文献类型：期刊文献

英文题名：Rational design to improve activity of the Est3563 esterase from Acinetobacter sp. LMB-5

作者：Ren, Li-Quan[2];Chang, Tian-Tian[2];Ren, Da-Peng[3];Zhou, Ying[2];Ye, Bang-Ce[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biosyst & Microanal, Meilong RD 130, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Sch Bioengn, Dept Food Sci & Technol, Meilong RD 130, Shanghai 200237, Peoples R China;[3]Klenzan Chem Co Ltd, Shanghai, Peoples R China

年份：2019

卷号：131

外文期刊名：ENZYME AND MICROBIAL TECHNOLOGY

收录：;EI(收录号：20193707422522);WOS:【SCI-EXPANDED(收录号:WOS:000496607400019)】；

基金：This research was supported by the Shanghai Municipal Science and Technology Commission Project (18JC1410802) and Fundamental Research Funds for the Central Universities (222201714051).

语种：英文

外文关键词：Esterase; Catalytic pocket; Rational design; 3D printing; Simulation mutagenesis

摘要：Acinetobacter sp. strain LMB-5 can produce a kind of esterase degrading phthalate esters. However, low activity of Est3563 esterase limited its large-scale application. In this study, computer-aided simulation mutagenesis was used to improve the esterase activity with a tightened screening library and enlarged success rate. Two positive mutants, P218R and A242R, were obtained with 2.5 and 2.1 folds higher than the WT Est3563 esterase, with 11.96 +/- 0.45 U.mg(-1) and 9.90 +/- 0.52 U.mg(-1), respectively. With the help of bioinformatics analysis and three-dimensional printing technology, it was found that the mutations could increase the 240-280 residues swing distance and make them deviate from the catalytic pocket. The instability and deviation of these residues on the lid-like structure of the esterase could deteriorate the seal of the binding pocket and expose the active site. Thus, the catalytic efficiency of the mutants became higher. This result demonstrates that the instability and deviation of the lid-like structure could expand the binding pocket of the esterase and enhance the esterase activity.