Genome shuffling to improve thermotolerance,ethanol tolerance and ethanol productivity of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> |
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Authors: | Dong-jian Shi Chang-lu Wang Kui-ming Wang |
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Institution: | (1) Laboratory of Food Biotechnology, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, 300457 Tianjin, People’s Republic of China |
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Abstract: | Genome shuffling is a powerful strategy for rapid engineering of microbial strains for desirable industrial phenotypes. Here
we improved the thermotolerance and ethanol tolerance of an industrial yeast strain SM-3 by genome shuffling while simultaneously
enhancing the ethanol productivity. The starting population was generated by protoplast ultraviolet irradiation and then subjected
for the recursive protoplast fusion. The positive colonies from the library, created by fusing the inactivated protoplasts
were screened for growth at 35, 40, 45, 50 and 55°C on YPD-agar plates containing different concentrations of ethanol. Characterization
of all mutants and wild-type strain in the shake-flask indicated the compatibility of three phenotypes of thermotolerance,
ethanol tolerance and ethanol yields enhancement. After three rounds of genome shuffling, the best performing strain, F34,
which could grow on plate cultures up to 55°C, was obtained. It was found capable of completely utilizing 20% (w/v) glucose
at 45–48°C, producing 9.95% (w/v) ethanol, and tolerating 25% (v/v) ethanol stress. |
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Keywords: | Ethanol production Genome shuffling Saccharomyces cerevisiae Thermotolerance |
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