Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae

Z. Petek Çakar; Urartu O.S. Seker; Candan Tamerler; Marco Sonderegger; Uwe Sauer

doi:10.1016/j.femsyr.2004.10.010

Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae

Z. Petek Çakar^*, Urartu O.S. Seker, Candan Tamerler, Marco Sonderegger, Uwe Sauer

^*Corresponding author for this work

Department of Molecular Biology and Genetics

Research output: Contribution to journal › Article › peer-review

163 Citations (Scopus)

Abstract

Various selection procedures in chemostats and batch cultures were systematically tested for their efficiency to select for a multiple-stress resistance phenotype in Saccharomyces cerevisiae. To determine the relative stress resistance phenotypes, mutant populations harvested at different time points and randomly chosen clones from selected populations were grown in batch cultures and exposed to oxidative, freezing-thawing, high-temperature and ethanol stress. For this purpose, we developed a high-throughput procedure in 96-well plates combined with a most-probable-number assay. Among all chemostat and batch selection strategies tested, the best selection strategy to obtain highly improved multiple-stress-resistant yeast was found to be batch selection for freezing-thawing stress. The final mutant populations selected for this particular stress were not only significantly improved in freezing-thawing stress resistance, but also in other stress resistances. The best isolated clone from these populations exhibited 102-, 89-, 62-, and 1429-fold increased resistance to freezing-thawing, temperature, ethanol, and oxidative stress, respectively. General selection guidelines for improving multiple-stress resistance in S. cerevisiae are presented and discussed.

Original language	English
Pages (from-to)	569-578
Number of pages	10
Journal	FEMS Yeast Research
Volume	5
Issue number	6-7
DOIs	https://doi.org/10.1016/j.femsyr.2004.10.010
Publication status	Published - Apr 2005

Funding

We are grateful to James E. Bailey for the initial support. We thank Hande Akçe, Tugba Aloglu, Marcel Emmerling and Peter Steiner for technical assistance. This work was supported by Istanbul Technical University (ITU) Research Funds (PI: Z.P. Çakar, Project No: 30108), Turkish State Planning Organization (DPT) (PI: G. Saglamer) and the Swiss Bundesamt fuer Bildung und Wissenschaft (BBW) within the Framework IV Biotechnology Programme of the European Commission.

Funders	Funder number
BBW
DPT
Turkish State Planning Organization
European Commission
Istanbul Teknik Üniversitesi	30108
Office Fédéral de l'Education et de la Science

Keywords

Evolution
Freezing stress
Inverse metabolic engineering
Selection
Stress resistance

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10.1016/j.femsyr.2004.10.010

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title = "Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae",

abstract = "Various selection procedures in chemostats and batch cultures were systematically tested for their efficiency to select for a multiple-stress resistance phenotype in Saccharomyces cerevisiae. To determine the relative stress resistance phenotypes, mutant populations harvested at different time points and randomly chosen clones from selected populations were grown in batch cultures and exposed to oxidative, freezing-thawing, high-temperature and ethanol stress. For this purpose, we developed a high-throughput procedure in 96-well plates combined with a most-probable-number assay. Among all chemostat and batch selection strategies tested, the best selection strategy to obtain highly improved multiple-stress-resistant yeast was found to be batch selection for freezing-thawing stress. The final mutant populations selected for this particular stress were not only significantly improved in freezing-thawing stress resistance, but also in other stress resistances. The best isolated clone from these populations exhibited 102-, 89-, 62-, and 1429-fold increased resistance to freezing-thawing, temperature, ethanol, and oxidative stress, respectively. General selection guidelines for improving multiple-stress resistance in S. cerevisiae are presented and discussed.",

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AU - Tamerler, Candan

AU - Sonderegger, Marco

AU - Sauer, Uwe

PY - 2005/4

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Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae

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Keywords

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