Abstract
Two single gene cassettes, each containing one of the individual gene (γ-glutamylcysteine synthetase gene GSH1 or glutathione synthetase gene GSH2), were constructed under the control of alcohol dehydrogenase (ADH1) promoter and their respective native terminators. The recombinant plasmids constructed with Kan r or Hyg r as the selective markers and were transformed into Saccharomyces cerevisiae separately and jointly. Three engineered strains, GSH1-enhanced strain S.TS013/GSH1, GSH2-enhanced strain S.TS013/GSH2 and GSH1+GSH2 double-enhanced strain S.TS013/GSH1+GSH2, were constructed. Glutathione production using the recombinant strains to improve was then determined. By the cell dosage proportions of two engineered strains (S.TS013/GSH1, S.TS013/GSH2) and a two-stage reaction, GSH productivity increased by 84 and 59 % over that of the host strain and the S.TS013/GSH1+GSH2 strain, respectively.
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Acknowledgments
We are deeply grateful to Prof. Guojun Huang (Nanchang University, China) for providing the YEplace181, pFA6a-kanMX6, pYM20 plasmids. This study was supported by grant from State Key Laboratory of Food Science and Technology, Nanchang University (SKLFTS), No. 201113. All the authors declare no interest conflict in this study.
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Chen, Jl., Xie, L., Cai, Jj. et al. Enzymatic synthesis of glutathione using engineered Saccharomyces cerevisiae . Biotechnol Lett 35, 1259–1264 (2013). https://doi.org/10.1007/s10529-013-1191-9
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DOI: https://doi.org/10.1007/s10529-013-1191-9