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

作　　者：Shu Wang Zhanbing Cheng Yanbing Xu Lu Yang Jian-Bo Wang Zhenhua Tian Xudong Qu 

机构地区：[1]Shanghai R&D Center,Abiochem Co.Ltd.,Shanghai 200241,China [2]State Key Laboratory of Microbial Metabolism,School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China [3]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

出　　处：《Green Synthesis and Catalysis》2020年第1期60-65,共6页绿色合成与催化（英文）

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.

关 键 词：Fluoroacetate dehalogenase Semi-rational design Enzyme engineering Kinetic resolution 2-bromobutyric acid 

分 类 号：O62[理学—有机化学]