Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues

The pva gene from Streptomyces lavendulae ATCC 13664, encoding a novel penicillin V acylase (SlPVA), has been isolated and characterized. The gene encodes an inactive precursor protein containing a secretion signal peptide that is activated by two internal autoproteolytic cleavages that release a 25...

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Detalles Bibliográficos
Autores: Torres Bacete, Jesús, Hormigo, Daniel, Torres Guzmán, Raquel, Arroyo Sánchez, Miguel, Castillón, María Pilar, García, José Luis, Acebal Sarabia, Carmen, De La Mata Riesco, Mª Isabel
Tipo de recurso: artículo
Fecha de publicación:2015
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/23383
Acceso en línea:https://hdl.handle.net/20.500.14352/23383
Access Level:acceso abierto
Palabra clave:577.2
579
577.15
Microbiology
Penicillin V Acylase
Streptomyces lavendulae
Biología
Biología molecular (Biología)
Microbiología (Biología)
24 Ciencias de la Vida
2415 Biología Molecular
2414 Microbiología
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oai_identifier_str oai:docta.ucm.es:20.500.14352/23383
network_acronym_str ES
network_name_str España
repository_id_str
spelling Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic ResiduesTorres Bacete, JesúsHormigo, DanielTorres Guzmán, RaquelArroyo Sánchez, MiguelCastillón, María PilarGarcía, José LuisAcebal Sarabia, CarmenDe La Mata Riesco, Mª Isabel577.2579577.15MicrobiologyPenicillin V AcylaseStreptomyces lavendulaeBiologíaBiología molecular (Biología)Microbiología (Biología)24 Ciencias de la Vida2415 Biología Molecular2414 MicrobiologíaThe pva gene from Streptomyces lavendulae ATCC 13664, encoding a novel penicillin V acylase (SlPVA), has been isolated and characterized. The gene encodes an inactive precursor protein containing a secretion signal peptide that is activated by two internal autoproteolytic cleavages that release a 25-amino-acid linker peptide and two large domains of 18.79 kDa (_-subunit) and 60.09 kDa (_-subunit). Based on sequence alignments and the three-dimensional model of SlPVA, the enzyme contains a hydrophobic pocket involved in catalytic activity, including Ser_1, His_23, Val_70, and Asn_272, which were confirmed by site-directed mutagenesis studies. The heterologous expression of pva in S. lividans led to the production of an extracellularly homogeneous heterodimeric enzyme at a 5-fold higher concentration (959 IU/liter) than in the original host and in a considerably shorter time. According to the catalytic properties of SlPVA, the enzyme must be classified as a new member of the Ntn-hydrolase superfamily, which belongs to a novel subfamily of acylases that recognize substrates with long hydrophobic acyl chains and have biotechnological applications in semisynthetic antifungal production.ASMUniversidad Complutense de Madrid20152015-02-0120152015-02-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/23383reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Atribución 3.0 Españahttps://creativecommons.org/licenses/by/3.0/es/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/233832026-06-02T12:44:21Z
dc.title.none.fl_str_mv Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
title Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
spellingShingle Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
Torres Bacete, Jesús
577.2
579
577.15
Microbiology
Penicillin V Acylase
Streptomyces lavendulae
Biología
Biología molecular (Biología)
Microbiología (Biología)
24 Ciencias de la Vida
2415 Biología Molecular
2414 Microbiología
title_short Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
title_full Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
title_fullStr Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
title_full_unstemmed Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
title_sort Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
dc.creator.none.fl_str_mv Torres Bacete, Jesús
Hormigo, Daniel
Torres Guzmán, Raquel
Arroyo Sánchez, Miguel
Castillón, María Pilar
García, José Luis
Acebal Sarabia, Carmen
De La Mata Riesco, Mª Isabel
author Torres Bacete, Jesús
author_facet Torres Bacete, Jesús
Hormigo, Daniel
Torres Guzmán, Raquel
Arroyo Sánchez, Miguel
Castillón, María Pilar
García, José Luis
Acebal Sarabia, Carmen
De La Mata Riesco, Mª Isabel
author_role author
author2 Hormigo, Daniel
Torres Guzmán, Raquel
Arroyo Sánchez, Miguel
Castillón, María Pilar
García, José Luis
Acebal Sarabia, Carmen
De La Mata Riesco, Mª Isabel
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv 577.2
579
577.15
Microbiology
Penicillin V Acylase
Streptomyces lavendulae
Biología
Biología molecular (Biología)
Microbiología (Biología)
24 Ciencias de la Vida
2415 Biología Molecular
2414 Microbiología
topic 577.2
579
577.15
Microbiology
Penicillin V Acylase
Streptomyces lavendulae
Biología
Biología molecular (Biología)
Microbiología (Biología)
24 Ciencias de la Vida
2415 Biología Molecular
2414 Microbiología
description The pva gene from Streptomyces lavendulae ATCC 13664, encoding a novel penicillin V acylase (SlPVA), has been isolated and characterized. The gene encodes an inactive precursor protein containing a secretion signal peptide that is activated by two internal autoproteolytic cleavages that release a 25-amino-acid linker peptide and two large domains of 18.79 kDa (_-subunit) and 60.09 kDa (_-subunit). Based on sequence alignments and the three-dimensional model of SlPVA, the enzyme contains a hydrophobic pocket involved in catalytic activity, including Ser_1, His_23, Val_70, and Asn_272, which were confirmed by site-directed mutagenesis studies. The heterologous expression of pva in S. lividans led to the production of an extracellularly homogeneous heterodimeric enzyme at a 5-fold higher concentration (959 IU/liter) than in the original host and in a considerably shorter time. According to the catalytic properties of SlPVA, the enzyme must be classified as a new member of the Ntn-hydrolase superfamily, which belongs to a novel subfamily of acylases that recognize substrates with long hydrophobic acyl chains and have biotechnological applications in semisynthetic antifungal production.
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-02-01
2015
2015-02-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.14352/23383
url https://hdl.handle.net/20.500.14352/23383
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 3.0 España
https://creativecommons.org/licenses/by/3.0/es/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 3.0 España
https://creativecommons.org/licenses/by/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv ASM
publisher.none.fl_str_mv ASM
dc.source.none.fl_str_mv reponame:Docta Complutense
instname:Universidad Complutense de Madrid (UCM)
instname_str Universidad Complutense de Madrid (UCM)
reponame_str Docta Complutense
collection Docta Complutense
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.300724

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