Computational-Aided Engineering of a Selective Unspecific Peroxygenase toward Enantiodivergent β-Ionone Hydroxylation
dc.contributor.author
dc.date.accessioned
2023-11-03T13:08:09Z
dc.date.available
2023-11-03T13:08:09Z
dc.date.issued
2023-06-21
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dc.description.abstract
Unspecific peroxygenases (UPOs) perform oxyfunctionalizations for a wide range of substrates utilizing H2O2 without the need for further reductive equivalents or electron transfer chains. Tailoring these promising enzymes toward industrial application was intensely pursued in the last decade with engineering campaigns addressing the heterologous expression, activity, stability, and improvements in chemo- and regioselectivity. One hitherto missing integral part was the targeted engineering of enantioselectivity for specific substrates with poor starting enantioselectivity. In this work, we present the engineering of the short-type MthUPO toward the enantiodivergent hydroxylation of the terpene model substrate, β-ionone. Guided by computational modeling, we designed a small smart library and screened it with a GC–MS setup. After two rounds of iterative protein evolution, the activity increased up to 17-fold and reached a regioselectivity of up to 99.6% for the 4-hydroxy-β-ionone. Enantiodivergent variants were identified with enantiomeric ratios of 96.6:3.4 (R) and 0.3:99.7 (S), respectively
dc.description.sponsorship
J.M. thanks the Friedrich-Naumann-Stiftung für die Freiheit for a PhD scholarship. D.H. thanks the Friedrich-Ebert Stiftung for a PhD scholarship. M.J.W. and J.M. thank the Bundesministerium für Bildung und Forschung (Maßgeschneiderte Inhaltsstoffe 2, 031B0834A) and M.J.W., J.M., and D.H. thank the German Research Foundation (DFG, project ID 43649874, TP A05, RTG 2670) for generous funding. M.G.B. thanks the Spanish MICINN (Ministerio de Ciencia e Innovación) for PID2019-111300GA-I00 project and the Ramón y Cajal program via the RYC 2020-028628-I fellowship. J.S. thanks the Spanish MIU (Ministerio de Universidades) for a predoctoral FPU fellowship FPU18/02380
dc.format.mimetype
application/pdf
dc.language.iso
eng
dc.publisher
American Chemical Society (ACS)
dc.relation
PID2019-111300GA-I00
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Reproducció digital del document publicat a: https://doi.org/10.1021/acscatal.3c00702
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ACS Catalysis, 2023, vol. 13, núm. 13, p. 8963-8972
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Articles publicats (D-Q)
dc.rights
Attribution-NonCommercial-NoDerivatives 4.0 International
dc.rights.uri
dc.subject
dc.title
Computational-Aided Engineering of a Selective Unspecific Peroxygenase toward Enantiodivergent β-Ionone Hydroxylation
dc.type
info:eu-repo/semantics/article
dc.rights.accessRights
info:eu-repo/semantics/openAccess
dc.relation.projectID
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-111300GA-I00/ES/CARACTERIZACION MULTIESCALAR DE INTERMEDIOS REACTIVOS PARA EL DESCUBRIMIENTO Y DISEÑO DE NUEVAS ACTIVIDADES BIOCATALITICAS/
dc.type.version
info:eu-repo/semantics/publishedVersion
dc.identifier.doi
dc.identifier.idgrec
037198
dc.contributor.funder
dc.type.peerreviewed
peer-reviewed
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dc.identifier.eissn
2155-5435