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...
| Autores: | ; ; ; ; ; ; |
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| Tipo de documento: | Artigo |
| Data de publicação: | 2014 |
| País: | España |
| Recursos: | Universitat Autònoma de Barcelona |
| Repositório: | Dipòsit Digital de Documents de la UAB |
| Idioma: | Inglês |
| OAI Identifier: | oai:ddd.uab.cat:132800 |
| Acesso em linha: | https://ddd.uab.cat/record/132800 https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3 |
| Nível de acesso: | Acesso aberto |
| Palavra-chave: | Lactococcus lactis Solubility Solubilitat Recombinant protein quality Qualitat de proteïna recombinant Conformational quality Qualitat conformacional |
| _version_ | 1878730383449653248 |
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| author | Cano-Garrido, Olivia|||0000-0002-5504-2131 |
| 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 |
| author_browse | Bosser, Ramon Cano-Garrido, Olivia|||0000-0002-5504-2131 Garcia-Fruitos, Elena|||0000-0001-7498-4864 Rueda, Fabián L. Ruiz-Ávila, Luis Sánchez-García, Laura|||0000-0002-8420-1701 Villaverde, Antonio|||0000-0002-2615-4521 |
| 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 |
| collection | Dipòsit Digital de Documents de la UAB |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_1b39f2f6330eef0b5d8f33652f8f8dfd |
| institution | Universitat Autònoma de Barcelona |
| instname_str | Universitat Autònoma de Barcelona |
| language | eng |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:ddd.uab.cat:132800 |
| publishDate | 2014 |
| publishDateSort | 2014 |
| reponame_str | Dipòsit Digital de Documents de la UAB |
| spelling | Expanding the recombinant protein quality in Lactococcus lactis 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 Lactococcus lactis Solubility Solubilitat Recombinant protein quality Qualitat de proteïna recombinant Conformational quality Qualitat conformacional 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. https://ddd.uab.cat/record/132800 https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3 |
| 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 | 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_short | Expanding the recombinant protein quality in Lactococcus lactis |
| title_sort | Expanding the recombinant protein quality in Lactococcus lactis |
| topic | Lactococcus lactis Solubility Solubilitat Recombinant protein quality Qualitat de proteïna recombinant Conformational quality Qualitat conformacional |
| topic_facet | Lactococcus lactis Solubility Solubilitat Recombinant protein quality Qualitat de proteïna recombinant Conformational quality Qualitat conformacional |
| url | https://ddd.uab.cat/record/132800 https://dx.doi.org/urn:doi:10.1186/s12934-014-0167-3 |