Abstract
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.
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This work was supported by the National Basic Research Program of China (973: 2014CB745101), Knowledge Innovation Program (KSZD-EW-Z-019), and Science and Technology Service Network Initiative (KFJ-EW-STS-030) of the Chinese Academy of Sciences, and Shanghai Scientific research project (14XD1424900).
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Jiang, Y., Tao, R., Shen, Z. et al. Enzymatic Production of Glutathione by Bifunctional γ-Glutamylcysteine Synthetase/Glutathione Synthetase Coupled with In Vitro Acetate Kinase-Based ATP Generation. Appl Biochem Biotechnol 180, 1446–1455 (2016). https://doi.org/10.1007/s12010-016-2178-5
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DOI: https://doi.org/10.1007/s12010-016-2178-5