQODKPCVKQPU DGVYGGP VJG [GCUV RTQVGKPU. öTQE 0CVN #ECF 5EK ñ5# 97:11ü3-11ü7 +[GT í4, *QTCM %©, 5ECHG %5, $QVUVGKP &, 5P[FGT ú, $TQYP öû þ2ýý1) ôGPQOKE DKPFKPI UKVGU QH VJG [GCUV EGNN-E[ENG VTCPUETKRVKQP HCEVQTU 5$( CPF ú$(. 0CVWTG üý9:ó33-ó38 ÿGPUGP ïÿ, ÷PWFUGP 5 þ2ýýý) #WVQOCVKE FKUEQXGT[ QH TGIWNCVQT[ RCVVGTPU KP RTQOQVGT TGIKQPU DCUGF QP YJQNG EGNN GZRTGUUKQP FCVC CPF HWPEVKQPCN CPPQVCVKQP. $KQKPHQTOCVKEU 1õ:32õ-333 ÿGRRUUQP ú, ÿQJCPUUQP $, *CJP-*kIGTFCN $, ôQTYC-ôTCWUNWPF ú( þ2ýý2) 4GFWEGF QZKFCVKXG RGPVQUG RJQURJCVG RCVJYC[ HNWZ KP TGEQODKPCPV Z[NQUG- WVKNK\KPI Saccharomyces cerevisiae UVTCKPU KORTQXGU VJG GVJCPQN [KGNF HTQO Z[NQUG. #RRN ©PXKTQP úKETQDKQN õ8:1õýü-1õý9 ÿQJCPUUQP $, %JTKUVGPUUQP %, *QDNG[ 6, *CJP-*kIGTFCN $ þ2ýý1) ð[NWNQMKPCUG QXGTGZRTGUUKQP KP VYQ UVTCKPU QH Saccharomyces cerevisiae CNUQ GZRTGUUKPI Z[NQUG TGFWEVCUG CPF Z[NKVQN FGJ[FTQIGPCUG CPF KVU GHHGEV QP HGTOGPVCVKQP QH Z[NQUG CPF NKIPQEGNNWNQUKE J[FTQN[UCVG. #RRN ©PXKTQP úKETQDKQN õ7:ü2ü9-ü2óó
3óõ
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
÷GCNG[ ÿ6, ïKW ï, 5CPVK &í, $GVNCEJ ú%, $CTT öÿ þ1998) öTQFWEVKQP QH C RQN[MGVKFG PCVWTCN RTQFWEV KP PQPRQN[MGVKFG-RTQFWEKPI RTQMCT[QVKE CPF GWMCT[QVKE JQUVU. öTQE 0CVN #ECF 5EK ñ5# 9ó:óýó-óý9 ÷GNGU 5, XCP FGT ïú, ©KUGP ú$ þ2ýý2) +FGPVKHKECVKQP QH TGIWNCVQT[ GNGOGPVU WUKPI C HGCVWTG UGNGEVKQP OGVJQF. $KQKPHQTOCVKEU 18:11õ7-117ó ÷KVCPQ * þ2ýý2) %QORWVCVKQPCN U[UVGOU DKQNQI[. 0CVWTG ü2ý:2ýõ-21ý ÷LGNFUGP 6, öGVVGTUUQP #(, *CEJ ú, &KGTU +, *CXGNWPF 5, *CPUGP ö*, #PFGTUGP #5 þ1997) 5[PVJGVKE NGCFGTU YKVJ RQVGPVKCN $Kö DKPFKPI OGFKCVG JKIJ-[KGNF UGETGVKQP QH EQTTGEVN[ HQNFGF KPUWNKP RTGEWTUQTU HTQO Saccharomyces cerevisiae. öTQVGKP ©ZRT öWTKH 9:33133õ ÷NGKP %ÿ, ûNUUQP ï, 0KGNUGP ÿ þ1998) ôNWEQUG EQPVTQN KP Saccharomyces cerevisiae: VJG TQNG QH úKI1 KP OGVCDQNKE HWPEVKQPU. úKETQDKQNQI[ 1üü þ öV 1):13-2ü ÷PWFUGP 5 þ2ýý2) # DKQNQIKUV IWKFG VQ CPCN[UKU QH &0# OKETQCTTC[ FCVC. ÿQJP ùKNG[ ê 5QPU, 0GY ;QTM ÷QPFQ ÷, úKWTC ;, 5QPG *, ÷QDC[CUJK ÷, +KLKOC * þ1997) *KIJ-NGXGN GZRTGUUKQP QH C UYGGV RTQVGKP, OQPGNNKP, KP VJG HQQF [GCUV Candida utilis. 0CV $KQVGEJPQN 1ó:üó3-üó7 ÷{VVGT ö, #OQTG 4, *QNNGPDGTI %ö, %KTKCE[ ú þ199ý) +UQNCVKQP CPF EJCTCEVGTK\CVKQP QH VJG Pichia stipitis Z[NKVQN FGJ[FTQIGPCUG IGPG, XYL2, CPF EQPUVTWEVKQP QH C Z[NQUG-WVKNK\KPI Saccharomyces cerevisiae VTCPUHQTOCPV. %WTT ôGPGV 18:ü93-óýý ÷{VVGT ö, %KTKCE[ ú þ1993) ð[NQUG (GTOGPVCVKQP D[ Saccharomyces cerevisiae. #RRN úKETQDKQN $KQVGEJPQN 38:77õ-783 ÷WOCT #, 5P[FGT ú þ2ýý1) ©OGTIKPI VGEJPQNQIKGU KP [GCUV IGPQOKEU. 