Semi-rational protein engineering of a novel esterase from Bacillus aryabhattai (BaCE) for resolution of (R,S)-ethyl indoline-2-carboxylate to prepare (S)-indoline-2-carboxylic acid

Hongjun Zhang; Zeguang Cheng; Litian Wei; Xinjun Yu; Zhao Wang; Yinjun Zhang

doi:10.1016/j.bioorg.2022.105602

Semi-rational protein engineering of a novel esterase from Bacillus aryabhattai (BaCE) for resolution of (R,S)-ethyl indoline-2-carboxylate to prepare (S)-indoline-2-carboxylic acid

Bioorg Chem. 2022 Mar:120:105602. doi: 10.1016/j.bioorg.2022.105602. Epub 2022 Jan 10.

Authors

Hongjun Zhang¹, Zeguang Cheng¹, Litian Wei¹, Xinjun Yu², Zhao Wang¹, Yinjun Zhang³

Affiliations

¹ Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.
² Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China. Electronic address: xjyu@zjut.edu.cn.
³ Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China. Electronic address: zhangyj@zjut.edu.cn.

PMID: 35065466
DOI: 10.1016/j.bioorg.2022.105602

Abstract

A gene encoding an esterase from Bacillus aryabhattai (BaCE) was identified, synthesized and efficiently expressed in the Escherichia coli system. A semi-rational protein engineering was applied to further improve the enzyme's enantioselectivity. Under the guidance of the molecular docking result, a single mutant BaCE-L86Q and a double mutant BaCE-L86Q/G284E were obtained, with its E_max value 6.4 times and 13.9 times of the wild-type BaCE, respectively. The recombinant BaCEs were purified and characterized. The overwhelming E value demonstrated that BaCE-L86Q/G284E was a promising biocatalyst for the biological resolution to prepare (S)-indoline-2-carboxylic acid.

Keywords: (S)-indoline-2-carboxylic acid; Biocatalysis; Enantioselective; Esterase; Overexpression; Protein engineering.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacillus
Carboxylic Acids*
Escherichia coli / genetics
Escherichia coli / metabolism
Esterases* / metabolism
Indoles
Molecular Docking Simulation
Protein Engineering

Substances

Carboxylic Acids
Indoles
indoline-2-carboxylate
indoline
Esterases

Supplementary concepts

Bacillus aryabhattai