Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis

[EN] The oppA2 gene encodes an oligopeptide-binding protein similar to the periplasmic substrate-binding proteins of the ABC transport systems. However, oppA2 is an orphan gene, not included in an ABC operon. This gene is located in the clavulanic acid (CA) gene cluster of Streptomyces clavuligerus...

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Autores: Álvarez Álvarez, Rubén, Rodríguez García, Antonio, Martínez Burgo, Yolanda, Martín Martín, Juan Francisco, Liras Padín, Paloma
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2018
País:España
Institución:Universidad Rey Juan Carlos
Repositorio:BULERIA. Repositorio Institucional de la Universidad de León
OAI Identifier:oai:buleria.unileon.es:10612/18073
Acceso en línea:https://journals.asm.org/doi/10.1128/aem.01701-18
https://hdl.handle.net/10612/18073
Access Level:acceso abierto
Palabra clave:Biotecnología
Streptomyces clavuligerus
Oligopeptide-binding protein OppA2
oppA2-null mutant
Antibotics biosynthesis regulation
Clavulanic acid gene cluster
Transcriptomics
2414.01 Antibióticos
2415.01 Biología Molecular de Microorganismos
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oai_identifier_str oai:buleria.unileon.es:10612/18073
network_acronym_str ES
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repository_id_str
spelling Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesisÁlvarez Álvarez, RubénRodríguez García, AntonioMartínez Burgo, YolandaMartín Martín, Juan FranciscoLiras Padín, PalomaBiotecnologíaStreptomyces clavuligerusOligopeptide-binding protein OppA2oppA2-null mutantAntibotics biosynthesis regulationClavulanic acid gene clusterTranscriptomics2414.01 Antibióticos2415.01 Biología Molecular de Microorganismos[EN] The oppA2 gene encodes an oligopeptide-binding protein similar to the periplasmic substrate-binding proteins of the ABC transport systems. However, oppA2 is an orphan gene, not included in an ABC operon. This gene is located in the clavulanic acid (CA) gene cluster of Streptomyces clavuligerus and is essential for CA production. A transcriptomic study of the oppA2-null mutant S. clavuligerus ΔoppA2::aac showed changes in the expression levels of 233 genes from those in the parental strain. These include genes for ABC transport systems, secreted proteins, peptidases, and proteases. Expression of the clavulanic acid, clavam, and cephamycin C biosynthesis gene clusters was not significantly affected in the oppA2 deletion mutant. The genes for holomycin biosynthesis were upregulated 2-fold on average, and the level of upregulation increased to 43-fold in a double mutant lacking oppA2 and the pSCL4 plasmid. Strains in which oppA2 was mutated secreted into the culture the compound N-acetylglycyl-clavaminic acid (AGCA), a putative intermediate of CA biosynthesis. A culture broth containing AGCA, or AGCA purified by liquid chromatography-mass spectrometry (LC-MS), was added to the cultures of various non-CA-producing mutants. Mutants blocked in the early steps of the pathway restored CA production, whereas mutants altered in late steps did not, establishing that AGCA is a late intermediate of the biosynthetic pathway, which is released from the cells when the oligopeptide-binding protein OppA2 is not availableSIThis work was supported by grant BIO2013-34723 from the Spanish Ministry of Economy and Competitiveness. R. Álvarez-Álvarez and Y. Martínez-Burgo received a PFU fellowship from the Spanish Ministry of Science and InnovationAmerican Society for Microbiology (ASM)MicrobiologiaFacultad de Ciencias Biologicas y Ambientales2018info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://journals.asm.org/doi/10.1128/aem.01701-18https://hdl.handle.net/10612/18073reponame:BULERIA. Repositorio Institucional de la Universidad de Leóninstname:Universidad Rey Juan CarlosInglésinfo:eu-repo/grantAgreement/MINECO//BIO2013-34723info:eu-repo/semantics/openAccessoai:buleria.unileon.es:10612/180732026-06-24T12:43:27Z
dc.title.none.fl_str_mv Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
title Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