0CV 4GX ôGPGV 2:3ý2-312 ÷WOCT #, #ICTYCN 5, *G[OCP ÿ#, úCVUQP 5, *GKFVOCP ú, öKEEKTKNNQ 5, ñOCPUM[ ï, &TCYKF #, ÿCPUGP 4, ïKW ;, %JGWPI ÷*, úKNNGT ö, ôGTUVGKP ú, 4QGFGT ô5, 5P[FGT ú þ2ýý2) 5WDEGNNWNCT NQECNK\CVKQP QH VJG [GCUV RTQVGQOG. ôGPGU &GX 1õ:7ý7-719 ïGG 6+, 4KPCNFK 0ÿ, 4QDGTV (, ûFQO &6, $CT-ÿQUGRJ <, ôGTDGT ô÷, *CPPGVV 0ú, *CTDKUQP %6, 6JQORUQP %ú, 5KOQP +,
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ó7
0KGNUGP ÿ, ûNUUQP ï þ2ýý2) #P GZRCPFGF TQNG HQT OKETQDKCN RJ[UKQNQI[ KP OGVCDQNKE GPIKPGGTKPI CPF HWPEVKQPCN IGPQOKEU: OQXKPI VQYCTFU U[UVGOU DKQNQI[. (©ú5 ;GCUV 4GU 2:17ó-181 0KUUGP 6ï, ÷KGNNCPF-$TCPFV ú%, 0KGNUGP ÿ, íKNNCFUGP ÿ þ2ýýý) ûRVKOK\CVKQP QH GVJCPQN RTQFWEVKQP KP Saccharomyces cerevisiae D[ OGVCDQNKE GPIKPGGTKPI QH VJG COOQPKWO CUUKOKNCVKQP. úGVCD ©PI 2:õ9-77 ûNKXGT &ÿ, 0KMQNCW $, ùWTVGNG ©5 þ2ýý2) (WPEVKQPCN IGPQOKEU: JKIJ-VJTQWIJRWV O40#, RTQVGKP, CPF OGVCDQNKVG CPCN[UGU. úGVCD ©PI ü:98-1ýõ ûNKXGT 5ô þ1998) +PVTQFWEVKQP VQ HWPEVQPCN CPCN[UKU QH VJG [GCUV IGPQOG. +P: $TQYP #ÿö, 6WKVG ú þGFU) úGVJQFU KP úKETQDKQNQI[: ;GCUV IGPG CPCN[UKU. #ECFGOKE öTGUU, 5CP &KGIQ, RR 1-13• ûNUUQP ï, 0KGNUGP ÿ þ2ýýý) 6JG TQNG QH OGVCDQNKE GPIKPGGTKPI KP VJG KORTQXGOGPV QH Saccharomyces cerevisiae: WVKNK\CVKQP QH KPFWUVTKCN OGFKC. ©P\[OG úKETQD 6GEJPQN 2õ:78ó-792 ûQK 5ï, 5JQGOCMGT &&, $QGMG ÿ& þ2ýý1) # &0# OKETQCTTC[-DCUGF IGPGVKE UETGGP HQT PQPJQOQNQIQWU GPF-LQKPKPI OWVCPVU KP Saccharomyces cerevisiae. 5EKGPEG 29ü:2óó22óóõ ûUVGTICCTF 5 þ2ýýý) úGVCDQNKE GPIKPGGTKPI QH VJG ICNCEVQUG OGVCDQNKUO KP Saccharomyces cerevisiae. 6GEJPKECN ñPKXGTUKV[ QH &GPOCTM ûUVGTICCTF 5, ûNUUQP ï, ÿQJPUVQP ú, 0KGNUGP ÿ þ2ýýýC) +PETGCUKPI ICNCEVQUG EQPUWORVKQP D[ Saccharomyces cerevisiae VJTQWIJ OGVCDQNKE GPIKPGGTKPI QH VJG GAL IGPG TGIWNCVQT[ PGVYQTM. 0CV $KQVGEJPQN 18:1283-128õ ûUVGTICCTF 5, ûNUUQP ï, 0KGNUGP ÿ þ2ýýýD) úGVCDQNKE GPIKPGGTKPI QH Saccharomyces cerevisiae. úKETQDKQN úQN $KQN 4GX õü:3ü-óý ûUVGTICCTF 5, 4QEC %, 4òPPQY $, 0KGNUGP ÿ, ûNUUQP ï þ2ýýýE) öJ[UKQNQIKECN UVWFKGU KP CGTQDKE DCVEJ EWNVKXCVKQPU QH Saccharomyces cerevisiae UVTCKPU JCTDQTKPI VJG MEL1 IGPG. $KQVGEJPQN $KQGPI õ8:2ó2-2ó9 öCPFG[ #, úCPP ú þ2ýýý) öTQVGQOKEU VQ UVWF[ IGPGU CPF IGPQOGU. 0CVWTG üýó:837-8üõ öCP\PGT 5, &TGKGT ï, *CTVOCPP ©, ÷QUVMC 5, 4CRQRQTV 6# þ199ó) öQUVVTCPUNCVKQPCN RTQVGKP VTCPURQTV KP [GCUV TGEQPUVKVWVGF YKVJ C RWTKHKGF EQORNGZ QH 5GE RTQVGKPU CPF ÷CT2R. %GNN 81:óõ1-ó7ý öCRQWVUCMKU ©6 þ1998) ©ZRTGUU VQIGVJGT CPF EQPSWGT. 