Premium
Inverting the Enantiopreference of Nitrilase‐Catalyzed Desymmetric Hydrolysis of Prochiral Dinitriles by Reshaping the Binding Pocket with a Mirror‐Image Strategy
Author(s) -
Yu Shanshan,
Li Jinlong,
Yao Peiyuan,
Feng Jinhui,
Cui Yunfeng,
Li Jianjiong,
Liu Xiangtao,
Wu Qiaqing,
Lin Jianping,
Zhu Dunming
Publication year - 2021
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202012243
Subject(s) - nitrilase , saturated mutagenesis , chemistry , stereochemistry , stereoselectivity , hydrolysis , enzyme , thienamycin , mutant , substrate (aquarium) , catalysis , binding pocket , combinatorial chemistry , biochemistry , oceanography , gene , geology , antibiotics
Abstract A mirror‐image strategy, that is, symmetry analysis of the substrate‐binding pocket, was applied to identify two key amino acid residues W170 and V198 that possibly modulate the enantiopreference of a nitrilase from Synechocystis sp. PCC6803 towards 3‐isobutyl glutaronitrile ( 1 a ). Exchange of these two residues resulted in the enantiopreference inversion (S, 90 % ee to R, 47 % ee). By further reshaping the substrate‐binding pocket via routine site‐saturation and combinatorial mutagenesis, variant E8 with higher activity and stereoselectivity (99 % ee, R) was obtained. The mutant enzyme was applied in the preparation of optically pure (R)‐3‐isobutyl‐4‐cyanobutanoic acid ((R)‐ 2 a ) and showed similar stereopreference inversion towards a series of 3‐substituted glutaronitriles. This study may offer a general strategy to switch the stereopreference of other nitrilases and other enzymes toward the desymmetric reactions of prochiral substrates with two identical reactive functional groups.