Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase

9 p.-8 fig.-2 tab.

Detalhes bibliográficos
Autores: Carro, Juan, González-Benjumea, Alejandro, Fernández-Fueyo, Elena, Aranda, Carmen, Guallar, Victor, Gutiérrez Suárez, Ana, Martínez, Ángel T.
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/184640
Acesso em linha:http://hdl.handle.net/10261/184640
Access Level:acceso abierto
Palavra-chave:Unspecific peroxygenase
Marasmius rotula
Unsaturated fatty acids
Epoxidation
Hydroxylation
Computational simulations
PELE software
Gas chromatography−mass spectrometry
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oai_identifier_str oai:digital.csic.es:10261/184640
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
title Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
spellingShingle Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
Carro, Juan
Unspecific peroxygenase
Marasmius rotula
Unsaturated fatty acids
Epoxidation
Hydroxylation
Computational simulations
PELE software
Gas chromatography−mass spectrometry
title_short Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
title_full Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
title_fullStr Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
title_full_unstemmed Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
title_sort Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenase
dc.creator.none.fl_str_mv Carro, Juan
González-Benjumea, Alejandro
Fernández-Fueyo, Elena
Aranda, Carmen
Guallar, Victor
Gutiérrez Suárez, Ana
Martínez, Ángel T.
author Carro, Juan
author_facet Carro, Juan
González-Benjumea, Alejandro
Fernández-Fueyo, Elena
Aranda, Carmen
Guallar, Victor
Gutiérrez Suárez, Ana
Martínez, Ángel T.
author_role author
author2 González-Benjumea, Alejandro
Fernández-Fueyo, Elena
Aranda, Carmen
Guallar, Victor
Gutiérrez Suárez, Ana
Martínez, Ángel T.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv European Commission
Agencia Estatal de Investigación (España)
Ministerio de Economía y Competitividad (España)
Ministerio de Ciencia, Innovación y Universidades (España)
Consejo Superior de Investigaciones Científicas (España)
Carro, Juan [0000-0002-7556-9782]
González-Benjumea, Alejandro [0000-0003-2857-9491]
Fernández-Fueyo, Elena [0000-0002-5079-2240]
Aranda, Carmen [0000-0001-8213-1132]
Guallar, Victor [0000-0002-4580-1114]
Gutiérrez Suárez, Ana [0000-0002-8823-9029]
Martínez, Ángel T. [0000-0002-1584-2863]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Unspecific peroxygenase
Marasmius rotula
Unsaturated fatty acids
Epoxidation
Hydroxylation
Computational simulations
PELE software
Gas chromatography−mass spectrometry
topic Unspecific peroxygenase
Marasmius rotula
Unsaturated fatty acids
Epoxidation
Hydroxylation
Computational simulations
PELE software
Gas chromatography−mass spectrometry
description 9 p.-8 fig.-2 tab.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
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/10261/184640
url http://hdl.handle.net/10261/184640
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
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info:eu-repo/grantAgreement/EC/H2020/792063
BIO2017-86559-R/AEI/10.13039/501100011033
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2016-79138-R
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/BIO2017-86559-R
https://doi.org/10.1021/acscatal.9b01454
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
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
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spelling Modulating fatty acid epoxidation vs hydroxylation in a fungal peroxygenaseCarro, JuanGonzález-Benjumea, AlejandroFernández-Fueyo, ElenaAranda, CarmenGuallar, VictorGutiérrez Suárez, AnaMartínez, Ángel T.Unspecific peroxygenaseMarasmius rotulaUnsaturated fatty acidsEpoxidationHydroxylationComputational simulationsPELE softwareGas chromatography−mass spectrometry9 p.-8 fig.-2 tab.Unspecific peroxygenases (UPOs) are fungal secreted counterparts of the cytochrome P450 monooxygenases present in most living cells. Both enzyme types share the ability to perform selective oxygenation reactions. Moreover, the Marasmius rotula UPO (MroUPO) catalyzes reactions of interest compared with the previously described UPOs, including formation of reactive epoxy fatty acids. To investigate substrate epoxidation, the most frequent positions of oleic acid at the MroUPO heme channel were predicted using binding and molecular dynamics simulations. Then, mutations in neighbor residues were designed aiming at modulating the enzyme epoxidation vs hydroxylation ratio. Both the native (wild-type recombinant) MroUPO and the mutated variants were expressed in Escherichia coli as active enzymes, and their action on oleic and other fatty acids was investigated by gas chromatography–mass spectrometry in combination with kinetic analyses. Interestingly, a small modification of the channel shape in the I153T variant increased the ratio between epoxidized oleic acid and its additionally hydroxylated derivatives. A fully opposite effect was attained with the double I153F/S156F variant that completely abolished the ability of the MroUPO to epoxidize oleic acid (while no activity was detected for the I153V variant). The rationale for these results was revealed by the substrate positioning in the above computational simulations, which predict a shorter distance between the oleic acid double bond and the oxygen atom of the peroxide-activated heme (compound I) in the I153T variant than in the native enzyme, promoting epoxidation. In contrast, the I153F/S156F double mutation fully prevents the approach of oleic acid in the bent conformation required for double-bond epoxidation, although its (sub)terminal hydroxylation was predicted and experimentally confirmed. The I153T mutation also increased the UPO selectivity on polyunsaturated fatty acid epoxidation, strongly reducing the ratio between simple epoxides and their hydroxylated derivatives, with respect to the native UPO.This work has been funded by the BBI-JU (www.bbi-europe.eu) project SusBind (on “Sustainable biobinders”; H2020-BBI-JTI-2017-792063; https://susbind.eu), together with the CTQ2016-79138-R and BIO2017-86559-R (GENOBIOREF) projects of the Spanish Ministry of Economy, Industry and Competitiveness, cofinanced by FEDER funds.We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).Peer reviewedAmerican Chemical SocietyEuropean CommissionAgencia Estatal de Investigación (España)Ministerio de Economía y Competitividad (España)Ministerio de Ciencia, Innovación y Universidades (España)Consejo Superior de Investigaciones Científicas (España)Carro, Juan [0000-0002-7556-9782]González-Benjumea, Alejandro [0000-0003-2857-9491]Fernández-Fueyo, Elena [0000-0002-5079-2240]Aranda, Carmen [0000-0001-8213-1132]Guallar, Victor [0000-0002-4580-1114]Gutiérrez Suárez, Ana [0000-0002-8823-9029]Martínez, Ángel T. [0000-0002-1584-2863]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]201920192019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/184640reponame: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##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/EC/H2020/792063BIO2017-86559-R/AEI/10.13039/501100011033info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2016-79138-Rinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/BIO2017-86559-Rhttps://doi.org/10.1021/acscatal.9b01454Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1846402026-05-22T06:33:51Z
score 15,812455

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