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.
Originalsprache | englisch |
---|---|
Seiten (von - bis) | 38-43 |
Seitenumfang | 6 |
Fachzeitschrift | ACS Chemical Biology |
Jahrgang | 11 |
Ausgabenummer | 1 |
DOIs | |
Publikationsstatus | Veröffentlicht - 15 Jan 2016 |
Extern publiziert | Ja |
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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
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TY - JOUR
T1 - Switching the Regioselectivity of a Cyclohexanone Monooxygenase toward (+)-trans-Dihydrocarvone by Rational Protein Design
AU - Balke, Kathleen
AU - Schmidt, Sandy
AU - Genz, Maika
AU - Bornscheuer, Uwe T.
PY - 2016/1/15
Y1 - 2016/1/15
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.
KW - Actinobacteria
KW - Arthrobacter
KW - Catalytic Domain
KW - Drug Design
KW - Models, Molecular
KW - Molecular Docking Simulation
KW - Monoterpenes
KW - Mutation
KW - Oxygenases
KW - Stereoisomerism
KW - Substrate Specificity
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1021/acschembio.5b00723
DO - 10.1021/acschembio.5b00723
M3 - Article
VL - 11
SP - 38
EP - 43
JO - ACS Chemical Biology
JF - ACS Chemical Biology
SN - 1554-8929
IS - 1
ER -