Abstract
Saccharomyces cerevisiae was transformed for higher ethanol tolerance by error-prone whole genome amplification. The resulting PCR products were transformed back to the parental strain for homologous recombination to create a library of mutants with the perturbed genomic networks. A few rounds of transformation led to the isolation of mutants that grew in 9% (v/v) ethanol and 100 g glucose l−1 compared to untransformed yeast which grew only at 6% (v/v) ethanol and 100 g glucose l−1.
Similar content being viewed by others
References
Alper H, Moxley J, Nevoigt E, Fink GR, Stephanopoulos G (2006) Engineering yeast transcription machinery for improved ethanol tolerance and production. Science 314:1565–1568
Crameri A, Raillard SA, Bermudez E, Stemmer WP (1998) DNA shuffling of a family of genes from diverse species accelerates directed evolution. Nature 39:288–291
Lee KJ, Tribe DE, Rogers PL (1979) Ethanol production by Zymomonas mobilis in continuous culture at high glucose concentration. Biotechnol Lett 1:421–426
Patnaik R (2008) Engineering complex phenotypes in industrial strains. Biotechnol Prog 24:38–47
Stouthamer AH, Bettenhaussen C (1973) Utilization of energy for growth and maintenance in continuous and batch cultures of microorganisms. A reevaluation of the method for the determination of ATP production by measuring molar growth yields. Biochim Biophys Acta 301:53–70
Xu B, Jin Z, Wang H, Jin Q, Jin X, Cen P (2008) Evolution of Streptomyces pristinaespiralis for resistance and production of pristinamycin by genome shuffling. Appl Microbiol Biotechnol 80:261–267
Zhang L, Cui X, Schmitt K, Hubert R, Navidi W (1992) Whole genome amplification from a single cell: implications for genetic analysis. Proc Natl Acad Sci USA 89:5847–5851
Acknowledgments
We thank the Industrial Biotechnology Group members for their feedback and suggestions. This work was funded by Agency for Science, Technology and Research (A*STAR) in Singapore ICES/07-173B01.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Luhe, A.L., Tan, L., Wu, J. et al. Increase of ethanol tolerance of Saccharomyces cerevisiae by error-prone whole genome amplification. Biotechnol Lett 33, 1007–1011 (2011). https://doi.org/10.1007/s10529-011-0518-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-011-0518-7