Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases

Ancestral sequence reconstruction and resurrection provides useful information for protein engineering, yet its alliance with directed evolution has been little explored. In this study, we have resurrected several ancestral nodes of fungal laccases dating back ∼500 to 250 million years. Unlike moder...

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Detalhes bibliográficos
Autores: Gómez-Fernández, Bernardo J., Risso, Valeria A., Rueda, Andres, Sánchez-Ruiz, José M., Alcalde Galeote, Miguel
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/221368
Acesso em linha:http://hdl.handle.net/10261/221368
Access Level:acceso abierto
Palavra-chave:Ancestral reconstruction
Directed evolution
Laccase
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spelling Ancestral Resurrection and Directed Evolution of Fungal Mesozoic LaccasesGómez-Fernández, Bernardo J.Risso, Valeria A.Rueda, AndresSánchez-Ruiz, José M.Alcalde Galeote, MiguelAncestral reconstructionDirected evolutionLaccaseAncestral sequence reconstruction and resurrection provides useful information for protein engineering, yet its alliance with directed evolution has been little explored. In this study, we have resurrected several ancestral nodes of fungal laccases dating back ∼500 to 250 million years. Unlike modern laccases, the resurrected Mesozoic laccases were readily secreted by yeast, with similar kinetic parameters, a broader stability, and distinct pH activity profiles. The resurrected Agaricomycetes laccase carried 136 ancestral mutations, a molecular testimony to its origin, and it was subjected to directed evolution in order to improve the rate of 1,3-cyclopentanedione oxidation, a β–diketone initiator commonly used in vinyl polymerization reactions.This study is based upon work funded by the Spanish Government projects BIO2013-43407-R-DEWRY and BIO2016-79106-R-Lignolution and by the Bio Based Industries Joint Undertaking under the European Union’s Horizon 2020 Research and Innovation programme (grant agreement no. 886567). B.J.G.-F. was supported by FPI national fellowship BES-2014-068887.Peer reviewedAmerican Society for MicrobiologyMinisterio de Economía y Competitividad (España)European CommissionAlcalde Galeote, Miguel [0000-0001-6780-7616]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202020202020info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/221368reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2013-43407-R-DEWRYinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-79106-R-Lignolutioninfo:eu-repo/grantAgreement/EC/H2020/886567https://doi.org/10.1128/AEM.00778-20Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2213682026-05-22T06:33:51Z
dc.title.none.fl_str_mv Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
title Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
spellingShingle Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
Gómez-Fernández, Bernardo J.
Ancestral reconstruction
Directed evolution
Laccase
title_short Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
title_full Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
title_fullStr Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
title_full_unstemmed Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
title_sort Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases
dc.creator.none.fl_str_mv Gómez-Fernández, Bernardo J.
Risso, Valeria A.
Rueda, Andres
Sánchez-Ruiz, José M.
Alcalde Galeote, Miguel
author Gómez-Fernández, Bernardo J.
author_facet Gómez-Fernández, Bernardo J.
Risso, Valeria A.
Rueda, Andres
Sánchez-Ruiz, José M.
Alcalde Galeote, Miguel
author_role author
author2 Risso, Valeria A.
Rueda, Andres
Sánchez-Ruiz, José M.
Alcalde Galeote, Miguel
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
European Commission
Alcalde Galeote, Miguel [0000-0001-6780-7616]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Ancestral reconstruction
Directed evolution
Laccase
topic Ancestral reconstruction
Directed evolution
Laccase
description Ancestral sequence reconstruction and resurrection provides useful information for protein engineering, yet its alliance with directed evolution has been little explored. In this study, we have resurrected several ancestral nodes of fungal laccases dating back ∼500 to 250 million years. Unlike modern laccases, the resurrected Mesozoic laccases were readily secreted by yeast, with similar kinetic parameters, a broader stability, and distinct pH activity profiles. The resurrected Agaricomycetes laccase carried 136 ancestral mutations, a molecular testimony to its origin, and it was subjected to directed evolution in order to improve the rate of 1,3-cyclopentanedione oxidation, a β–diketone initiator commonly used in vinyl polymerization reactions.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/221368
url http://hdl.handle.net/10261/221368
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2013-43407-R-DEWRY
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-79106-R-Lignolution
info:eu-repo/grantAgreement/EC/H2020/886567
https://doi.org/10.1128/AEM.00778-20
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
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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