文献类型：期刊文献

英文题名：Engineering of an oleate hydratase for efficient C10-Functionalization of oleic acid

作者：Sun, Qi-Fan[1];Zheng, Yu-Cong[1];Chen, Qi[1,2];Xu, Jian-He[1,2];Pan, Jiang[1,2]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomafg, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Frontiers Sci Ctr Mat & Dynam Chem, Shanghai 200237, Peoples R China

年份：2021

卷号：537

起止页码：64

外文期刊名：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000609438800010)】；

基金：This work was financially supported by the National Key R&D Program of China (2019YFA09005000 & 2018YFC1706200), the National Natural Science Foundation of China (21536004, 21776085, 21871085 and 31971380), and the Fundamental Research Funds for the Central Universities (22221818014).

语种：英文

外文关键词：Biocatalysis; Oleate hydratase; Directed evolution; Oleic acid; Fatty acid functionalization

摘要：Oleate hydratase catalyzes the hydration of unsaturated fatty acids, giving access to C-10-functionalization of oleic acid. The resultant 10-hydroxystearic acid is a key material for the synthesis of many biomass derived value-added products. Herein, we report the engineering of an oleate hydratase from Paracoccus aminophilus (PaOH) with significantly improved catalytic efficiency (from 33 s(-1) mM(-1) to 119 s(-1) mM(-1)), as well as 3.4 times increased half-life at 30 degrees C. The structural mechanism regarding the impact of mutations on the improved catalytic activity and thermostability was elucidated with the aid of molecular dynamics simulation. The practical feasibility of the engineered PaOH variant F233L/F122L/T15 N was demonstrated through the pilot synthesis of 10-hydroxystearic acid and 10-oxostearic acid via an optimized multi-enzymatic cascade reaction, with space-time yields of 540 g L-1 day(-1) and 160 g L-1 day(-1), respectively. (C) 2020 Published by Elsevier Inc.