spellingShingle Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
Álvarez Álvarez, Rubén
Biotecnología
Streptomyces clavuligerus
Oligopeptide-binding protein OppA2
oppA2-null mutant
Antibotics biosynthesis regulation
Clavulanic acid gene cluster
Transcriptomics
2414.01 Antibióticos
2415.01 Biología Molecular de Microorganismos
title_short Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
title_full Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
title_fullStr Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
title_full_unstemmed Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
title_sort Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
dc.creator.none.fl_str_mv Álvarez Álvarez, Rubén
Rodríguez García, Antonio
Martínez Burgo, Yolanda
Martín Martín, Juan Francisco
Liras Padín, Paloma
author Álvarez Álvarez, Rubén
author_facet Álvarez Álvarez, Rubén
Rodríguez García, Antonio
Martínez Burgo, Yolanda
Martín Martín, Juan Francisco
Liras Padín, Paloma
author_role author
author2 Rodríguez García, Antonio
Martínez Burgo, Yolanda
Martín Martín, Juan Francisco
Liras Padín, Paloma
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Microbiologia
Facultad de Ciencias Biologicas y Ambientales
dc.subject.none.fl_str_mv Biotecnología
Streptomyces clavuligerus
Oligopeptide-binding protein OppA2
oppA2-null mutant
Antibotics biosynthesis regulation
Clavulanic acid gene cluster
Transcriptomics
2414.01 Antibióticos
2415.01 Biología Molecular de Microorganismos
topic Biotecnología
Streptomyces clavuligerus
Oligopeptide-binding protein OppA2
oppA2-null mutant
Antibotics biosynthesis regulation
Clavulanic acid gene cluster
Transcriptomics
2414.01 Antibióticos
2415.01 Biología Molecular de Microorganismos
description [EN] The oppA2 gene encodes an oligopeptide-binding protein similar to the periplasmic substrate-binding proteins of the ABC transport systems. However, oppA2 is an orphan gene, not included in an ABC operon. This gene is located in the clavulanic acid (CA) gene cluster of Streptomyces clavuligerus and is essential for CA production. A transcriptomic study of the oppA2-null mutant S. clavuligerus ΔoppA2::aac showed changes in the expression levels of 233 genes from those in the parental strain. These include genes for ABC transport systems, secreted proteins, peptidases, and proteases. Expression of the clavulanic acid, clavam, and cephamycin C biosynthesis gene clusters was not significantly affected in the oppA2 deletion mutant. The genes for holomycin biosynthesis were upregulated 2-fold on average, and the level of upregulation increased to 43-fold in a double mutant lacking oppA2 and the pSCL4 plasmid. Strains in which oppA2 was mutated secreted into the culture the compound N-acetylglycyl-clavaminic acid (AGCA), a putative intermediate of CA biosynthesis. A culture broth containing AGCA, or AGCA purified by liquid chromatography-mass spectrometry (LC-MS), was added to the cultures of various non-CA-producing mutants. Mutants blocked in the early steps of the pathway restored CA production, whereas mutants altered in late steps did not, establishing that AGCA is a late intermediate of the biosynthetic pathway, which is released from the cells when the oligopeptide-binding protein OppA2 is not available
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://journals.asm.org/doi/10.1128/aem.01701-18
https://hdl.handle.net/10612/18073
url https://journals.asm.org/doi/10.1128/aem.01701-18
https://hdl.handle.net/10612/18073
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/MINECO//BIO2013-34723
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 (ASM)
publisher.none.fl_str_mv American Society for Microbiology (ASM)
dc.source.none.fl_str_mv reponame:BULERIA. Repositorio Institucional de la Universidad de León
instname:Universidad Rey Juan Carlos
instname_str Universidad Rey Juan Carlos
reponame_str BULERIA. Repositorio Institucional de la Universidad de León
collection BULERIA. Repositorio Institucional de la Universidad de León
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.812429

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