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dc.contributor.author
Mansouri, Hamid R.
dc.contributor.author
Gracia Carmona, Oriol
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Jodlbauer, Julia
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Schweiger, Lorenz
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Fink, Michael J.
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Breslmayr, Erik
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Laurent, Christophe
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Feroz, Saima
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Goncalves, Leticia C. P.
dc.contributor.author
Rial, Daniela Veronica
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Mihovilovic, Marko D.
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Bommarius, Andreas S.
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Ludwig, Roland
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Oostenbrink, Chris
dc.contributor.author
Rudroff, Florian
dc.date.available
2023-11-15T16:05:56Z
dc.date.issued
2022-09
dc.identifier.citation
Mansouri, Hamid R.; Gracia Carmona, Oriol; Jodlbauer, Julia; Schweiger, Lorenz; Fink, Michael J.; et al.; Mutations Increasing Cofactor Affinity, Improve Stability and Activity of a Baeyer-Villiger Monooxygenase; American Chemical Society; ACS Catalysis; 12; 19; 9-2022; 11761-11766
dc.identifier.issn
2155-5435
dc.identifier.uri
http://hdl.handle.net/11336/218216
dc.description.abstract
The typically low thermodynamic and kinetic stability of enzymes is a bottleneck for their application in industrial synthesis. Baeyer-Villiger monooxygenases, which oxidize ketones to lactones using aerial oxygen, among other activities, suffer particularly from these instabilities. Previous efforts in protein engineering have increased thermodynamic stability but at the price of decreased activity. Here, we solved this trade-off by introducing mutations in a cyclohexanone monooxygenase from Acinetobacter sp., guided by a combination of rational and structure-guided consensus approaches. We developed variants with improved activity (1.5- to 2.5-fold) and increased thermodynamic (+5 °C Tm) and kinetic stability (8-fold). Our analysis revealed a crucial position in the cofactor binding domain, responsible for an 11-fold increase in affinity to the flavin cofactor, and explained using MD simulations. This gain in affinity was compatible with other mutations. While our study focused on a particular model enzyme, previous studies indicate that these findings are plausibly applicable to other BVMOs, and possibly to other flavin-dependent monooxygenases. These new design principles can inform the development of industrially robust, flavin-dependent biocatalysts for various oxidations.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
American Chemical Society
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by/2.5/ar/
dc.subject
CYCLOHEXANONE MONOOXYGENASE
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ENZYME STABILIZATION
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MUTAGENESIS
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OXIDATION
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PROTEIN ENGINEERING
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STRUCTURE-GUIDED CONSENSUS APPROACH
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Bioquímica y Biología Molecular
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Mutations Increasing Cofactor Affinity, Improve Stability and Activity of a Baeyer-Villiger Monooxygenase
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2023-11-14T14:30:26Z
dc.journal.volume
12
dc.journal.number
19
dc.journal.pagination
11761-11766
dc.journal.pais
Estados Unidos
dc.description.fil
Fil: Mansouri, Hamid R.. Vienna University of Technology; Austria
dc.description.fil
Fil: Gracia Carmona, Oriol. Universitat Fur Bodenkultur Wien; Austria
dc.description.fil
Fil: Jodlbauer, Julia. Vienna University of Technology; Austria
dc.description.fil
Fil: Schweiger, Lorenz. Universitat Fur Bodenkultur Wien; Austria
dc.description.fil
Fil: Fink, Michael J.. Vienna University of Technology; Austria
dc.description.fil
Fil: Breslmayr, Erik. Universitat Fur Bodenkultur Wien; Austria
dc.description.fil
Fil: Laurent, Christophe. Universitat Fur Bodenkultur Wien; Austria
dc.description.fil
Fil: Feroz, Saima. Vienna University of Technology; Austria
dc.description.fil
Fil: Goncalves, Leticia C. P.. Université Côte D'azur; Francia
dc.description.fil
Fil: Rial, Daniela Veronica. Universidad Nacional de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina
dc.description.fil
Fil: Mihovilovic, Marko D.. Vienna University of Technology; Austria
dc.description.fil
Fil: Bommarius, Andreas S.. Georgia Institute of Techology; Estados Unidos
dc.description.fil
Fil: Ludwig, Roland. Universitat Fur Bodenkultur Wien; Austria
dc.description.fil
Fil: Oostenbrink, Chris. Universitat Fur Bodenkultur Wien; Austria
dc.description.fil
Fil: Rudroff, Florian. Vienna University of Technology; Austria
dc.journal.title
ACS Catalysis
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1021/acscatal.2c03225
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