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

doi:10.1186/1472-6750-7-32

BMC Biotechnology
Volume 7

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de Marco A
Deuerling E
Mogk A
Tomoyasu T
Bukau B

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Mogk A
Tomoyasu T
Bukau B
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Research article

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

Ario de Marco¹^,2 , Elke Deuerling³ , Axel Mogk³ , Toshifumi Tomoyasu⁴ and Bernd Bukau³

¹EMBL Heidelberg, Meyerhofstrasse 1, D-69117 Heidelberg, Germany

²IFOM-IEO Campus for Oncogenomics, via Adamello 16, I-20139, Milano, Italy

³ZMBH, Universität Heidelberg, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany

⁴Department of Microbiology and Molecular Genetics, Graduate School of Pharmaceutical Sciences, Chiba University, 1-33 Yayoicho, Inageku, Chiba 263-8522, Japan

author email corresponding author email

BMC Biotechnology 2007, 7:32doi:10.1186/1472-6750-7-32


Published:	12 June 2007

Abstract

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.

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.