文献类型：期刊文献

英文题名：Energy and conformation determine the enantioselectivity of enzyme

作者：Gao, Bei[1];Jiang, Shuiqin[1];Wang, Liuzhu[1];Zhang, Lujia[2,3];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Normal Univ, Sch Chem & Mol Engn, Shanghai 200062, Peoples R China;[3]NYU Shanghai, NYU ECNU Ctr Computat Chem, Shanghai 200062, Peoples R China

年份：2018

卷号：129

起止页码：106

外文期刊名：BIOCHEMICAL ENGINEERING JOURNAL

收录：;EI(收录号：20174604397159);WOS:【SCI-EXPANDED(收录号:WOS:000418987700013)】；

基金：This work was funded by the National Natural Foundation of China (No. 31772007, and No. 31571786), Natural Science Foundation of Shanghai (No. 16ZR1449500), Special Program for Applied Research on Super Computation of the NSFC-Guangdong Joint Fund (the second phase) under Grant No. U1501501, and Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Chiral selectivity; Free energy; Structural conformation; Rational design; Lipase

摘要：Enantioselectivity of biocatalysts is a crucial property utilized for the synthesis of stereo-specific products. Owing to its complex mechanism, the development of precise and simple way to understand and alter enantioselectivity has been a significant pursuit and challenge for decades. In this study, an accurate calculation by MM/PBSA method was applied to investigate the binding free energy of R- and S-type substrates with enzyme. Meanwhile, a simple distance standard was developed to evaluates the binding conformation. Furthermore, we proposed that the binding free energy and binding conformation had significant effect on enzyme enantioselectivity simultaneously, and played pivotal role in K-m and k(cat), respectively. A well-studied, important lipase LipK107, and both positive and negative mutants were explored to validate our finding. Compared with WT, the mutations indeed resulted in the alteration of both binding energy and conformation, which collectively led to the changes of enzyme enantioselectivity. Our work indicated that rational enzyme engineer should attach great importance on both the binding free energy and the binding conformation of substrate and enzyme. Besides, since the binding energy and conformation could be calculated precisely, a promising large-scale in silico design strategy could be applied to gain various enzymes with outstanding characteristics. (C) 2017 Elsevier B.V. All rights reserved.