0CV $KQVGEJPQN 1õ:ü1õ-ü17 öGTTQV ú, 5CINKQEEQ (, úKPK 6, úQPTKDQV %, 5EJPGKFGT ñ, 5JGXEJGPMQ #, úCPP ú, ÿGPQ ö, $QWEJGTKG * þ1999) 6YQ-FKOGPUKQPCN IGN RTQVGKP FCVCDCUG QH Saccharomyces cerevisiae þWRFCVG 1999). ©NGEVTQRJQTGUKU 2ý:228ý-2298 öoTG\-ûTVsP ÿ©, ôCTEKC-úCTVKPG\ ÿ, #NDGTQNC 6ú þ2ýý2) &0# EJKRU HQT [GCUV DKQVGEJPQNQI[. 6JG ECUG QH YKPG [GCUVU. ÿ $KQVGEJPQN 98:227-2ü1 öTKOKI ú, ùKNNKCOU 4ú, ùKP\GNGT ©#, 6GX\CF\G ôô, %QPYC[ #4, *YCPI 5;, &CXKU 4ù, ©URQUKVQ 4© þ2ýýý) 6JG EQTG OGKQVKE VTCPUETKRVQOG KP DWFFKPI [GCUVU. 0CV ôGPGV 2õ:ü1ó-ü23 ìWG ìì, ùKP\GNGT ©# þ2ýý2) ïCTIG-UECNG OWVCIGPGUKU CPF HWPEVKQPCN IGPQOKEU KP [GCUV. (WPEV +PVGIT ôGPQOKEU 2:193-198 4CCOUFQPM ïú, 6GWUKPM $, $TQCFJWTUV &, <JCPI 0, *C[GU #, ùCNUJ ú%, $GTFGP ÿ#, $TKPFNG ÷ú, ÷GNN &$, 4QYNCPF ÿÿ, ùGUVGTJQHH *í, XCP &CO ÷, ûNKXGT 5ô þ2ýý1) # HWPEVKQPCN IGPQOKEU UVTCVGI[ VJCV WUGU OGVCDQNQOG FCVC VQ TGXGCN VJG RJGPQV[RG QH UKNGPV OWVCVKQPU. 0CV $KQVGEJPQN 19:üó-óý
3ó8
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
4GP $, 4QDGTV (, ù[TKEM ÿÿ, #RCTKEKQ û, ÿGPPKPIU ©ô, 5KOQP +,
11 ;GCUV HWPEVKQPCN IGPQOKEU CPF OGVCDQNKE GPIKPGGTKPI: RCUV, RTGUGPV CPF HWVWTG
3ó9
$, úCNRGTVW[ #, 0GWXGINKUG %, û\KGT-÷CNQIGTQRQWNQU û, öQVKGT 5, 5CWTKP ù, 6GMCKC (, 6QHHCPQ-0KQEJG %, ùGUQNQYUMK-ïQWXGN ú, ùKPEMGT ö, ùGKUUGPDCEJ ÿ þ2ýýý) ôGPQOKE GZRNQTCVKQP QH VJG JGOKCUEQO[EGVQWU [GCUVU: 1. # UGV QH [GCUV URGEKGU HQT OQNGEWNCT GXQNWVKQP UVWFKGU. (©$5 ïGVV ü87:3-12 5VCHHQTF &©, 5VGRJCPQRQWNQU ô þ2ýý1) úGVCDQNKE GPIKPGGTKPI CU CP KPVGITCVKPI RNCVHQTO HQT UVTCKP FGXGNQROGPV. %WTT ûRKP úKETQDKQN ü:33õ-3üý 5VGRJCPQRQWNQU ô, #TKUVQFQW #, 0KGNUGP ÿ þ1998) úGVCDQNKE ©PIKPGGTKPI. #ECFGOKE öTGUU +PE, 5CP &KGIQ 5\[RGTUMK 6 þ1998) 13%-0ú4, ú5 CPF OGVCDQNKE HNWZ DCNCPEKPI KP DKQVGEJPQNQI[ TGUGCTEJ. ì 4GX $KQRJ[U 31:ü1-1ýõ 6CMCJCUJK 6, 5JKOQK *, +VQ ÷ þ2ýý1) +FGPVKHKECVKQP QH IGPGU TGSWKTGF HQT ITQYVJ WPFGT GVJCPQN UVTGUU WUKPI VTCPURQUQP OWVCIGPGUKU KP Saccharomyces cerevisiae. úQN ôGPGV ôGPQOKEU 2õó:1112-1119 6QJ *, *QTKOQVQ ÷ þ2ýý2) +PHGTGPEG QH C IGPGVKE PGVYQTM D[ C EQODKPGF CRRTQCEJ QH ENWUVGT CPCN[UKU CPF ITCRJKECN ôCWUUKCP OQFGNKPI. $KQKPHQTOCVKEU 18:287-297 6QPI #*, ©XCPIGNKUVC ú, öCTUQPU #$, ðW *, $CFGT ô&, öCIG 0, 4QDKPUQP ú, 4CIJKDK\CFGJ 5, *QIWG %ù, $WUUG[ *, #PFTGYU $, 6[GTU ú, $QQPG % þ2ýý1) 5[UVGOCVKE IGPGVKE CPCN[UKU YKVJ QTFGTGF CTTC[U QH [GCUV FGNGVKQP OWVCPVU. 