Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli

Kurzanzeige
dc.contributor.authorMarco, Ario dedeu
dc.contributor.authorDeuerling, Elke
dc.contributor.authorMogk, Axeldeu
dc.contributor.authorTomoyasu, Toshifumideu
dc.contributor.authorBukau, Bernddeu
dc.date.accessioned2011-03-24T17:44:27Zdeu
dc.date.available2011-03-24T17:44:27Zdeu
dc.date.issued2007deu
dc.description.abstractBackground: The overproduction of recombinant proteins in host cells often leads to their misfolding and aggregation. Previous attempts to increase the solubility of recombinant proteins by co-overproduction of individual chaperones were only partially successful. We now assessed the effects of combined overproduction of the functionally cooperating chaperone network of the E. coli cytosol on the solubility of recombinant proteins.

Results: A two-step procedure was found to show the strongest enhancement of solubility. In a first step, the four chaperone systems GroEL/GroES, DnaK/DnaJ/GrpE, ClpB and the small HSPs IbpA/IbpB, were coordinately co-overproduced with recombinant proteins to optimize de novo folding. In a second step, protein biosynthesis was inhibited to permit chaperone mediated refolding of misfolded and aggregated proteins in vivo. This novel strategy increased the solubility of 70% of 64 different heterologous proteins tested up to 42-fold.

Conclusion: The engineered E. coli strains and the two-step procedure presented here led to a remarkable increase in the solubility of a various recombinant proteins and should be applicable to a wide range of target proteins produced in biotechnology.		eng
dc.description.versionpublished
dc.format.mimetypeapplication/pdfdeu
dc.identifier.citationFirst publ. in: BMC Biotechnology (2007), 7:32deu
dc.identifier.doi10.1186/1472-6750-7-32
dc.identifier.pmid17565681
dc.identifier.ppn272608459deu
dc.identifier.urihttp://kops.uni-konstanz.de/handle/123456789/8532
dc.language.isoengdeu
dc.legacy.dateIssued2007deu
dc.rightsAttribution-NonCommercial-NoDerivs 2.0 Generic
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/2.0/
dc.subject.ddc570deu
dc.titleChaperone-based procedure to increase yields of soluble recombinant proteins produced in E. colieng
dc.typeJOURNAL_ARTICLEdeu
dspace.entity.typePublication
kops.citation.bibtex
@article{Marco2007Chape-8532,
  year={2007},
  doi={10.1186/1472-6750-7-32},
  title={Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli},
  number={32},
  volume={7},
  issn={1472-6750},
  journal={BMC Biotechnology},
  author={Marco, Ario de and Deuerling, Elke and Mogk, Axel and Tomoyasu, Toshifumi and Bukau, Bernd}
}
kops.citation.iso690MARCO, Ario de, Elke DEUERLING, Axel MOGK, Toshifumi TOMOYASU, Bernd BUKAU, 2007. Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli. In: BMC Biotechnology. 2007, 7(32), pp. 32. ISSN 1472-6750. eISSN 1472-6750. Available under: doi: 10.1186/1472-6750-7-32deu
kops.citation.iso690MARCO, Ario de, Elke DEUERLING, Axel MOGK, Toshifumi TOMOYASU, Bernd BUKAU, 2007. Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli. In: BMC Biotechnology. 2007, 7(32), pp. 32. ISSN 1472-6750. eISSN 1472-6750. Available under: doi: 10.1186/1472-6750-7-32eng
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    <dcterms:abstract xml:lang="eng">Background: The overproduction of recombinant proteins in host cells often leads to their misfolding and aggregation. Previous attempts to increase the solubility of recombinant proteins by co-overproduction of individual chaperones were only partially successful. We now assessed the effects of combined overproduction of the functionally cooperating chaperone network of the E. coli cytosol on the solubility of recombinant proteins.&lt;br /&gt;&lt;br /&gt;Results: A two-step procedure was found to show the strongest enhancement of solubility. In a first step, the four chaperone systems GroEL/GroES, DnaK/DnaJ/GrpE, ClpB and the small HSPs IbpA/IbpB, were coordinately co-overproduced with recombinant proteins to optimize de novo folding. In a second step, protein biosynthesis was inhibited to permit chaperone mediated refolding of misfolded and aggregated proteins in vivo. This novel strategy increased the solubility of 70% of 64 different heterologous proteins tested up to 42-fold.&lt;br /&gt;&lt;br /&gt;Conclusion: The engineered E. coli strains and the two-step procedure presented here led to a remarkable increase in the solubility of a various recombinant proteins and should be applicable to a wide range of target proteins produced in biotechnology.</dcterms:abstract>
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kops.sourcefieldBMC Biotechnology. 2007, <b>7</b>(32), pp. 32. ISSN 1472-6750. eISSN 1472-6750. Available under: doi: 10.1186/1472-6750-7-32deu
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kops.sourcefield.plainBMC Biotechnology. 2007, 7(32), pp. 32. ISSN 1472-6750. eISSN 1472-6750. Available under: doi: 10.1186/1472-6750-7-32eng
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