Switching the Regioselectivity of a Cyclohexanone Monooxygenase toward (+)-trans-Dihydrocarvone by Rational Protein Design

Kathleen Balke, Sandy Schmidt, Maika Genz, Uwe T. Bornscheuer

Publikation: Beitrag in einer Fachzeitschrift › Artikel › Forschung › Begutachtung

Abstract

The regioselectivity of the Baeyer-Villiger monooxygenase-catalyzed oxidation is governed mostly by electronic effects leading to the migration of the higher substituted residue. However, in some cases, substrate binding occurs in a way that the less substituted residue lies in an antiperiplanar orientation to the peroxy bond in the Criegee intermediate yielding in the formation of the "abnormal" lactone product. We are the first to demonstrate a complete switch in the regioselectivity of the BVMO from Arthrobacter sp. (CHMOArthro) as exemplified for (+)-trans-dihydrocarvone by redesigning the active site of the enzyme. In the designed triple mutant, the substrate binds in an inverted orientation leading to a ratio of 99:1 in favor of the normal lactone instead of exclusive formation of the abnormal lactone in case of the wild type enzyme. In order to validate our computational study, the beneficial mutations were successfully transferred to the CHMO from Acinetobacter sp. (CHMOAcineto), again yielding in a complete switch of regioselectivity.

Originalspracheenglisch
Seiten (von - bis)38-43
Seitenumfang6
FachzeitschriftACS Chemical Biology
Jahrgang11
Ausgabenummer1
DOIs
PublikationsstatusVeröffentlicht - 15 Jan 2016
Extern publiziertJa

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Regioselectivity
Lactones
Switches
Arthrobacter
Proteins
Acinetobacter
Substrates
Enzymes
Mixed Function Oxygenases
Catalytic Domain
Oxidation
Mutation
cyclohexanone oxygenase
dihydrocarvone

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    Switching the Regioselectivity of a Cyclohexanone Monooxygenase toward (+)-trans-Dihydrocarvone by Rational Protein Design. / Balke, Kathleen; Schmidt, Sandy; Genz, Maika; Bornscheuer, Uwe T.

    in: ACS Chemical Biology, Jahrgang 11, Nr. 1, 15.01.2016, S. 38-43.

    Publikation: Beitrag in einer Fachzeitschrift › Artikel › Forschung › Begutachtung

    Balke, K, Schmidt, S, Genz, M & Bornscheuer, UT 2016, 'Switching the Regioselectivity of a Cyclohexanone Monooxygenase toward (+)-trans-Dihydrocarvone by Rational Protein Design' ACS Chemical Biology, Jg. 11, Nr. 1, S. 38-43. https://doi.org/10.1021/acschembio.5b00723
    Balke K, Schmidt S, Genz M, Bornscheuer UT. Switching the Regioselectivity of a Cyclohexanone Monooxygenase toward (+)-trans-Dihydrocarvone by Rational Protein Design. ACS Chemical Biology. 2016 Jan 15;11(1):38-43. https://doi.org/10.1021/acschembio.5b00723
    Balke, Kathleen ; Schmidt, Sandy ; Genz, Maika ; Bornscheuer, Uwe T. / Switching the Regioselectivity of a Cyclohexanone Monooxygenase toward (+)-trans-Dihydrocarvone by Rational Protein Design. in: ACS Chemical Biology. 2016 ; Jahrgang 11, Nr. 1. S. 38-43.
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    N2 - The regioselectivity of the Baeyer-Villiger monooxygenase-catalyzed oxidation is governed mostly by electronic effects leading to the migration of the higher substituted residue. However, in some cases, substrate binding occurs in a way that the less substituted residue lies in an antiperiplanar orientation to the peroxy bond in the Criegee intermediate yielding in the formation of the "abnormal" lactone product. We are the first to demonstrate a complete switch in the regioselectivity of the BVMO from Arthrobacter sp. (CHMOArthro) as exemplified for (+)-trans-dihydrocarvone by redesigning the active site of the enzyme. In the designed triple mutant, the substrate binds in an inverted orientation leading to a ratio of 99:1 in favor of the normal lactone instead of exclusive formation of the abnormal lactone in case of the wild type enzyme. In order to validate our computational study, the beneficial mutations were successfully transferred to the CHMO from Acinetobacter sp. (CHMOAcineto), again yielding in a complete switch of regioselectivity.

    AB - The regioselectivity of the Baeyer-Villiger monooxygenase-catalyzed oxidation is governed mostly by electronic effects leading to the migration of the higher substituted residue. However, in some cases, substrate binding occurs in a way that the less substituted residue lies in an antiperiplanar orientation to the peroxy bond in the Criegee intermediate yielding in the formation of the "abnormal" lactone product. We are the first to demonstrate a complete switch in the regioselectivity of the BVMO from Arthrobacter sp. (CHMOArthro) as exemplified for (+)-trans-dihydrocarvone by redesigning the active site of the enzyme. In the designed triple mutant, the substrate binds in an inverted orientation leading to a ratio of 99:1 in favor of the normal lactone instead of exclusive formation of the abnormal lactone in case of the wild type enzyme. In order to validate our computational study, the beneficial mutations were successfully transferred to the CHMO from Acinetobacter sp. (CHMOAcineto), again yielding in a complete switch of regioselectivity.

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    KW - Models, Molecular

    KW - Molecular Docking Simulation

    KW - Monoterpenes

    KW - Mutation

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    KW - Stereoisomerism

    KW - Substrate Specificity

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