5EKGPEG 29ü:23õü-23õ8 ñGV\ ö, ôKQV ï, %CIPG[ ô, úCPUHKGNF 6#, ÿWFUQP 45, ÷PKIJV ÿ4, ïQEMUJQP &, 0CTC[CP í, 5TKPKXCUCP ú, öQEJCTV ö, ìWTGUJK-©OKNK #, ïK ;, ôQFYKP $, %QPQXGT &, ÷CNDHNGKUEJ 6, íKLC[CFCOQFCT ô, ;CPI ú, ÿQJPUVQP ú, (KGNFU 5, 4QVJDGTI ÿú þ2ýýý) # EQORTGJGPUKXG CPCN[UKU QH RTQVGKP-RTQVGKP KPVGTCEVKQPU KP Saccharomyces cerevisiae. 0CVWTG üý3:õ23-õ27 XCP *GNFGP ÿ, #PFTG $, %QNNCFQ-íKFGU ÿ þ2ýýý) # YGD UKVG HQT VJG EQORWVCVKQPCN CPCN[UKU QH [GCUV TGIWNCVQT[ UGSWGPEGU. ;GCUV 1õ:177-187 íGNEWNGUEW í©, <JCPI ï, íQIGNUVGKP $, ÷KP\NGT ÷ù þ199ó) 5GTKCN CPCN[UKU QH IGPG GZRTGUUKQP. 5EKGPEG 27ý:ü8ü-ü87 íGNEWNGUEW í©, <JCPI ï, <JQW ù, íQIGNUVGKP ÿ, $CUTCK ú#, $CUUGVV &©, ÿT., *KGVGT ö, íQIGNUVGKP $, ÷KP\NGT ÷ù þ1997) %JCTCEVGTK\CVKQP QH VJG [GCUV VTCPUETKRVQOG. %GNN 88:2ü3-2ó1 ùCJNDQO (, ûVGTQ %, %QTFGTQ ûVGTQ 44, XCP <[N ù*, *CJP-*kIGTFCN $, ÿ{PUUQP ïÿ þ2ýý3) úQNGEWNCT CPCN[UKU QH C Saccharomyces cerevisiae OWVCPV YKVJ KORTQXGF CDKNKV[ VQ WVKNKUG Z[NQUG UJQYU GPJCPEGF GZRTGUUKQP QH RTQVGKPU KPXQNXGF KP VTCPURQTV, KPKVKCN Z[NQUG OGVCDQNKUO CPF VJG RGPVQUG RJQURJCVG RCVJYC[. #RRN ©PXKTQP úKETQDKQN õ9:7üý-7üõ ùCVGTUVQP 4*, GV CN. þ2ýý2) +PKVKCN UGSWGPEKPI CPF EQORCTCVKXG CPCN[UKU QH VJG OQWUG IGPQOG. 0CVWTG ü2ý:ó2ý-óõ2 ùGPFKUEJ í(, FG ôTCCH ##, 5CJO * þ1997) #EEWTCVG FGVGTOKPCVKQP QH 13% GPTKEJOGPVU KP PQPRTQVQPCVGF ECTDQP CVQOU QH KUQVQRQECNN[ GPTKEJGF COKPQ CEKFU D[ 1* PWENGCT OCIPGVKE TGUQPCPEG. #PCN $KQEJGO 2üó:19õ-2ý2 ùKGEJGTV ù þ2ýý1) 13% OGVCDQNKE HNWZ CPCN[UKU. úGVCD ©PI 3:19ó-2ýõ ùKP\GNGT ©#, 5JQGOCMGT &&, #UVTQOQHH #, ïKCPI *, #PFGTUQP ÷, #PFTG $, $CPIJCO 4, $GPKVQ 4, $QGMG ÿ&, $WUUG[ *, %JW #ú, %QPPGNN[ %, &CXKU ÷, &KGVTKEJ (, &QY 5ù, ©N $CMMQWT[ ú, (QWT[ (, (TKGPF 5*, ôGPVCNGP ©, ôKCGXGT ô, *GIGOCPP ÿ*, ÿQPGU 6, ïCWD ú, ïKCQ *, &CXKU 4ù þ1999) (WPEVKQPCN EJCTCEVGTK\CVKQP QH VJG S. cerevisiae IGPQOG D[ IGPG FGNGVKQP CPF RCTCNNGN CPCN[UKU. 5EKGPEG 28ó:9ý1-9ýõ
3õý
%JTKUVQHHGT $TQ, $KTIKVVG 4GIGPDGTI, CPF ÿGPU 0KGNUGP
ùKUGNQIGN #, 6[UQP 5, ÿQJPUQP & þ199õ) $KQOCUU HGGFUVQEM TGUQWTEGU CPF EQORQUKVKQP. +P: ù[OCP %© þGFU) *CPFDQQM QP DKQGVJCPQN: RTQFWEVKQP CPF WVKNK\CVKQP. 6C[NQT ê (TCPEKU, ùCUJKPIVQP, RR 1ýó-118 ùQFKEMC ï, &QPI *, úKVVOCPP ú, *Q ú*, ïQEMJCTV &ÿ þ1997) ôGPQOG-YKFG GZRTGUUKQP OQPKVQTKPI KP Saccharomyces cerevisiae. 0CV $KQVGEJPQN 1ó:13ó9-13õ7 ùQQF í, GV CN. þ2ýý2) 6JG IGPQOG UGSWGPEG QH Schizosaccharomyces pombe. 0CVWTG ü1ó:871-88ý
1ndex 2-de0xy-D-91uc05e, 75, 317 2-de0xy91uc05e, 152
