Expanding the recombinant protein quality in Lactococcus lactis

Background: Escherichia coli has been a main host for the production of recombinant proteins of biomedical interest, but conformational stress responses impose severe bottlenecks that impair the production of soluble, proteolytically stable versions of many protein species. In this context, emerging...

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Autores: Cano-Garrido, Olivia|||0000-0002-5504-2131, Rueda, Fabián L., Sánchez-García, Laura|||0000-0002-8420-1701, Ruiz-Ávila, Luis, Bosser, Ramon, Villaverde, Antonio|||0000-0002-2615-4521, Garcia-Fruitos, Elena|||0000-0001-7498-4864
Tipo de recurso: artículo
Fecha de publicación:2014
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:132800
Acceso en línea:https://ddd.uab.cat/record/132800
https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3
Access Level:acceso abierto
Palabra clave:Lactococcus lactis
Solubility
Solubilitat
Recombinant protein quality
Qualitat de proteïna recombinant
Conformational quality
Qualitat conformacional
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spelling Expanding the recombinant protein quality in Lactococcus lactisCano-Garrido, Olivia|||0000-0002-5504-2131Rueda, Fabián L.Sánchez-García, Laura|||0000-0002-8420-1701Ruiz-Ávila, LuisBosser, RamonVillaverde, Antonio|||0000-0002-2615-4521Garcia-Fruitos, Elena|||0000-0001-7498-4864Lactococcus lactisSolubilitySolubilitatRecombinant protein qualityQualitat de proteïna recombinantConformational qualityQualitat conformacionalBackground: Escherichia coli has been a main host for the production of recombinant proteins of biomedical interest, but conformational stress responses impose severe bottlenecks that impair the production of soluble, proteolytically stable versions of many protein species. In this context, emerging Generally Recognized As Safe (GRAS) bacterial hosts provide alternatives as cell factories for recombinant protein production, in which limitations associated to the use of Gram-negative microorganisms might result minimized. Among them, Lactic Acid Bacteria and specially Lactococcus lactis are Gram-positive GRAS organisms in which recombinant protein solubility is generically higher and downstream facilitated, when compared to E. coli. However, deep analyses of recombinant protein quality in this system are still required to completely evaluate its performance and potential for improvement. - Results : we have explored here the conformational quality (through specific fluorescence emission) and solubility of an aggregation-prone GFP variant (VP1GFP) produced in L. lactis. In this context, our results show that parameters such as production time, culture conditions and growth temperature have a dramatic impact not only on protein yield, but also on protein solubility and conformational quality, that are particularly favored under fermentative metabolism. - Conclusions: metabolic regime and cultivation temperature greatly influence solubility and conformational quality of an aggregation-prone protein in L. lactis. Specifically, the present study proves that anaerobic growth is the optimal condition for recombinant protein production purposes. Besides, growth temperature plays an important role regulating both protein solubility and conformational quality. Additionally, our results also prove the great versatility for the manipulation of this bacterial system regarding the improvement of functionality, yield and quality of recombinant proteins in this species. These findings not only confirm L. lactis as an excellent producer of recombinant proteins but also reveal room for significant improvement by the exploitation of external protein quality modulators. 22014-01-0120142014-01-01Articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://ddd.uab.cat/record/132800https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3reponame:Dipòsit Digital de Documents de la UABinstname:Universitat Autònoma de BarcelonaInglésengMinisterio de Economía y Competitividad https://doi.org/10.13039/501100003329 RTA2012-00028-C02-02Agència de Gestió d'Ajuts Universitaris i de Recerca https://doi.org/10.13039/501100003030 2014/SGR-132open accesshttp://purl.org/coar/access_right/c_abf2Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.https://creativecommons.org/licenses/by/3.0/info:eu-repo/semantics/openAccessoai:ddd.uab.cat:1328002026-06-06T12:50:31Z
dc.title.none.fl_str_mv Expanding the recombinant protein quality in Lactococcus lactis
title Expanding the recombinant protein quality in Lactococcus lactis
spellingShingle Expanding the recombinant protein quality in Lactococcus lactis
Cano-Garrido, Olivia|||0000-0002-5504-2131
Lactococcus lactis
Solubility
Solubilitat
Recombinant protein quality
Qualitat de proteïna recombinant
Conformational quality
Qualitat conformacional
title_short Expanding the recombinant protein quality in Lactococcus lactis
title_full Expanding the recombinant protein quality in Lactococcus lactis
title_fullStr Expanding the recombinant protein quality in Lactococcus lactis
title_full_unstemmed Expanding the recombinant protein quality in Lactococcus lactis
title_sort Expanding the recombinant protein quality in Lactococcus lactis
dc.creator.none.fl_str_mv Cano-Garrido, Olivia|||0000-0002-5504-2131
Rueda, Fabián L.
