文献类型：期刊文献

英文题名：Engineering P450LaMO stereospecificity and product selectivity for selective C-H oxidation of tetralin-like alkylbenzenes

作者：Li, Ren-Jie[1];Li, Aitao[2];Zhao, Jing[3];Chen, Qi[1];Li, Ning[4];Yu, Hui-Lei[1];Xu, Jian-He[1]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Hubei Univ, Hubei Collaborat Innovat Ctr Green Transformat Bi, Coll Life Sci, Hubei Key Lab Ind Biotechnol, Wuhan 430062, Hubei, Peoples R China;[3]Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China;[4]South China Univ Technol, Coll Light Ind & Food Sci, State Key Lab Pulp & Paper Engn, Guangzhou 510640, Guangdong, Peoples R China

年份：2018

卷号：8

期号：18

起止页码：4638

外文期刊名：CATALYSIS SCIENCE & TECHNOLOGY

收录：;EI(收录号：20183905863327);WOS:【SCI-EXPANDED(收录号:WOS:000448144600008)】；

基金：This work was financially supported by the National Natural Science Foundation of China (Nos. 21672063 & 21536004), the Fundamental Research Fund for the Central Universities (No. 222201514039), and the Science and Technology Commission of Shanghai Municipality (No. 15JC1400403). We thank Taiwo Dele-Osibanjo (Tianjin Institute of Industrial Biotechnology) for discussing the manuscript.

语种：英文

外文关键词：Ketones - Genetic engineering - Amino acids - Biochemical engineering - Proteins

摘要：The P450-mediated asymmetric hydroxylation of inert C-H bonds is a chemically challenging reaction. Self-sufficient P450(LaMO) from the CYP116B subfamily could catalyze the transformation of 1,2,3,4-tetrahydronaphthalene to (S)-tetralol, despite its poor enantioselectivity (er 66:34) and product selectivity (the ratio of alcohol and ketone, ak, 76:24). To improve the selectivity, phenylalanine scanning and further protein engineering were performed to reshape the active pocket of P450(LaMO), resulting in a mutant (T121V/Y385F/M391L) with not only improved (S)-enantioselectivity (er 98:2) but also excellent product selectivity (ak 99:1), in contrast to another mutant L97F/T121F/E282V/T283Y with complementary (R)-enantioselectivity (er 23:77). Moreover, the enantiopure (S)-alcohols formed by the P450(LaMO)-catalyzed oxidation of a series of alkylbenzenes are potentially important building blocks in the pharmaceutical industry. This Phe-based enantioselectivity engineering used for reshaping the active pocket of P450s could provide a guide to the protein evolution of other CYP116B members.