A7F2, 154 A7H1, 77, 112
5,5,5-tr1f1u0r0-DL-1euc1ne, 154 5-f1u0r0-0r0t1c ac1d, 230 5-F0A 5e1ect10n, 84, 315
AUR1-C, 151
AAC2, 191
aux0tr0ph1c marker5, 35, 209, 307
A8C-pr0te1n5, 282 aceta1dehyde, 115 acet1c ac1d, 115 acety1-c0en2yme A, 184 A C51, 182 AC52, 182 act1ve dry yea5t, 101, 112 adeny1ate cyc1a5e, 64 ADH1, 183 AFLP, 102, 147 a1c0h01 0x1da5e, 208 ALD6, 127 ALK 9ene5, 250 a1kane5, 249 a110d1p101d, 149 a110p101d, 146 a1105ter1c c0ntr01, 338 a110tetrap101d, 149 Am1ne 0x1da5e, 211 am1ne5, 122 am1n0 ac1d accumu1at10n, 77 am1n0 ac1d5 6ranched-cha1n, 154
Ampe10myce5 4u154ua115,299
ant1free2e pr0te1n5, 82 ant1fun9a1 act1v1ty, 298 ant1fun9a1 yea5t5, 298
A0X1,210 A0X2, 210
a4uap0r1n5, 112 A4uap0r1n5, 82 AQY1, 82, 11~ AQY2, 82, 112 ara61n05e, 335 ar91n1ne, 114, 121 a5c05p0re5, 28 A7F1, 127, 147, 154
A7P, 113
aure06a51d1n A, 151
Aure06a51d1um pu11u1an5, 300
6aker•5 yea5t, 6, 29, 57 6ak1n9 perf0rmance, 58 8AP2, 146, 154 6eer 6rew1n9, 144 8EM1, 237 6ent0n1te, 116 6en201c ac1d, 276 86L2, 117 8HM1, 237 610c0ntr01, 11,297 6101nf0rmat1c5, 20, 285, 350 610ma55 y1e1d, 65 610pre5ervat10n, 123 6reed1n9, 30, 107, 148 6rewer•5 yea5t, 8, 144 6r0ad d0u9h ran9e, 39 8UD6, 237 6uty1 acetate, 302 ca1nex1n, 235 cAMP, 64 cAMP pathway, 239 CAN1, 146
Cand1da a161can5, 209, 236, 279 11p01yt1ca,275 01e0ph11a,298 CAR1, 77
car60hydrate meta60115m, 27 car60xypept1da5e Y, 218 CA78, 187 cata1a5e, 208 C8F1, 174 C8F2, 173 CDC25, 237 CDC42, 239 ce11 fact0ry, 208, 216, 332 ce11 512e, 84
362
1ndex
ce11 wa11, 277 ce11 wa115en50r, 235 CHEF, 308 ch1t1n, 280 ch1t1na5e, 300 CH51, 280 c1tr1c ac1d pr0duct10n, 227 c1ar1f1cat10n, 116 c1a551ca1 9enet1c5, 4 c01d 5tre55, 79 c0mp1ex 1, 240, 246 C0pper t0x1c1ty, 115 c0tran51at10na1 tran510cat10n, 232 C0X1, 176, 243 Cra6tree effect, 9, 172, 182 cry0pr0tect10n, 77 Cry0re515tance, 75
Crypt0c0ccu5 a161du5, 298 ne0f0rman5, 236, 279 C5H3, 280 C751, 123 C771, 83 CUP1, 115, 151
Cwp1, 283 CYC1, 146 CYC7, 146 CYR1, 64 cy5te1ne, 155 cyt0chr0me P450, 114, 250 cyt0duct10n, 30, 105 D1/D2 5e4uence5, 311
De6ary0myce5 han5en11, 308
D66E, 102 d1acety1, 157 d16en20furan, 249 d1methy15u1ph1de, 156 d1rected 6reed1n9, 149 DnaK, 234 d0m1nant marker, 150
ENA1, 73 END 9ene5, 214 ER610, 81
ethan01 t01erance, 114, 248 ethy1 car6amate, 121
FA51, 251 fatty ac1d
