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dc.contributor.authorPardo, Isabel
dc.contributor.authorSantiago, Gerard
dc.contributor.authorGentili, Patrizia
dc.contributor.authorLucas, Fátima
dc.contributor.authorMonza, Emanuele
dc.contributor.authorMedrano, Francisco Javier
dc.contributor.authorGalli, Carlo
dc.contributor.authorMartínez, Angel T.
dc.contributor.authorGuallar, Víctor
dc.contributor.authorCamarero, Susana
dc.contributor.otherBarcelona Supercomputing Center
dc.date.accessioned2016-03-11T10:47:07Z
dc.date.available2016-03-11T10:47:07Z
dc.date.issued2015-12-29
dc.identifier.citationPardo, Isabel [et al.]. Re-designing the substrate binding pocket of laccase for enhanced oxidation of sinapic acid. "Catalysis Science & Technology", 29 Desembre 2015.
dc.identifier.issn2044-4753
dc.identifier.urihttp://hdl.handle.net/2117/84199
dc.description.abstractIterative saturation mutagenesis was performed over six residues delimiting the substrate binding pocket of a high redox potential chimeric laccase with the aim of enhancing its activity over sinapic acid, a ligninrelated phenol of industrial interest. In total, more than 15000 clones were screened and two selected variants, together with the parent-type laccase, were purified and characterized. The new variants presented shifted pH activity profiles and enhanced turnover rates on sinapic acid and its methyl ester, whereas the oxidation of related phenols was not significantly enhanced. Neither the enzyme's redox potential nor the mechanism of the reaction was affected. Quantum mechanics and molecular dynamics calculations were done to rationalize the effect of the selected mutations, revealing the critical role of the residues of the enzyme pocket to provide the precise binding of the substrate that enables an efficient electron transfer to the T1 copper. The results presented highlight the power of combining directed evolution and computational approaches to give novel solutions in enzyme engineering and to understand the mechanistic reasons behind them, offering new insights for further rational design towards specific targets.
dc.description.sponsorshipThis work was funded by INDOX (KBBE-2013-7-613549) European project and NOESIS (BIO2014-56388-R) and CTQ2013- 48287-R Spanish National Projects. I. P. and G. S. acknowledge the Spanish Research Council (CSIC) and MINECO for their respective predoctoral fellowships.
dc.format.extent11 p.
dc.language.isoeng
dc.publisherRoyal Society of Chemistry
dc.rightsAttribution-NonCommercial-NoDerivs 4.0 Spain
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/es/
dc.subjectÀrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambiental
dc.subject.lcshOxidation--Measurement
dc.subject.lcshMolecular dynamics
dc.subject.otherEnhanced oxidation
dc.subject.otherSinapic acid
dc.subject.otherIterative saturation mutagenesis
dc.titleRe-designing the substrate binding pocket of laccase for enhanced oxidation of sinapic acid
dc.typeArticle
dc.subject.lemacDinàmica molecular
dc.identifier.doi10.1039/C5CY01725D
dc.description.peerreviewedPeer Reviewed
dc.relation.publisherversionhttp://pubs.rsc.org/en/content/articlelanding/2015/cy/c5cy01725d#!divAbstract
dc.rights.accessOpen Access
dc.description.versionPostprint (published version)
dc.relation.projectidinfo:eu-repo/grantAgreement/EC/FP7/613549/EU/Optimized oxidoreductases for medium and large scale industrial biotransformations/INDOX
dc.relation.projectidinfo:eu-repo/grantAgreement/MINECO/1PE/BIO2014-56388-R
dc.relation.projectidinfo:eu-repo/grantAgreement/MINECO/1PE/CTQ2013-48287-R
upcommons.citation.publishedtrue
upcommons.citation.publicationNameCatalysis Science & Technology


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