Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae. 2005

Z Petek Cakar, and Urartu O S Seker, and Candan Tamerler, and Marco Sonderegger, and Uwe Sauer
Department of Molecular Biology and Genetics, Faculty of Science & Letters, Istanbul Technical University, Maslak, TR-34469 Istanbul, Turkey. cakarp@itu.edu.tr

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.

UI MeSH Term Description Entries
D009154 Mutation Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations. Mutations
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D005818 Genetic Engineering Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc. Genetic Intervention,Engineering, Genetic,Intervention, Genetic,Genetic Interventions,Interventions, Genetic
D000431 Ethanol A clear, colorless liquid rapidly absorbed from the gastrointestinal tract and distributed throughout the body. It has bactericidal activity and is used often as a topical disinfectant. It is widely used as a solvent and preservative in pharmaceutical preparations as well as serving as the primary ingredient in ALCOHOLIC BEVERAGES. Alcohol, Ethyl,Absolute Alcohol,Grain Alcohol,Alcohol, Absolute,Alcohol, Grain,Ethyl Alcohol
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D012641 Selection, Genetic Differential and non-random reproduction of different genotypes, operating to alter the gene frequencies within a population. Natural Selection,Genetic Selection,Selection, Natural
D018869 Heat-Shock Response A sequence of responses that occur when an organism is exposed to excessive heat. In humans, an increase in skin temperature triggers muscle relaxation, sweating, and vasodilation. Heat-Shock Reaction,Heat Shock,Heat Shock Stress,Heat Stress,Heat-Stress Reaction,Heat-Stress Response,Heat Shock Reaction,Heat Shock Response,Heat Shock Stresses,Heat Shocks,Heat Stress Reaction,Heat Stress Response,Heat Stresses,Heat-Shock Reactions,Heat-Shock Responses,Heat-Stress Reactions,Heat-Stress Responses,Shock, Heat,Stress, Heat,Stress, Heat Shock
D019020 Directed Molecular Evolution The techniques used to produce molecules exhibiting properties that conform to the demands of the experimenter. These techniques combine methods of generating structural changes with methods of selection. They are also used to examine proposed mechanisms of evolution under in vitro selection conditions. Evolution, Molecular, Directed,Molecular Evolution, Directed,In Vitro Molecular Evolution,Laboratory Molecular Evolution,Directed Molecular Evolutions,Evolution, Directed Molecular,Evolution, Laboratory Molecular,Evolutions, Directed Molecular,Evolutions, Laboratory Molecular,Laboratory Molecular Evolutions,Molecular Evolution, Laboratory,Molecular Evolutions, Directed,Molecular Evolutions, Laboratory

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