de5aturat10n, 81 fatty ac1d5, 81,114 F8A1, 79 F8P1, 75 fermentat10n perf0rmance, 111 fermentat1ve capac1ty, 58, 66 f1av0n01d5, 120 f1av0ur, 145, 154 f1av0ur 5ta6111ty, 151 FLD1, 212 FL01, 118, 159 FL010, 118 FL08, 118 f10ccu1at10n, 159 f10w cyt0metry, 32 f1ux0me, 346 FP51, 72, 125 free2e re515tance, 112 free2e re5p0n5e, 79 free2e-thaw re515tance, 26 free2e-thaw 5tre55, 84 fr02en d0u9h, 69, 76 F5Y1, 113 funct10na1 9en0m1c5, 4, 59, 130, 172, 285, 287, 332, 340 furan051da5e, 117 fu5e1 a1c0h015, 128 6 pr0te1n, 64 6418, 148, 210 6AL1, 188,337 6AL10, 188 6AL2, 113,337 6AL3, 189 6AL4, 337 6AL7, 188, 337 6AL80, 189, 337 9a1act05e, 337 9a551n9 p0wer, 58 9a551n9 rate, 58 6cr 1, 186 9ene c1u5ter ana1y515, 20, 23 c0nver510n5, 25 dup11cat10n, 20 expre5510n, 23 funct10n, 23 funct10na1 ana1y515, 22 0rder, 20
1ndex 9enet1c redtmdancy, 343 9en0m1c re0r9an12at10n, 60 6FP, 215 6LK1, 113 91ucan, 279 91ucan 5yntha5e, 277 91ucana5e, 116, 300 91uc0amy1a5e, 152 91uc0ne09ene515, 66 91uc05e, 63 repre5510n, 63, 152,186 tran5p0rt, 64 91uc05e 0x1da5e, 124 91utath10ne, 155 91ycer01, 70, 73, 127, 248 91yc09en, 112 91yc01y515,27, 66, 72, 177, 338 91yc01yt1cf1ux, 66, 113, 336 91yc05y1at10n, 283 91y0xy1ate cyc1e, 185 91y0xy1ate pathway, 249 6PD1, 72, 74, 125, 127 6PD2, 125, 127 6P1-anch0r, 282 6PR1, 249 6RA5, 31, 171, 192, 227 6rpE, 234 65H1, 83 65Y1, 112 65Y2, 112
HAL1, 73 ha10t01erant yea5t, 305 Han5enu1a p01ym0rpha, 11, 30, 207 HAP1, 187 HAP4, 66, 187 heter09ene1ty, 25 heter0tha111c, 28, 228, 306 heter02y9051ty, 34 hex05e tran5p0rter, 113, 179, 247, 280 H671, 180 h19h-9rav1ty fermentat10n, 152 H154, 146 H0, 147 H0 end0nuc1ea5e, 177 H 0 6 pathway, 74 h0m0e01090u5 chr0m050me5, 146 h0m01090u5 chr0m050me5, 146 h0m05er1ne, 155 h0m0tha111c, 28, 57
363
h05t defence, 298 H5P12, 74 H5P30, 118 H5p70, 234, 336 HXK1, 113 HXK2, 67, 113 HX79ene5, 64, 113 HX75, 280 hy6r1d, 146 hy6r1d 5tra1n5, 34 hy6r1d15at10n, 314 hy9r0myc1n 8, 230
1L V1, 146 1LV2, 147, 157 1L V3, 146 1LV5, 147 1ndu5tr1a1 fermentat10n, 335 1nver5e meta6011c en91neer1n9, 349 1nverta5e, 73 10n h0me05ta515, 75 10n1c e4u1116r1um, 73 150amy1 acetate, 154 175 re910n, 311 Kar2, 233 kary0type, 61 Kex2, 218 KEX2, 310 KH71, 179 KH72, 179 k111erp1a5m1d5, 175 k111ert0x1n, 114, 115, 124, 175, 191,298, 305,310 K10eckera ap1cu1ata, 309 K1uyver effect, 180 K1uyver0myce5 fra91115, 172 1act15, 9, 171 marx1anu5, 172 5pec1e5, 25
LAC 9ene5, 180 LAC12, 188 LAC4, 188 1act1c ac1d, 115 1act05e, 188 1a9er yea5t, 29, 146 LEA pr0te1n5, 74 LEU2, 146, 209
364
1ndex
