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Generation and characterisation of stable ethanol-tolerant mutants of Saccharomyces cerevisiae

Journal of Industrial Microbiology & Biotechnology volume 37, pages 139–149 (2010)Cite this article

Abstract

Saccharomyces spp. are widely used for ethanologenic fermentations, however yeast metabolic rate and viability decrease as ethanol accumulates during fermentation, compromising ethanol yield. Improving ethanol tolerance in yeast should, therefore, reduce the impact of ethanol toxicity on fermentation performance. The purpose of the current work was to generate and characterise ethanol-tolerant yeast mutants by subjecting mutagenised and non-mutagenised populations of Saccharomyces cerevisiae W303-1A to adaptive evolution using ethanol stress as a selection pressure. Mutants CM1 (chemically mutagenised) and SM1 (spontaneous) had increased acclimation and growth rates when cultivated in sub-lethal ethanol concentrations, and their survivability in lethal ethanol concentrations was considerably improved compared with the parent strain. The mutants utilised glucose at a higher rate than the parent in the presence of ethanol and an initial glucose concentration of 20 g l−1. At a glucose concentration of 100 g l−1, SM1 had the highest glucose utilisation rate in the presence or absence of ethanol. The mutants produced substantially more glycerol than the parent and, although acetate was only detectable in ethanol-stressed cultures, both mutants produced more acetate than the parent. It is suggested that the increased ethanol tolerance of the mutants is due to their elevated glycerol production rates and the potential of this to increase the ratio of oxidised and reduced forms of nicotinamide adenine dinucleotide (NAD+/NADH) in an ethanol-compromised cell, stimulating glycolytic activity.

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Acknowledgments

The authors thank Foster’s Group Ltd. and The Australian Wine Research Institute for project support. The authors also thank the Australian Government for providing a Commonwealth Postgraduate Award to support Dragana Stanley.

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Authors and Affiliations

  1. School of Engineering and Science, Victoria University, PO Box 14428, Melbourne, VIC, 8001, Australia

    Dragana Stanley, Sarah Fraser & Grant A. Stanley

  2. The Australian Wine Research Institute, PO Box 197, Glen Osmond, SA, 5064, Australia

    Paul J. Chambers

  3. Foster’s Group Ltd, 4 Southampton Crescent, Abbotsford, VIC, 3067, Australia

    Peter Rogers

Authors
  1. Dragana Stanley
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  2. Sarah Fraser
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  3. Paul J. Chambers
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  4. Peter Rogers
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  5. Grant A. Stanley
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Correspondence to Grant A. Stanley.

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Stanley, D., Fraser, S., Chambers, P.J. et al. Generation and characterisation of stable ethanol-tolerant mutants of Saccharomyces cerevisiae . J Ind Microbiol Biotechnol 37, 139–149 (2010). https://doi.org/10.1007/s10295-009-0655-3

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  • DOI: https://doi.org/10.1007/s10295-009-0655-3

Keywords

  • Saccharomyces
  • Ethanol
  • Stress
  • Mutant
  • NADH