Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Expression and Localization of Inulinase in Recombinant Saccharomyces cerevisiae

재조합 Saccharomyces cerevisiae에서 Inulinase의 발현과 국재성

  • 남수완 (한국과학기술연구원 유전공학연구소) ;
  • 우문희 (한국과학기술연구원 유전공학연구소) ;
  • 김병문 (한국과학기술연구원 유전공학연구소) ;
  • 정봉현 (한국과학기술연구원 유전공학연구소) ;
  • 박영훈 (한국과학기술연구원 유전공학연구소)
  • Published : 1994.04.01
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Abstract

Inulinase of Kluyveromyces marxianus origin was produced by recombinant yeast Saccharomyces cerevisiae under the control of GAL1 promoter, to examine the expression and localization of inulinase in a repressed(galactose-free) or derepressed(galactose-containinga) medium. The inulinase gene(INU1A) was constitutively expressed at 6.7 units/ml in a repressed medium. When the cell started to utilize galactose in a derepressed medium, the INU1A gene began to be expressed, and the final expression level reached about 45 units/ml. According to be the nondenaturingPAGE analysis, inulinase produced by S. cerevisiae was found to be less glycosylated than the bakers yeast invertase. In addition, its glycosylation pattern was less heterogeneous than the K. marxianus inulinase. The supplementation of inulin or raffinose into the derepressed medium increased the cell growth rate, while the expression of INU1A was repressed. Regardless of the carbon sources examined, most of inulinase activity (more than 98%) was found in the extracellular medium, indicating excellent secretion efficiency.

Keywords

References

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