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Construction, expression, purification, characterization, and structural analysis of microbial transglutaminase variants

Biotechnol Appl Biochem. 2021-12; 
Xiaoping Song, Hefang Sheng, Yueqiao Zhou, Yin Yu, Yingjiao He, Zihan Wang
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Recombinant Antibody Expression … Recombinant Protein Pharmaceutical Biotechnology, Hefei 230022, China *Corresponding author … The gene (mtg1) of Mut1 was synthesized by GenScript Biotech Co., LTD … Get A Quote

摘要

Microbial transglutaminase (MTG, EC 2.3.2.13) derived from Streptomyces mobaraensis is widely used in the food and pharmaceutical industry because of its ability to synthesize isopeptide bonds between the proteinogenic side chains of glutamine and lysine. The half-life (t ) of the activated wild-type enzyme at 60°C is 2 min. To improve the activity and thermostability of MTG for higher temperature application, three variants (Mut1, Mut2, and Mut3) were obtained by combining key amino acid mutations on the basis of previous research results. The best variant Mut2 with a specific combination of five of seven substitutions (S2P-S23V-Y24N-R215A-H289Y) shows a 10-fold increased half-life at 60°C (t  = 27.6 min... More

关键词

Raman spectra, circular dichroism, combination of mutations, microbial transglutaminase, specific enzyme activity, thermostability