Web of Science: 6 citations, Scopus: 7 citations, Google Scholar: citations
Co-production of GroELS discriminates between intrinsic and thermally-induced recombinant protein aggregation during substrate quality control
Platas, Gemma (Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Rodríguez-Carmona, Escarlata (Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
García Fruitós, Elena (CIBER de Bioingeniería, Biomateriales y Nanomedicina)
Cano Garrido, Olivia (Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Villaverde Corrales, Antonio (Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Universitat Autònoma de Barcelona. Departament de Genètica i Microbiologia

Date: 2011
Abstract: The effects and effectiveness of the chaperone pair GroELS on the yield and quality of recombinant polypeptides produced in Escherichia coli are matter of controversy, as the reported activities of this complex are not always consistent and eventually indicate undesired side effects. The divergence in the reported data could be due, at least partially, to different experimental conditions in independent research approaches. We have then selected two structurally different model proteins (namely GFP and E. coli β-galactosidase) and two derived aggregation-prone fusions to explore, in a systematic way, the eventual effects of GroELS co-production on yield, solubility and conformational quality. Host cells were cultured at two alternative temperatures below the threshold at which thermal stress is expected to be triggered, to minimize the involvement of independent stress factors. From the analysis of protein yield, solubility and biological activity of the four model proteins produced alone or along the chaperones, we conclude that GroELS impacts on yield and quality of aggregation-prone proteins with intrinsic determinants but not on thermally induced protein aggregation. No effective modifications of protein solubility have been observed, but significant stabilization of small (encapsulable) substrates and moderate chaperone-induced degradation of larger (excluded) polypeptides. These findings indicate that the activities of this chaperone pair in the context of actively producing recombinant bacteria discriminate between intrinsic and thermally-induced protein aggregation, and that the side effects of GroELS overproduction might be determined by substrate size.
Note: Altres ajuts: ERANET-UB98-007
Note: Número d'acord de subvenció MICINN/BFU2010/17450
Note: Número d'acord de subvenció MICINN/ACI2009/0919
Note: Número d'acord de subvenció MICINN/EUI2008/03610
Note: Número d'acord de subvenció AGAUR/2009/SGR-108
Rights: Aquest 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. Creative Commons
Language: Anglès.
Document: article ; recerca ; publishedVersion
Subject: Protein Yield ; Recombinant Protein Production ; Recombinant Polypeptide ; Conformational Quality ; Recombinant Protein Yield
Published in: Microbial cell factories, Vol. 10 (October 2011) , art. 79, ISSN 1475-2859

PMID: 21992454
DOI: 10.1186/1475-2859-10-79

7 p, 370.3 KB

The record appears in these collections:
Articles > Research articles
Articles > Published articles

 Record created 2018-01-25, last modified 2019-03-20

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