Thermus caldophilus GK24로부터 내열성 $\beta$-galactosidase의 최적 생산

  • 유진상 (성균관대학교 유전공학과) ;
  • 김현규 (성균관대학교 유전공학과) ;
  • 인만진 ((주)미원 중앙연구소) ;
  • 김민홍 ((주)미원 중앙연구소) ;
  • 권석태 (성균관대학교 유전공학과)
  • Published : 1997.06.01

Abstract

Thermus caldophilus GK24 was selected as sources of thermostable $\beta$-galactosidase from a survey of genus Thermus. T. caldophilus GK24 (Tca) $\beta$-galactosidase was found to be inducible. The enzyme was optimally active at 75$\circ$C. Enzyme induction was achieved by addition of lactose, galactose and cellobiose to basal media. The addition of glucose to culture media had a repressive effect on further enzyme synthesis. T caldophilus GK24 was tested for production of $\beta$-galactosidase by addition of various concentration of lactose, galactose and cellobiose to standard media. Cellobiose was found to be effective for the $\beta$-galactosidase induction. The optimal induction medium for production of $\beta$-galactosidase was composed of 0.2% cellobiose, 0.3% bactotryptone, 0.3% yeast extract, basal salts and Tris/HCI(pH 7.8). The activity of the enzyme in the optimal induction medium increased nearly 16.5-fold compared to the standard medium. Tca $\beta$-galactosidase was detected when cell extracts was subjected to electrophoresis in a nondenaturing polyacryamide gel and stained for activity with 6-bromo-2-naphtyl-$\beta$-D-galactopyranoside(BNG).

Keywords

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