期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 69, 期 13, 页码 3887-3894出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.1c00158
关键词
SpyCatcher/SpyTag; multienzyme systems; nanoaggregates; reusability; glutathione
资金
- China National Key Research and Development Program [2019YFA0904300]
- Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project [TSBICIP-KJGG-009]
- Fundamental Research Funds for the Central Universities [22221818014]
The study established a system that can assemble into multienzyme systems to enhance the production of glutathione and oxidized glutathione, with only a 9.4% decrease in total yield after five continuous rounds of biocatalysis.
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.
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