Sánchez-García, Laura|||0000-0002-8420-1701
Ruiz-Ávila, Luis
Bosser, Ramon
Villaverde, Antonio|||0000-0002-2615-4521
Garcia-Fruitos, Elena|||0000-0001-7498-4864
author Cano-Garrido, Olivia|||0000-0002-5504-2131
author_facet Cano-Garrido, Olivia|||0000-0002-5504-2131
Rueda, Fabián L.
Sánchez-García, Laura|||0000-0002-8420-1701
Ruiz-Ávila, Luis
Bosser, Ramon
Villaverde, Antonio|||0000-0002-2615-4521
Garcia-Fruitos, Elena|||0000-0001-7498-4864
author_role author
author2 Rueda, Fabián L.
Sánchez-García, Laura|||0000-0002-8420-1701
Ruiz-Ávila, Luis
Bosser, Ramon
Villaverde, Antonio|||0000-0002-2615-4521
Garcia-Fruitos, Elena|||0000-0001-7498-4864
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Lactococcus lactis
Solubility
Solubilitat
Recombinant protein quality
Qualitat de proteïna recombinant
Conformational quality
Qualitat conformacional
topic Lactococcus lactis
Solubility
Solubilitat
Recombinant protein quality
Qualitat de proteïna recombinant
Conformational quality
Qualitat conformacional
description Background: Escherichia coli has been a main host for the production of recombinant proteins of biomedical interest, but conformational stress responses impose severe bottlenecks that impair the production of soluble, proteolytically stable versions of many protein species. In this context, emerging Generally Recognized As Safe (GRAS) bacterial hosts provide alternatives as cell factories for recombinant protein production, in which limitations associated to the use of Gram-negative microorganisms might result minimized. Among them, Lactic Acid Bacteria and specially Lactococcus lactis are Gram-positive GRAS organisms in which recombinant protein solubility is generically higher and downstream facilitated, when compared to E. coli. However, deep analyses of recombinant protein quality in this system are still required to completely evaluate its performance and potential for improvement. - Results : we have explored here the conformational quality (through specific fluorescence emission) and solubility of an aggregation-prone GFP variant (VP1GFP) produced in L. lactis. In this context, our results show that parameters such as production time, culture conditions and growth temperature have a dramatic impact not only on protein yield, but also on protein solubility and conformational quality, that are particularly favored under fermentative metabolism. - Conclusions: metabolic regime and cultivation temperature greatly influence solubility and conformational quality of an aggregation-prone protein in L. lactis. Specifically, the present study proves that anaerobic growth is the optimal condition for recombinant protein production purposes. Besides, growth temperature plays an important role regulating both protein solubility and conformational quality. Additionally, our results also prove the great versatility for the manipulation of this bacterial system regarding the improvement of functionality, yield and quality of recombinant proteins in this species. These findings not only confirm L. lactis as an excellent producer of recombinant proteins but also reveal room for significant improvement by the exploitation of external protein quality modulators.
publishDate 2014
dc.date.none.fl_str_mv 2
2014-01-01
2014
2014-01-01
dc.type.none.fl_str_mv Article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://ddd.uab.cat/record/132800
https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3
url https://ddd.uab.cat/record/132800
https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Economía y Competitividad https://doi.org/10.13039/501100003329 RTA2012-00028-C02-02
Agència de Gestió d'Ajuts Universitaris i de Recerca https://doi.org/10.13039/501100003030 2014/SGR-132
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
https://creativecommons.org/licenses/by/3.0/
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
https://creativecommons.org/licenses/by/3.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Dipòsit Digital de Documents de la UAB
instname:Universitat Autònoma de Barcelona
instname_str Universitat Autònoma de Barcelona
reponame_str Dipòsit Digital de Documents de la UAB
collection Dipòsit Digital de Documents de la UAB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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