Laboratory Evolution of High-Redox Potential Laccases

12 páginas, 5 figuras, 2 tablas -- PAGS nros. 1030-1041

Detalles Bibliográficos
Autores: Maté, Diana M., García-Burgos, Carlos, García-Ruiz, Eva, Ballesteros Olmo, Antonio, Camarero, Susana, Alcalde Galeote, Miguel
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2010
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/46385
Acceso en línea:http://hdl.handle.net/10261/46385
Access Level:acceso abierto
Palabra clave:Lacasse
Chemistry
Directed Molecular Evolutions
Enzymology
Genetics
Metabolism
Oxidation Reduction Reaction
pH
Protein Stability
Protein Tertiary Structure
Saccharomyces Cerevisae
Site directed mutagenesis
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spelling Laboratory Evolution of High-Redox Potential LaccasesMaté, Diana M.García-Burgos, CarlosGarcía-Ruiz, EvaBallesteros Olmo, AntonioCamarero, SusanaAlcalde Galeote, MiguelLacasseChemistryDirected Molecular EvolutionsEnzymologyGeneticsMetabolismOxidation Reduction ReactionpHProtein StabilityProtein Tertiary StructureSaccharomyces CerevisaeSite directed mutagenesis12 páginas, 5 figuras, 2 tablas -- PAGS nros. 1030-1041Thermostable laccases with a high-redox potential have been engineered through a strategy that combines directed evolution with rational approaches. The original laccase signal sequence was replaced by the α-factor prepro-leader, and the corresponding fusion gene was targeted for joint laboratory evolution with the aim of improving kinetics and secretion by Saccharomyces cerevisiae, while retaining high thermostability. After eight rounds of molecular evolution, the total laccase activity was enhanced 34,000-fold culminating in the OB-1 mutant as the last variant of the evolution process, a highly active and stable enzyme in terms of temperature, pH range, and organic cosolvents. Mutations in the hydrophobic core of the evolved α-factor prepro-leader enhanced functional expression, whereas some mutations in the mature protein improved its catalytic capacities by altering the interactions with the surrounding residuesEU Projects (NMP4-SL-2009-229255, NMP2-CT-2006-026456, COST Action CM0701) and National projects (CTQ2005-08925-CO2-02, BIO2010-19697 and CCG08-CSIC/PPQ-3706). NeuronBiopharma for financial support through Research Contracts 020401070029 (Profit Program) and 020401070004 (Idea Program).Peer reviewedElsevierEuropean CommissionNeuron BiopharmaMinisterio de Economía y Competitividad (España)201220122010info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/46385reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/EC/FP7/229255http://dx.doi.org/10.1016/j.chembiol.2010.07.010info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/463852026-05-22T06:33:51Z
dc.title.none.fl_str_mv Laboratory Evolution of High-Redox Potential Laccases
title Laboratory Evolution of High-Redox Potential Laccases
spellingShingle Laboratory Evolution of High-Redox Potential Laccases
Maté, Diana M.
Lacasse
Chemistry
Directed Molecular Evolutions
Enzymology
Genetics
Metabolism
Oxidation Reduction Reaction
pH
Protein Stability
Protein Tertiary Structure
Saccharomyces Cerevisae
Site directed mutagenesis
title_short Laboratory Evolution of High-Redox Potential Laccases
title_full Laboratory Evolution of High-Redox Potential Laccases
title_fullStr Laboratory Evolution of High-Redox Potential Laccases
title_full_unstemmed Laboratory Evolution of High-Redox Potential Laccases
title_sort Laboratory Evolution of High-Redox Potential Laccases
dc.creator.none.fl_str_mv Maté, Diana M.
García-Burgos, Carlos
García-Ruiz, Eva
Ballesteros Olmo, Antonio
Camarero, Susana
Alcalde Galeote, Miguel
author Maté, Diana M.
author_facet Maté, Diana M.
García-Burgos, Carlos
García-Ruiz, Eva
Ballesteros Olmo, Antonio
Camarero, Susana
Alcalde Galeote, Miguel
author_role author
author2 García-Burgos, Carlos
García-Ruiz, Eva
Ballesteros Olmo, Antonio
Camarero, Susana
Alcalde Galeote, Miguel
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv European Commission
Neuron Biopharma
Ministerio de Economía y Competitividad (España)
dc.subject.none.fl_str_mv Lacasse
Chemistry
Directed Molecular Evolutions
Enzymology
Genetics
Metabolism
Oxidation Reduction Reaction
pH
Protein Stability
Protein Tertiary Structure
Saccharomyces Cerevisae
Site directed mutagenesis
topic Lacasse
Chemistry
Directed Molecular Evolutions
Enzymology
Genetics
Metabolism
Oxidation Reduction Reaction
pH
Protein Stability
Protein Tertiary Structure
Saccharomyces Cerevisae
Site directed mutagenesis
description 12 páginas, 5 figuras, 2 tablas -- PAGS nros. 1030-1041
publishDate 2010
dc.date.none.fl_str_mv 2010
2012
2012
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/46385
url http://hdl.handle.net/10261/46385
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/EC/FP7/229255
http://dx.doi.org/10.1016/j.chembiol.2010.07.010
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812455

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