A new l-proline amide hydrolase with potential application within the amidase process

L-proline amide hydrolase (PAH, EC 3.5.1.101) is a barely described enzyme belonging to the peptidase S33 family, and is highly similar to prolyl aminopeptidases (PAP, EC. 3.4.11.5). Besides being an S-stereoselective character towards piperidine-based carboxamides, this enzyme also hydrolyses diffe...

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Detalles Bibliográficos
Autores: Martínez-Rodríguez, S., Contreras-Montoya, Rafael, Torres, Juan María, Cienfuegos Rodríguez, Luis Álvarez de, Gavira Gallardo, J. A.
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:10261/334018
Acceso en línea:http://hdl.handle.net/10261/334018
Access Level:acceso abierto
Palabra clave:Amidase
Amidase process
Amino acid
Aminopeptidase
Proline
S33 family
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spelling A new l-proline amide hydrolase with potential application within the amidase processMartínez-Rodríguez, S.Contreras-Montoya, RafaelTorres, Juan MaríaCienfuegos Rodríguez, Luis Álvarez deGavira Gallardo, J. A.AmidaseAmidase processAmino acidAminopeptidaseProlineS33 familyL-proline amide hydrolase (PAH, EC 3.5.1.101) is a barely described enzyme belonging to the peptidase S33 family, and is highly similar to prolyl aminopeptidases (PAP, EC. 3.4.11.5). Besides being an S-stereoselective character towards piperidine-based carboxamides, this enzyme also hydrolyses different L-amino acid amides, turning it into a potential biocatalyst within the Amidase Process. In this work, we report the characterization of L-proline amide hydrolase from Pseudomonas syringae (PsyPAH) together with the first X-ray structure for this class of L-amino acid amidases. Recombinant PsyPAH showed optimal conditions at pH 7.0 and 35 C, with an apparent thermal melting temperature of 46 C. The enzyme behaved as a monomer at the optimal pH. The L-enantioselective hydrolytic activity towards different canonical and non-canonical amino-acid amides was confirmed. Structural analysis suggests key residues in the enzymatic activity.This research was supported by the Spanish Ministry of Science and Innovation/FEDER funds grant PID2020-116261GB-I00/AEI/10.13039/501100011033 (JAG), from the FEDER/Junta de Andalucía-Consejería de Transformación Económica, Industria, Conocimiento y Universidades grants P18-FR-3533 (LAC) and P12-FQM-790 (RCM), and from the University of Granada grant PPJI2017-1 (SMR).Multidisciplinary Digital Publishing InstituteMinisterio de Ciencia e Innovación (España)Junta de AndalucíaUniversidad de GranadaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2023202320222023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/334018reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO//PID2020-116261GB-I00http://dx.doi.org/10.3390/cryst12010018Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3340182026-05-22T06:33:51Z
dc.title.none.fl_str_mv A new l-proline amide hydrolase with potential application within the amidase process
title A new l-proline amide hydrolase with potential application within the amidase process
spellingShingle A new l-proline amide hydrolase with potential application within the amidase process
Martínez-Rodríguez, S.
Amidase
Amidase process
Amino acid
Aminopeptidase
Proline
S33 family
title_short A new l-proline amide hydrolase with potential application within the amidase process
title_full A new l-proline amide hydrolase with potential application within the amidase process
title_fullStr A new l-proline amide hydrolase with potential application within the amidase process
title_full_unstemmed A new l-proline amide hydrolase with potential application within the amidase process
title_sort A new l-proline amide hydrolase with potential application within the amidase process
dc.creator.none.fl_str_mv Martínez-Rodríguez, S.
Contreras-Montoya, Rafael
Torres, Juan María
Cienfuegos Rodríguez, Luis Álvarez de
Gavira Gallardo, J. A.
author Martínez-Rodríguez, S.
author_facet Martínez-Rodríguez, S.
Contreras-Montoya, Rafael
Torres, Juan María
Cienfuegos Rodríguez, Luis Álvarez de
Gavira Gallardo, J. A.
author_role author
author2 Contreras-Montoya, Rafael
Torres, Juan María
Cienfuegos Rodríguez, Luis Álvarez de
Gavira Gallardo, J. A.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (España)
Junta de Andalucía
Universidad de Granada
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Amidase
Amidase process
Amino acid
Aminopeptidase
Proline
S33 family
topic Amidase
Amidase process
Amino acid
Aminopeptidase
Proline
S33 family
description L-proline amide hydrolase (PAH, EC 3.5.1.101) is a barely described enzyme belonging to the peptidase S33 family, and is highly similar to prolyl aminopeptidases (PAP, EC. 3.4.11.5). Besides being an S-stereoselective character towards piperidine-based carboxamides, this enzyme also hydrolyses different L-amino acid amides, turning it into a potential biocatalyst within the Amidase Process. In this work, we report the characterization of L-proline amide hydrolase from Pseudomonas syringae (PsyPAH) together with the first X-ray structure for this class of L-amino acid amidases. Recombinant PsyPAH showed optimal conditions at pH 7.0 and 35 C, with an apparent thermal melting temperature of 46 C. The enzyme behaved as a monomer at the optimal pH. The L-enantioselective hydrolytic activity towards different canonical and non-canonical amino-acid amides was confirmed. Structural analysis suggests key residues in the enzymatic activity.
publishDate 2022
dc.date.none.fl_str_mv 2022
2023
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/10261/334018
url http://hdl.handle.net/10261/334018
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/MINECO//PID2020-116261GB-I00
http://dx.doi.org/10.3390/cryst12010018
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
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 15,812429

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