Premium
Customizing the Enantioselectivity of a Cyclohexanone Monooxygenase by a Strategy Combining “Size‐Probes” with in silico Study
Author(s) -
Hu Yujing,
Xu Jian,
Cen Yixin,
Li Danyang,
Zhang Yu,
Huang Meilan,
Lin Xianfu,
Wu Qi
Publication year - 2019
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201901200
Subject(s) - in silico , monooxygenase , cyclohexanone , combinatorial chemistry , chemistry , rational design , mutant , directed evolution , small molecule , protein engineering , molecule , stereochemistry , enzyme , nanotechnology , catalysis , biochemistry , organic chemistry , materials science , cytochrome p450 , gene
Enzymatic Baeyer‐Villiger oxidation provides a promising green route utilizing molecular oxygen as the oxidant to produce chiral lactones. Wild‐type (WT) CHMO Acineto leads to enantioselectivity up to 99 % ee ( S ) in the synthesis of substituted ϵ‐caprolactones. To reverse the inherent enantiopreference of CHMO Acineto toward an array of cyclohexanones with various chain length, we herein reshaped the binding pocket with a minimal number of mutations by a rational design strategy combining “size‐probes” with in silico study, which drastically reduces the screening effort. By probing the binding pocket of variants with different‐sized 4‐substituted cyclohexanones substrates, single, double and triple mutants were identified as the best mutants providing highly reversed enantioselectivity for these probing molecules, respectively. The successful demonstration of the strategy combining “size‐probes” with in silico study in the protein engineering of CHMO Acineto may provide a valuable guidance for facile engineering other BVMOs with customized enantiopreference for the same classified substrates with their substituents on the chiral or prochiral central atom.