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Computer-Aided Semi-Rational Design to Enhance the Activity of l-Sorbosone Dehydrogenase from Gluconobacter oxidans WSH-004

J Agric Food Chem. 2024-05; 
Dong Li, Xinglong Wang, Lin Huo, Weizhu Zeng, Jianghua Li, Jingwen Zhou
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Codon Optimization The SNDH gene was optimized by GenScript Biotech Corporation and expressed in E. coli BL21(DE3) expressing L-sorbose dehydrogenase18 using the pET28a- (+) plasmid to verify the activity of SNDH and its mutants. Get A Quote

摘要

The titer of the microbial fermentation products can be increased by enzyme engineering. l-Sorbosone dehydrogenase (SNDH) is a key enzyme in the production of 2-keto-l-gulonic acid (2-KLG), which is the precursor of vitamin C. Enhancing the activity of SNDH may have a positive impact on 2-KLG production. In this study, a computer-aided semirational design of SNDH was conducted. Based on the analysis of SNDH's substrate pocket and multiple sequence alignment, three modification strategies were established: (1) expanding the entrance of SNDH's substrate pocket, (2) engineering the residues within the substrate pocket, and (3) enhancing the electron transfer of SNDH. Finally, mutants S453A, L460V, and E471D were o... More

关键词

2-keto-l-gulonic acid; computer-aided simulation design; l-sorbosone dehydrogenase; semirational design; substrate pocket.