Structure-guided protein design of fluoroacetate dehalogenase for kinetic resolution of rac-2-bromobutyric acid

Green Synthesis and Catalysis

卷:1

Xudong Qu¹  Jian-Bo Wang²  Yanbing Xu³  Zhanbing Cheng³  Zhenhua Tian³  Shu Wang³  Lu Yang⁴ 

[1] Corresponding authors.;

[2] Key Laboratory of Phytochemistry R&D of Hunan Province, and Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081, China;

[3] Shanghai R&D Center, Abiochem Co. Ltd., Shanghai 200241, China;

[4] State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;

关键词: Fluoroacetate dehalogenase;    Semi-rational design;    Enzyme engineering;    Kinetic resolution;    2-bromobutyric acid;   

DOI  :  

来源: DOAJ

【 摘 要 】

R-2-Bromobutyric acid is a very important intermediate for the synthesis of agrochemicals and pharmaceuticals. Bioresolution of rac-2-bromobutyric acid (rac-2-BBA) provides a promising process for R-2-bromobutyric acid (R-2-BBA) production. The fluoroacetate dehalogenase (FAcD) has been always studied in the defluorination process. We found that FAcD RPA1163 showed detectable activity but no enantioselectivity towards rac-2-BBA. The iterative saturation mutagenesis (ISM) of FAcD RPA1163 resulted in a mutant H155V/W156R/Y219M, which catalyzed the kinetic resolution of rac-2-BBA to produce R-2-BBA with enhanced activity and enantioselectivity (99.3% ee). The high preference for S-2-bromobutyric acid (S-2-BBA) is of synthetic value. Molecular docking analysis indicated that the H155V/W156R/Y219M mutation reduced steric hindrance and broadened the halide pocket. It is not only the steric hindrance but also the electrostatic environment that has an effect on the activity and enantioselectivity.

【 授权许可】

Unknown