Enzymatic Production of Glutathione by Bifunctional γ-Glutamylcysteine Synthetase/Glutathione Synthetase Coupled with In Vitro Acetate Kinase-Based ATP Generation

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• 作者：Yu Jiang ; Rongsheng Tao ; Zhengquan Shen…
• 关键词：Acetate kinase ; ATP generation ; Bifunctional γ ; glutamylcysteine synthetase/glutathione synthetase ; Glutathione
• 刊名：Applied Biochemistry and Biotechnology
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：180
• 期：7
• 页码：1446-1455
• 全文大小：
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Biotechnology; Biochemistry, general;
• 出版者：Springer US
• ISSN：1559-0291
• 卷排序：180

文摘

Glutathione (γ-glutamyl-L-cysteinylglycine, GSH) is a pharmaceutical compound often used in food additives and the cosmetics industry. GSH can be produced biologically from L-glutamic acid, L-cysteine, and glycine through an enzymatic process traditionally involving two sequential adenosine triphosphate (ATP)-dependent reactions catalyzed by γ-glutamylcysteine synthetase (γ-GCS or GSHI, EC 6.3.2.2) and GSH synthetase (GS or GSHII, EC 6.3.2.3). Here, we report the enzymatic production of GSH by recombinant cell-free bifunctional γ-glutamylcysteine synthetase/glutathione synthetase (γ-GCS-GS or GshF) coupled with in vitro acetate kinase-based ATP generation. GSH production by an acetate kinase-integrated Escherichia coli Rosetta(DE3) mutant expressing Streptococcus thermophilus GshF reached 18.3 ± 0.1 g l−1 (59.5 ± 0.3 mM) within 3 h, with a molar yield of 0.75 ± 0.00 mol mol−1 added cysteine and a productivity of 6.1 ± 0.0 g l−1 h−1. This is the highest GSH titer reported to date. This newly developed biocatalytic process offers a promising approach for meeting the industrial requirements for GSH production.