文献类型：期刊文献

英文题名：Production of glutathione using a bifunctional enzyme encoded by gshF from Streptococcus thermophilus expressed in Escherichia coli

作者：Li, Wei[1];Li, Zhimin[1];Yang, Jianhua[1];Ye, Qin[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2011

卷号：154

期号：4

起止页码：261

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000292958600011)】；

基金：We are deeply grateful to Prof. Shuang Li (SIIM, Shanghai, China) for providing the strain S. thermophilus SIIM B218, and Prof. Satoshi Harashima (Osaka University, Japan) for providing the mutant strains of S. cerevisiae. This study was supported by the National Special Fund for State Key Laboratory of Bioreactor Engineering, Grant No. 2060204, and the project of Shanghai Leading Academic Disciplines (No. B505).

语种：英文

外文关键词：Glutathione; Bifunctional glutathione synthetase; Feedback inhibition; ATP regeneration system

摘要：Glutathione (GSH) is one of the most ubiquitous non-protein thiols that is involved in numerous cellular activities. The gene coding for a novel bifunctional enzyme catalyzing the reaction for glutathione synthesis, gshF, was cloned from Streptococcus thermophilus SIIM B218 and expressed in Escherichia coli JM109. In the presence of the precursor amino acids and ATP, the induced cells of E. coli JM109 (pTrc99A-gshF) could accumulate 10.3 mM GSH in 5 h. The S. thermophilus GshF was insensitive to feedback inhibition caused by GSH even at 20 mM. At elevated concentrations of the precursor amino acids and ATP, E. coli JM109 (pTrc99A-gshF) produced 36 mM GSH with a molar yield of 0.9 mol/mol based on added cysteine and of 0.45 mol/mol based on added ATP. When ATP was replaced with glucose, E. coli JM109 (pTrc99A-gshF) produced 7 mM in 3 h. Saccharomyces cerevisiae was used to generate ATP for GSH production. In the presence of glucose and the pmr1 mutant of S. cerevisiae BY4742, JM109 (pTrc99A-gshF) produced 33.9 mM GSH in 12 h with a yield of 0.85 mol/mol based on added l-cysteine. It is shown that the S. thermophilus GshF can be successfully used for GSH production. (C) 2011 Elsevier B. V. All rights reserved.