Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Creation of an Ethanol-Tolerant Yeast Strain by Genome Reconstruction Based on Chromosome Splitting Technology

  • Park, A-Hwang (Department of Biomaterial Control (BK 21 Program), Dong-Eui University) ;
  • Sugiyama, Minetaka (Department of Biotechnology, Graduate School of Engineering, Osaka University) ;
  • Harashima, Satoshi (Department of Biotechnology, Graduate School of Engineering, Osaka University) ;
  • Kim, Yeon-Hee (Department of Biomaterial Control (BK 21 Program), Dong-Eui University)
  • Received : 2011.09.19
  • Accepted : 2011.11.08
  • Published : 2012.02.28
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Abstract

We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.

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References

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