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

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

Research output: Contribution to journalArticleResearchpeer-review

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.

Original languageEnglish
Pages (from-to)38-43
Number of pages6
JournalACS Chemical Biology
Volume11
Issue number1
DOIs
Publication statusPublished - 15 Jan 2016
Externally publishedYes

Fingerprint

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

Keywords

  • Actinobacteria
  • Arthrobacter
  • Catalytic Domain
  • Drug Design
  • Models, Molecular
  • Molecular Docking Simulation
  • Monoterpenes
  • Mutation
  • Oxygenases
  • Stereoisomerism
  • Substrate Specificity
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

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, Vol. 11, No. 1, 15.01.2016, p. 38-43.

Research output: Contribution to journalArticleResearchpeer-review

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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 - Drug Design

KW - Models, Molecular

KW - Molecular Docking Simulation

KW - Monoterpenes

KW - Mutation

KW - Oxygenases

KW - Stereoisomerism

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