LEU4, 154 L1ne e1ement5, 231 11p1d6105ynthe515, 27 L0RE, 286 LY51, 240 1y502yme, 123 macr0pex0pha9y, 213 MAL 10c1,40 MAL re9u1at0r, 42 ma101act1c fermentat10n, 123, 128 ma1ta5e, 44, 62, 67 ma1t05e, 40, 67, 152 ma1t05e permea5e, 44, 62, 67 ma1t05e ut1112at10n,44 ma1t0tr105e, 152 MAP-k1na5e, 287 ma55 mat1n9, 36 MA7, 146, 176, 229 me10t1c cr0551n9-0ver, 146 mem6rane f1u1d1ty, 76, 78, 81,285 5tructure, 75 Mende11an 5e9re9at10n, 18, 104 ME710, 147 ME714, 130 ME716, 130 ME72, 147 ME725, 155 ME73, 130 Meta6011c adju5tment, 72 meta6011c en91neer1n9, 11,332, 334 meta6011c f1ux, 109, 338 meta6011c m0de15, 350 meta6011c netw0rk, 346 meta6010me, 105, 346 methan01, 210 meth10n1ne, 155 Methy10tr0ph1c yea5t5, 208 m1cr0array ana1y515, 26, 81,342 m1cr0array, 23, 105 M1D2, 284 M191, 43, 67 M161, 337 m0de111n9, 347 m0n0car60xy11c ac1d5, 248 m0rph09ene515, 236 MRX1, 130 M5N1, 118 M5n2, 64
M5n4, 64 M5511,118 MUC1,118 muta9ene515, 30 MXR1,146,156 NADH, 72, 181 NADH dehydr09ena5e, 242 NADPH, 181 n1tr09en cata6011te repre5510n, 113 n1tr09en 11m1tat10n, 113 n0n-c0nvent10na1 yea5t5, 297 N0P1, 79 N7H1, 77 nutr1ent c0mpet1t10n, 302 0ff-f1av0ur5, 101,129, 145, 156 0LE1, 81,285 01eate, 217 05m0pr0tect10n, 72 05m0t1c 5tre55, 69 0x1dat1ve 5tre55, 83
PAD1, 129 pa5teur15at10n, 276 PCK1, 75 PDA1, 181 PDR 9ene5, 281 Pdr12, 278, 282 pect1n 1ya5e, 117 pect1na5e, 116 PEP4, 116 per0x150me a55em61y, 252, 254 per0x150me f0rmat10n, 213 per0x150me fu510n, 254 pet1te-ne9at1ve, 190 PEX 9ene5, 213,253 PEX19, 214 Pex20, 257 PEX3, 214 PEX8, 212 pex0pha9y, 214 PF6E, 311 PFK1, 180 PHD1, 237 phy109enet1c ana1y515,311 phyt0a1ex1ne5, 300 P1ch1a an0ma1a, 305 9u1111erm0nd11, 304 pa5t0r15, 11,207
1ndex
5t1p1t15,335 PKA, 64 Pkc1 pathway, 287 p1a5m1d 1nher1tance, 150 PMA1, 114 PMA2, 114 P0F1, 129 p01yp101d, 25, 103 p05t-tran51at10na1 m0d1f1cat10n, 208 pr0te0m1c 5, 27, 344 pre5ervat10n, 273 PR01, 78 PR02, 78 pr011ne, 78, 113 pr0m0ter fu510n5, 211 pr0p10ny1-C0A, 240 pr0tea5e, 116 pr0te1n ch1p5, 346 pr0te1n pr0duct10n, 228, 335 pr0te1n 5ecret10n, 218, 231 pr0te1n-DNA 1nteract10n5, 345 pr0te0me ana1y515, 105,339 pr0t0p1a5t fu510n, 314
P5eud02yma f10ccu105a, 300, 311
P75 519na1, 215 PU71, 78, 113 PU72, 114 putre5c1ne, 122
RAC1, 237
Ra9- phen0type, 179 179 183 180 180 183 185 Rap1, 174, 185 RAPD, 102,311 rare mat1n9, 150 Ra5, 64 RA52, 237 rat10na1 de519n, 332 rDNA 5trate91e5, 31 react1ve 0xy9en, 77 rec0m61nant techn0109y, 13 red0x 6a1ance, 70 REDUCE, 285, 342 re5p1rat0ry capac1ty, 66 re5p1rat0ry cha1n, 244
RA61, RA63, RA64, RA65, RA66, RA68,
