Stable expression plasmids for Streptomyces based on a toxin-antitoxin system

This is an Open Access article distributed under the terms of the Creative Commons Attribution License.

Detalles Bibliográficos
Autores: Sevillano, Laura, Díaz, Margarita, Santamaría, Ramón I.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2013
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/76116
Acceso en línea:http://hdl.handle.net/10261/76116
Access Level:acceso abierto
Palabra clave:Streptomyces
Toxin-antitoxin
Recombinant protein
Plasmid stabilization
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spelling Stable expression plasmids for Streptomyces based on a toxin-antitoxin systemSevillano, LauraDíaz, MargaritaSantamaría, Ramón I.StreptomycesToxin-antitoxinRecombinant proteinPlasmid stabilizationThis is an Open Access article distributed under the terms of the Creative Commons Attribution License.[Background]: Bacteria included in the genus Streptomyces exhibit several attractive characteristics that make them adequate hosts for the heterologous expression of proteins. One of them is that some of its species have a high secretion capacity and hence the protein of interest could be released to the culture supernatant, facilitating downstream processing. To date, all the expression vectors described for these bacteria contain antibiotic resistance genes as selection markers. However, the use of antibiotics to produce proteins at industrial level is currently becoming more restricted owing to the possibility of contamination of the final product. In this report, we describe the use of the S. lividans yefM/yoeBsl toxin-antitoxin system to develop a stable plasmid expression system. [Results]: In order to use the yefM/yoeBsl system to stabilize expression plasmids in Streptomyces, a S. lividans mutant strain that contained only the toxin gene (yoeBsl) in its genome and the antitoxin gene (yefMsl) located in a temperature-sensitive plasmid was constructed and used as host. This strain was transformed with an expression plasmid harbouring both the antitoxin gene and the gene encoding the protein of interest. Thus, after elimination of the temperature-sensitive plasmid, only cells with the expression plasmid were able to survive. On using this system, two proteins - an α-amylase from S. griseus and a xylanase from S. halstedii - were overproduced without the addition of antibiotic to the culture medium. The production of both proteins was high, even after long incubations (8 days), and after serial subcultures, confirming the stability of the plasmids without antibiotic selection. [Conclusions]: This is the first report that describes the use of a toxin-antitoxin system to maintain high -copy plasmids in Streptomyces. This finding could be a valuable tool for using Streptomyces as a host to produce proteins at the industrial and pharmaceutical levels without the use of antibiotics in the production step.Research in our lab is supported by the grants EUI2008-03631 and BFU2010-17551 from the Ministerio de Ciencia e Innovación to R. Santamaría and M. Díaz respectively and by the grant CSI099A12-1 from the Junta de Castilla y León to R. Santamaría. The authors also thank the CSIC for the co-financing of this publication in Open Access. The IBFG acknowledges the institutional support granted by the Ramón Areces Foundation during 2011– 2012.The authors also thank the Unit of Information Resources for Research (URICI-CSIC) for the co-financing of this publication in Open Access.Peer ReviewedBioMed CentralCSIC - Unidad de Recursos de Información Científica para la Investigación (URICI)Ministerio de Ciencia e Innovación (España)Consejo Superior de Investigaciones Científicas (España)Fundación Ramón ArecesJunta de Castilla y LeónConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2013201320132013info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/76116reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1186/1475-2859-12-39Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/761162026-05-22T06:33:51Z
dc.title.none.fl_str_mv Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
title Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
spellingShingle Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
Sevillano, Laura
Streptomyces
Toxin-antitoxin
Recombinant protein
Plasmid stabilization
title_short Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
title_full Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
title_fullStr Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
title_full_unstemmed Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
title_sort Stable expression plasmids for Streptomyces based on a toxin-antitoxin system
dc.creator.none.fl_str_mv Sevillano, Laura
Díaz, Margarita
Santamaría, Ramón I.
author Sevillano, Laura
author_facet Sevillano, Laura
Díaz, Margarita
Santamaría, Ramón I.
author_role author
author2 Díaz, Margarita
Santamaría, Ramón I.
author2_role author
author
dc.contributor.none.fl_str_mv CSIC - Unidad de Recursos de Información Científica para la Investigación (URICI)
Ministerio de Ciencia e Innovación (España)
Consejo Superior de Investigaciones Científicas (España)
Fundación Ramón Areces
Junta de Castilla y León
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Streptomyces
Toxin-antitoxin
Recombinant protein
Plasmid stabilization
topic Streptomyces
Toxin-antitoxin
Recombinant protein
Plasmid stabilization
description This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
publishDate 2013
dc.date.none.fl_str_mv 2013
2013
2013
2013
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/10261/76116
url http://hdl.handle.net/10261/76116
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1186/1475-2859-12-39
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
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 15.81155

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