文献类型：期刊文献

英文题名：Engineering of halide methyltransferases for synthesis of SAE and its application in biosynthesis of ethyl vanillin

作者：Yang, Gui-Ying[1,2];Zheng, Gao-Wei[1,2];Zeng, Bu-Bing[3];Xu, Jian-He[1,2];Chen, Qi[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Sch Pharm, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Me, State Key Lab Bioreactor Engn,Optogenet & Synthet, Shanghai 200237, Peoples R China

年份：2023

卷号：550

外文期刊名：MOLECULAR CATALYSIS

收录：;EI(收录号：20233814764494);WOS:【SCI-EXPANDED(收录号:WOS:001082401200001)】；

基金：This work was financially supported by the National Key Research and Development Program of China (2021YFC2102300 and 2019YFA0905000) , the National Natural Science Foundation of China (31971380) .

语种：英文

外文关键词：Halide methyltransferase; Semi-rational design; Molecular dynamics simulation; Biocatalysis; Ethyl vanillin

摘要：Halide methyltransferase (HMT)-catalyzed synthesis of alkyl donors from S-adenosyl-L-homocysteine (SAH) and haloalkanes is a promising method for biocatalytic alkylation. However, the reported halide methyltransferases show limited catalytic capacity towards unnatural substrates such as ethyl iodide, which limits their application in biocatalytic ethylation. In this work, we performed structure-based directed evolution starting from the wildtype Aspergillus clavatus halide methyltransferase (AcHMT). The best variant, AcHMTM3, was identified with an activity of up to 421.5 mU mg-1 towards ethyl iodide, 38.7-fold higher than that of the wild type AcHMT. Subsequently, molecular dynamics simulations provide structural insights into how mutations improve the catalytic activity towards iodoethane. Finally, AcHMTM3 was utilized in one-pot cascade with the reported Omethyltransferase RnCOMT+Y200L to synthesize ethyl vanillin from 3,4-dihydroxybenzaldehyde (79% conversion). Our work demonstrated the potential of the engineered HMT in further biocatalytic alkylation.