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Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants

Saturation mutagenesis was performed over six residues delimiting the substrate binding pocket of a fungal laccase previously engineered in the lab. Mutant libraries were screened using sinapic acid as a model substrate, and those mutants presenting increased activity were selected for exploring the...

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Detalles Bibliográficos
Autores principales: Pardo, Isabel, Camarero, Susana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6332420/
https://www.ncbi.nlm.nih.gov/pubmed/26364626
http://dx.doi.org/10.3390/molecules200915929
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author Pardo, Isabel
Camarero, Susana
author_facet Pardo, Isabel
Camarero, Susana
author_sort Pardo, Isabel
collection PubMed
description Saturation mutagenesis was performed over six residues delimiting the substrate binding pocket of a fungal laccase previously engineered in the lab. Mutant libraries were screened using sinapic acid as a model substrate, and those mutants presenting increased activity were selected for exploring the oxidation of lignin-derived phenols. The latter comprised a battery of phenolic compounds of interest due to their use as redox mediators or precursors of added-value products and their biological activity. The new laccase variants were investigated in a multi-screening assay and the structural determinants, at both the substrate and the protein level, for the oxidation of the different phenols are discussed. Laccase activity greatly varied only by changing one or two residues of the enzyme pocket. Our results suggest that once the redox potential threshold is surpassed, the contribution of the residues of the enzymatic pocket for substrate recognition and binding strongly influence the overall rate of the catalytic reaction.
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spelling pubmed-63324202019-01-24 Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants Pardo, Isabel Camarero, Susana Molecules Article Saturation mutagenesis was performed over six residues delimiting the substrate binding pocket of a fungal laccase previously engineered in the lab. Mutant libraries were screened using sinapic acid as a model substrate, and those mutants presenting increased activity were selected for exploring the oxidation of lignin-derived phenols. The latter comprised a battery of phenolic compounds of interest due to their use as redox mediators or precursors of added-value products and their biological activity. The new laccase variants were investigated in a multi-screening assay and the structural determinants, at both the substrate and the protein level, for the oxidation of the different phenols are discussed. Laccase activity greatly varied only by changing one or two residues of the enzyme pocket. Our results suggest that once the redox potential threshold is surpassed, the contribution of the residues of the enzymatic pocket for substrate recognition and binding strongly influence the overall rate of the catalytic reaction. MDPI 2015-09-02 /pmc/articles/PMC6332420/ /pubmed/26364626 http://dx.doi.org/10.3390/molecules200915929 Text en © 2015 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pardo, Isabel
Camarero, Susana
Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
title Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
title_full Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
title_fullStr Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
title_full_unstemmed Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
title_short Exploring the Oxidation of Lignin-Derived Phenols by a Library of Laccase Mutants
title_sort exploring the oxidation of lignin-derived phenols by a library of laccase mutants
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6332420/
https://www.ncbi.nlm.nih.gov/pubmed/26364626
http://dx.doi.org/10.3390/molecules200915929
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