Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Overexpression of Thermoalkalophilic Lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae

  • Ahn, Jung-Oh (Department of Chemical Engineering, Yonsei University) ;
  • Jang, Hyung-Wook (Acebiotech Corporation, Chungwon) ;
  • Lee, Hong-Weon (Bioventure Center, Bio-pilot Plant, Korea Research Institute of Bioscience and Biotechnology) ;
  • Choi, Eui-Sung (Microbial Functions Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • Haam, Seung-Joo (Department of Chemical Engineering, Yonsei University) ;
  • Oh, Tae-Kwang (Microbial Genomics Applications Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Jung, Joon-Ki (Bioventure Center, Bio-pilot Plant, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2003.06.01
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Abstract

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

Keywords

References

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