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Re-designing the substrate binding pocket of laccase for enhanced oxidation of sinapic acid
dc.contributor.author | Pardo, Isabel |
dc.contributor.author | Santiago, Gerard |
dc.contributor.author | Gentili, Patrizia |
dc.contributor.author | Lucas, Fátima |
dc.contributor.author | Monza, Emanuele |
dc.contributor.author | Medrano, Francisco Javier |
dc.contributor.author | Galli, Carlo |
dc.contributor.author | Martínez, Angel T. |
dc.contributor.author | Guallar, Víctor |
dc.contributor.author | Camarero, Susana |
dc.contributor.other | Barcelona Supercomputing Center |
dc.date.accessioned | 2016-03-11T10:47:07Z |
dc.date.available | 2016-03-11T10:47:07Z |
dc.date.issued | 2015-12-29 |
dc.identifier.citation | Pardo, 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.issn | 2044-4753 |
dc.identifier.uri | http://hdl.handle.net/2117/84199 |
dc.description.abstract | Iterative 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.sponsorship | This 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.extent | 11 p. |
dc.language.iso | eng |
dc.publisher | Royal Society of Chemistry |
dc.rights | Attribution-NonCommercial-NoDerivs 4.0 Spain |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/es/ |
dc.subject | Àrees temàtiques de la UPC::Enginyeria mecànica::Impacte ambiental |
dc.subject.lcsh | Oxidation--Measurement |
dc.subject.lcsh | Molecular dynamics |
dc.subject.other | Enhanced oxidation |
dc.subject.other | Sinapic acid |
dc.subject.other | Iterative saturation mutagenesis |
dc.title | Re-designing the substrate binding pocket of laccase for enhanced oxidation of sinapic acid |
dc.type | Article |
dc.subject.lemac | Dinàmica molecular |
dc.identifier.doi | 10.1039/C5CY01725D |
dc.description.peerreviewed | Peer Reviewed |
dc.relation.publisherversion | http://pubs.rsc.org/en/content/articlelanding/2015/cy/c5cy01725d#!divAbstract |
dc.rights.access | Open Access |
dc.description.version | Postprint (published version) |
dc.relation.projectid | info:eu-repo/grantAgreement/EC/FP7/613549/EU/Optimized oxidoreductases for medium and large scale industrial biotransformations/INDOX |
dc.relation.projectid | info:eu-repo/grantAgreement/MINECO//BIO2014-56388-R/ES/NUEVAS ENZIMAS OXIDATIVAS PARA UNA INDUSTRIA SOSTENIBLE/ |
dc.relation.projectid | info:eu-repo/grantAgreement/MINECO//CTQ2013-48287-R/ES/DISENYO COMPUTACIONAL RACIONAL DE OXIDOREDUCTASAS PARA APLICACIONES INDUSTRIALES Y TECNOLOGICAS/ |
local.citation.publicationName | Catalysis Science & Technology |
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