Customizing the Enantioselectivity of a Cyclohexanone Monooxygenase by a Strategy Combining “Size-probes” with in silico Study

Yujing Hu, Jian Xu, Yixin Cen, Danyang Li, Yu Zhang, Meilan Huang, Xianfu Lin, Qi Wu

Research output: Contribution to journalArticle

Abstract

Enzymatic Baeyer‐Villiger oxidation provides a promising green route utilizing molecular oxygen as the oxidant to produce chiral lactones. Wild‐type (WT) CHMOAcineto leads to enantioselectivity up to 99 % ee (S) in the synthesis of substituted ϵ‐caprolactones. To reverse the inherent enantiopreference of CHMOAcineto 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 CHMOAcineto 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.
Original languageEnglish
JournalCHEMCATCHEM
Early online date25 Aug 2019
DOIs
Publication statusEarly online date - 25 Aug 2019

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Cyclohexanones
Enantioselectivity
engineering
Molecular oxygen
probes
Lactones
Substrates
mutations
Chain length
Oxidants
Screening
Demonstrations
screening
routes
proteins
Proteins
Atoms
Oxidation
oxidation
Molecules

Cite this

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title = "Customizing the Enantioselectivity of a Cyclohexanone Monooxygenase by a Strategy Combining “Size-probes” with in silico Study",
abstract = "Enzymatic Baeyer‐Villiger oxidation provides a promising green route utilizing molecular oxygen as the oxidant to produce chiral lactones. Wild‐type (WT) CHMOAcineto leads to enantioselectivity up to 99 {\%} ee (S) in the synthesis of substituted ϵ‐caprolactones. To reverse the inherent enantiopreference of CHMOAcineto 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 CHMOAcineto 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.",
author = "Yujing Hu and Jian Xu and Yixin Cen and Danyang Li and Yu Zhang and Meilan Huang and Xianfu Lin and Qi Wu",
year = "2019",
month = "8",
day = "25",
doi = "10.1002/cctc.201901200",
language = "English",
journal = "CHEMCATCHEM",
issn = "1867-3880",
publisher = "Wiley - VCH Verlag GmbH & CO. KGaA",

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Customizing the Enantioselectivity of a Cyclohexanone Monooxygenase by a Strategy Combining “Size-probes” with in silico Study. / Hu, Yujing; Xu, Jian; Cen, Yixin; Li, Danyang ; Zhang, Yu; Huang, Meilan; Lin, Xianfu; Wu, Qi.

In: CHEMCATCHEM, 25.08.2019.

Research output: Contribution to journalArticle

TY - JOUR

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AU - Zhang, Yu

AU - Huang, Meilan

AU - Lin, Xianfu

AU - Wu, Qi

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