文献类型：期刊文献

英文题名：Carving the Active Site of CYP153A7 Monooxygenase for Improving Terminal Hydroxylation of Medium-Chain Fatty Acids

作者：Dong, Ya-Li[1,2,3];Chong, Gang-Gang[1,2,3];Li, Chun-Xiu[1,2,3];Chen, Qi[1,2,3];Pan, Jiang[1,2,3];Li, Ai-Tao[4];Xu, Jian-He[1,2,3]

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, Frontiers Sci Ctr Materiobiol & Dynam Chem, Shanghai 200237, Peoples R China;[4]Hubei Univ, Coll Life Sci, Hubei Collaborat Innovat Ctr Green Transformat, Bioresources Hubei Key Lab Ind Biotechnol, Wuhan 430062, Peoples R China

年份：2022

卷号：23

期号：9

外文期刊名：CHEMBIOCHEM

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000772240400001)】；

基金：This work was financially sponsored by the National Key R & D Program of China (2019YFA09005000), the National Natural Science Foundation of China, (21536004, 21776085, 21871085, 31971380 and 31971380), and the Fundamental Research Funds for the Central Universities (222201714026).

语种：英文

外文关键词：biocatalysis; CYP153A subfamily monooxygenases; fatty acids; protein engineering; terminal hydroxylation

摘要：The P450-mediated terminal hydroxylation of non-activated C-H bonds is a chemically challenging reaction. CYP153A7 monooxygenase, discovered in Sphingomonas sp. HXN200, belongs to the CYP153A subfamily and shows a pronounced terminal selectivity. Herein, we report the significantly improved terminal hydroxylation activity of CYP153A7 by redesign of the substrate binding pocket based on molecular docking of CYP153A7-C-8:0 and sequence alignments. Some of the resultant single mutants were advantageous over the wild-type enzyme with higher reaction rates, achieving a complete conversion of n-octanoic acid (C-8:0,C- 1 mM) in a shorter time period. Especially, a single-mutation variant, D258E, showed 3.8-fold higher catalytic efficiency than the wild type toward the terminal hydroxylation of medium-chain fatty acid C-8:0 to the high value-added product 8-hydroxyoctanoic acid.