Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope

17 pags., 7 figs., 1 tab.

Detalles Bibliográficos
Autores: Cea-Rama, Isabel, Coscolín, Cristina, González-Alfonso, José L., Raj, Jog, Vasiljević, Marko, Plou Gasca, Francisco José, Ferrer, Manuel, Sanz-Aparicio, J.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/289557
Acceso en línea:http://hdl.handle.net/10261/289557
https://api.elsevier.com/content/abstract/scopus_id/85132755832
Access Level:acceso abierto
Palabra clave:Esterase
Metagenome
Promiscuity
α/β-fold hydrolase
β-lactamase
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dc.title.none.fl_str_mv Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
title Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
spellingShingle Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
Cea-Rama, Isabel
Esterase
Metagenome
Promiscuity
α/β-fold hydrolase
β-lactamase
title_short Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
title_full Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
title_fullStr Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
title_full_unstemmed Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
title_sort Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope
dc.creator.none.fl_str_mv Cea-Rama, Isabel
Coscolín, Cristina
González-Alfonso, José L.
Raj, Jog
Vasiljević, Marko
Plou Gasca, Francisco José
Ferrer, Manuel
Sanz-Aparicio, J.
author Cea-Rama, Isabel
author_facet Cea-Rama, Isabel
Coscolín, Cristina
González-Alfonso, José L.
Raj, Jog
Vasiljević, Marko
Plou Gasca, Francisco José
Ferrer, Manuel
Sanz-Aparicio, J.
author_role author
author2 Coscolín, Cristina
González-Alfonso, José L.
Raj, Jog
Vasiljević, Marko
Plou Gasca, Francisco José
Ferrer, Manuel
Sanz-Aparicio, J.
author2_role author
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 e Innovación (España)
Consejo Superior de Investigaciones Científicas (España)
Ministerio de Educación, Cultura y Deporte (España)
ALBA Synchrotron
Ferrer, Manuel [0000-0003-4962-4714]
Sanz-Aparicio, J. [0000-0002-6849-8621]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Esterase
Metagenome
Promiscuity
α/β-fold hydrolase
β-lactamase
topic Esterase
Metagenome
Promiscuity
α/β-fold hydrolase
β-lactamase
description 17 pags., 7 figs., 1 tab.
publishDate 2022
dc.date.none.fl_str_mv 2022
2023
2023
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/10261/289557
https://api.elsevier.com/content/abstract/scopus_id/85132755832
url http://hdl.handle.net/10261/289557
https://api.elsevier.com/content/abstract/scopus_id/85132755832
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
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info:eu-repo/grantAgreement/EC/H2020/101000327
info:eu-repo/grantAgreement/EC/FP7/604814
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-85522-R
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-112758RB-I00
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info:eu-repo/grantAgreement/MINECO//BES-2015-073829
The FEBS journal
https://doi.org/10.1111/febs.16554
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Federation of European Biochemical Societies
publisher.none.fl_str_mv Federation of European Biochemical Societies
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
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spelling Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scopeCea-Rama, IsabelCoscolín, CristinaGonzález-Alfonso, José L.Raj, JogVasiljević, MarkoPlou Gasca, Francisco JoséFerrer, ManuelSanz-Aparicio, J.EsteraseMetagenomePromiscuityα/β-fold hydrolaseβ-lactamase17 pags., 7 figs., 1 tab.Family VIII esterases present similarities to class C β-lactamases, which show nucleophilic serines located at the S-X-X-K motif instead of the G-X-S-X-G or G-D-S-(L) motif shown by other carboxylesterase families. Here, we report the crystal structure of a novel family VIII (subfamily VIII. I) esterase (EH7 ; denaturing temperature, 52.6 ± 0.3 °C; pH optimum 7.0-9.0) to deepen its broad substrate range. Indeed, the analysis of the substrate specificity revealed its capacity to hydrolyse nitrocefin as a model chromogenic cephalosporin substrate (40.4 ± 11.4 units·g-1 ), and a large battery of 66 structurally different esters (up to 1730 min-1 ), including bis(2-hydroxyethyl)-terephthalate (241.7 ± 8.5 units·g-1 ) and the mycotoxin T-2 (1220 ± 52 units·g-1 ). It also showed acyltransferase activity through the synthesis of benzyl 3-oxobutanoate (40.4 ± 11.4 units·g-1 ) from benzyl alcohol and vinyl acetoacetate. Such a broad substrate scope is rare among family VIII esterases and lipolytic enzymes. Structural analyses of free and substrate-bound forms of this homooctamer esterase suggest that EH7 presents a more opened and exposed S1 site having no steric hindrance for the entrance of substrates to the active site, more flexible R1, R2 and R3 regions allowing for the binding of a wide spectrum of substrates into the active site, and small residues in the conserved motif Y-X-X containing the catalytic Tyr enabling the entrance of large substrates. These unique structural elements in combination with docking experiments allowed us to gain valuable insights into the substrate specificity of this esterase and possible others belonging to family VIII.This study was conducted under the auspices of the FuturEnzyme Project funded by the European Union's Horizon 2020 Research and Innovation Programme under Grant Agreement No. 101000327. We also acknowledge financial support under Grants PCIN-2017-078 (within the Marine Biotechnology ERA-NET, GA No. 604814), BIO2017-85522-R (MF), PID2020-112758RB-I00 (MF), PDC2021-121534-I00 (MF), PID2019-105838RB-C31 (FJP) and PID2019-105838RB-C33 (JS-A) from the Ministerio de Economía, Industria y Competitividad, Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (AEI) (Digital Object Identifier 10.13039/501100011033), Fondo Europeo de Desarrollo Regional (FEDER) and the European Union (‘NextGen-erationEU/PRTR’), and Grant 2020AEP061 (MF) from the Agencia Estatal CSIC. C Coscolín thanks the Ministerio de Economía y Competitividad and FEDER for a PhD fellowship (Grant BES-2015-073829). JLG-A acknowledges support from the Spanish Ministry of Education, Culture and Sport through the National Program FPU (FPU17/00044). The authors acknowledge David Almendral and Rutz Matesanz for supporting the circular dichroism analysis, and Rafael Bargiela for the design of Fig. 2. We thank the staff of the Synchrotron Radiation Source at Alba (Barcelona, Spain) for assistance at the BL13-XALOC beamline. Open access funding enabled and organized by ProjektDEAL.Peer reviewedFederation of European Biochemical SocietiesEuropean CommissionAgencia Estatal de Investigación (España)Ministerio de Economía y Competitividad (España)Ministerio de Ciencia e Innovación (España)Consejo Superior de Investigaciones Científicas (España)Ministerio de Educación, Cultura y Deporte (España)ALBA SynchrotronFerrer, Manuel [0000-0003-4962-4714]Sanz-Aparicio, J. [0000-0002-6849-8621]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202320232022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/289557https://api.elsevier.com/content/abstract/scopus_id/85132755832reponame:DIGITAL.CSIC. 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