文献类型：期刊文献

英文题名：LProgramming Integrative Multienzyme Systems and Ionic Strength For Recyclable Synthesis of Glutathione

作者：Cui, Xiangwei[1];Wang, Zeyuan[1];Li, Zonglin[1];Zhang, Xing[1];Li, Zhimin[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg Technol, Shanghai 200237, Peoples R China

年份：2021

卷号：69

期号：13

起止页码：3887

外文期刊名：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000639018000009)】；

基金：This study was supported by the China National Key Research and Development Program (Grant No. 2019YFA0904300.), the Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (Grant No. TSBICIP-KJGG-009), and the Fundamental Research Funds for the Central Universities (22221818014).

语种：英文

外文关键词：SpyCatcher/SpyTag; multienzyme systems; nanoaggregates; reusability; glutathione

摘要：In the enzymatic cascade catalysis, it is a big challenge to construct a stable and reusable catalyst with targeted enzymes. The artificial multienzyme reactor has attracted great attention due to its potential for facilitating the performance of enzyme catalysis. In this study, we set up a reliable system that could assemble polyphosphate kinase (PPK) with bifunctional glutathione synthetase (GshF) via SpyCatcher/SpyTag to form multienzyme systems (MESs). Furthermore, MESs could assemble into nanoaggregates by altering the ionic strength, and the larger nanoaggregates could be applied in robust and reusable synthesis of glutathione (GSH). To enhance MES levels in vivo, gene duplication and different coexpression modes were performed. Finally, the optimized production of GSH and oxidized glutathione (GSSG) reached 102.6 and 6.7 mM within 2 h. Compared with the first round, the total yield only decreased by 9.4% after five continuous rounds of biocatalysis.