re5veratr01, 120 RFLP, 102, 311 R672, 113
Rh0dut0ru1a 91ut1n15, 306 R05, 77, 83
R0X1, 187 RPL28, 79
5acchar0myce5, 19 6ayanu5, 148 car156er9en515, 26, 146 cerev151ae, 2, 100, 146 cerev151ae, d1a5tat1cu5, 153 k1uyver1, 151 1act15, 172 m0nacen515, 147 pa5t0r1anu5, 148 5en5u 5tr1ct0, 24 5pec1e5, 29
5acchar0myc0p515 11p01yt1ea, 228
5-aden05y1meth10n1ne, 155 5A6E, 342
5ch1205acehar0myce5 p0m6e, 3, 122, 128,341 5CR1, 232 5CR2, 232
5creen1n9, 38 5EC 9ene5, 230, 235 5EC61, 232 5EC62, 232 5EC65, 232 5ecret10n, 208 5ed1mentat10n, 159 5e1ect10n, 38 5en50ry 4ua11ty, 101,109, 125 5herry yea5t, 120 519na1 pept1da5e c0mp1ex, 218 519na1 rec09n1t10n pan1c1e, 232 519na1 tran5duct10n, 10, 284, 345 519na111n9pathway, 240 5KL-m0t1f, 216 5L51, 233 5L52, 234 51t2, 284 5MR1, 151 5NF1, 186 5n8, 180 5NF3, 113 50D1, 83
365
366
1ndex
50D2, 83 50K2, 237
50r61c ac1d, 276 50rt1n9, 212 5PC, 218 5pec1e5 5pec1f1c1ty, 173 5pher0p1a5t fu510n5, 37 5p0ru1at10n, 28, 62 5RP, 232 5RP101, 232 57E 9ene5, 239 5ter015, 112, 114, 277 5tra1n 1mpr0vement, 5 57RE, 286 5tre55 re515tance, 10, 39 re5p0n5e, 8, 27, 274, 282 t01erance, 112 5tuck fermentat10n5, 113, 115 5u1phate a551m11at10n, 157 5u1ph1te, 114, 115, 129, 155 5u1ph1te eff1ux, 157 5uper0x1de d15muta5e, 83 5U71, 112 5U72, 112 5ynthet1c 1etha1 5creen, 233 tar9et f1nd1n9, 347 tar9et1n9 519na15,212 te1105p0re5, 306 terpen01d5, 126 thaw1n9, 76, 77 71P1, 79 7 0 R pathway, 73
70ru1a5p0ra de16rueck11, 9, 83 7P11, 81, 181 7P51, 112 7P52, 112
tran5cr1pt0me, 160, 339 tran5cr1pt0m1c5, 27, 284 tran5p050n muta9ene515, 230 treha105e, 77, 112 treha105e-6-ph05phate, 112, 247 7RX2, 83 7UP1, 238 tw0-hy6r1d a55ay, 23,345 7y e1ement5, 25 7y1 tran5p050n, 172 u614u1n0ne, 242
Unf01ded Pr0te1n Re5p0n5e, 235 UPR, 235 URA3, 146, 209
URE2, 113
urea, 121
U5t11a90 mayd15, 236 VAR1, 176 VP5 9ene5, 214
weak ac1d 5tre55, 282 weak 0r9an1c ac1d5, 276 w1ne fermentat10n, 101 w1ne yea5t, 8, 27, 74, 100 W5C 9ene5, 284 W5C1, 284 xy1ana5e, 117 xy105e, 335,349 xy1051da5e, 117
YAP1, 83 Yarr0w1a 11p01yt1ca,227
yea5t 610d1ver51ty, 2 610techn0109y, 3,298 f10ccu1at10n, 118 f10tat10n, 118 f00d 5p011a9e, 273 9enet1c5, 18 9en0me, 22 phy510109y, 3, 109 yea5t 610techn0109y, 192 yea5t ec0109y, 273 yea5t phy510109y, 177 YHC8, 234 YP71, 212 2eta re910n, 230
2y905acchar0myce5 6a1111,273 615p0ru5, 276 1entu5, 276 r0ux11, 26, 275,305
2ym0c1n, 115, 123, 175, 191 2ym01ya5e, 283 a-acet01actate, 157
1ndex
1~-0x1dat10n, 253
~,-deca1act0ne, 251
367
