In-vitro Production of Glutathione Using Yeast ATP Regeneration System and Recombinant Synthetic Enzymes from Escherichia coli.

효모의 ATP 재생산계와 대장균 유래의 재조합 생산효소를 이용한 in vitro 글루타치온 생산

  • 고성영 (인하대학교 공과대학 생물공학과) ;
  • 구윤모 (인하대학교 공과대학 생물공학과)
  • Published : 1998.06.01

Abstract

An ATP regeneration system was used for the production of glutathione which was synthesized by a sequential action of ${\gamma}$-glutamyl-cysteine synthetase and glutathione synthetase. The synthetases above were produced in the recombinant E. coli (TG1/pDG7) with the highest specific production yield of 31 mg glutathione/g wet cell. Bakers yeast was considered to have economically a better ATP regeneration system although the glutathione production yield was lower than that of acetate kinase. It was also observed that the ATP regeneration system of bakers yeast was superior to that of Saccharomyces cerevisiae ATCC24858. The yield of glutathione production with bakers yeast was 36% with the ATP concentration of 5 mM. To avoid the cysteine limitation during the early phase of glutatione production, an extra cysteine was added at 2 hours after reaction and the production yield increased 1.91 times. The effectiveness of bakers yeast as an ATP regeneration system was proved by several sets of extra feeding experiments. The product inhibition by glutathione above 14 mM was also observed.