Synthesis of N-Acetyl-trans-4-hydroxyproline Catalyzed by Acyltransferase

Abstract:

[Objective] This study aims to establish the biosynthesis process of N-acetyl-trans-4-hydroxyproline (N-ATHOP). [Method]Escherichia coli (E. coli) BL21 (DE3) was selected as the host to heterologously express the acyltransferase (MsAcT) derived from Mycobacterium smegmatis. A semi-rational design was employed to construct the mutant MsAcT(S11C) capable of catalyzing the synthesis of N-ATHOP. [Result] The catalytic reaction conditions of the mutant MsAcT(S11C) were optimized as pH 6.9, reaction temperature 49 ℃, initial trans-4-hydroxyproline (THOP) concentration of 315.5 mmol/L, and vinyl acetate volume fraction of 10%. Under the optimized conditions, the yield of N-ATHOP synthesized under the catalysis of MsAcT(S11C) reached 4.71 g/L, with a conversion rate of 10.95% after 12 h of reaction. [Conclusion] The mutant MsAcT(S11C) successfully achieved microbial enzymatic biosynthesis from THOP to N-ATHOP.