Characterization of a new L-carnosine synthase mined from deep-sea sediment metagenome

Microbial Cell Factories - Tập 21 - Trang 1-10 - 2022
Jiajia She1,2, Lihong Fu1, Xiaowei Zheng2, Jing Li1,2, Limin Wang2, Bo Yu2, Jiansong Ju1,3
1College of Life Science, Hebei Normal University, Shijiazhuang, China
2Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
3Hebei Collaborative Innovation Center for Eco-Environment, Shijiazhuang, China

Tóm tắt

L-Carnosine is a natural biologically active dipeptide with critical physiological functions, such as antioxidant, antiglycation, and cytoplasmic buffering properties. Direct enzymatic synthesis is a promising way for L-carnosine production. In this study, a new aminopeptidase (gene_236976) with synthetic activity toward L-carnosine was identified by a metagenome mining approach from deep-sea sediment and functionally expressed in Escherichia coli. The enzyme shared a low identity of 14.3% with reported L-carnosine dipeptidase (SmPepD) from Serratia marcescens. β-Alanine methyl ester was proven to be the best substrate for the synthesis, and no ATP was needed for the enzymatic reaction. The enzyme activity was increased by structure-guided rational design. Only the mutant of G310 site gave positive results, and G310A mutant showed the best performance among the site-direct saturation mutagenesis, indicating that the additional CH3 group of mutant G310A was the main factor affecting the enzymatic activity. The engineered enzyme produced about 10 mM L-carnosine was produced from substrates of 50 mM β-alanine methyl ester and 50 mM L-histidine, under a tentatively optimized condition. This study enriched the enzyme resources for developing the microbial synthesis process of L-carnosine production.

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