Efficient production of l-glutathione by whole-cell catalysis with ATP regeneration from adenosine

Siqi Zuo; Jiarun Zhao; Pengfei Zhang; Wenqian Liu; Ke Bi; Peilian Wei; Jiazhang Lian; Zhinan Xu

doi:10.1002/bit.28711

Efficient production of l-glutathione by whole-cell catalysis with ATP regeneration from adenosine

Biotechnol Bioeng. 2024 Apr 17. doi: 10.1002/bit.28711. Online ahead of print.

Authors

Siqi Zuo^{1

2}, Jiarun Zhao^{1

2}, Pengfei Zhang³, Wenqian Liu^{1

2}, Ke Bi^{1

2}, Peilian Wei³, Jiazhang Lian^{1

2}, Zhinan Xu^{1

2}

Affiliations

¹ Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
² Institute of Biological Engineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
³ School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou, China.

PMID: 38629468
DOI: 10.1002/bit.28711

Abstract

l-glutathione (GSH) is an important tripeptide compound with extensive applications in medicine, food additives, and cosmetics industries. In this work, an innovative whole-cell catalytic strategy was developed to enhance GSH production by combining metabolic engineering of GSH biosynthetic pathways with an adenosine-based adenosine triphosphate (ATP) regeneration system in Escherichia coli. Concretely, to enhance GSH production in E. coli, several genes associated with GSH and l-cysteine degradation, as well as the branched metabolic flow, were deleted. Additionally, the GSH bifunctional synthase (GshFSA) and GSH ATP-binding cassette exporter (CydDC) were overexpressed. Moreover, an adenosine-based ATP regeneration system was first introduced into E. coli to enhance GSH biosynthesis without exogenous ATP additions. Through the optimization of whole-cell catalytic conditions, the engineered strain GSH17-FDC achieved an impressive GSH titer of 24.19 g/L only after 2 h reaction, with a nearly 100% (98.39%) conversion rate from the added l-Cys. This work not only unveils a new platform for GSH production but also provides valuable insights for the production of other high-value metabolites that rely on ATP consumption.

Keywords: ATP regeneration; GshF; adenosine; l‐glutathione; whole‐cell catalysis.

Abstract

Grants and funding