A secretory system for bacterial production of high-profile protein targets.

  • Alexander Kotzsch, Erik Vernet, +4 authors Michael Sundström
  • Published 2011 in
    Protein science : a publication of the Protein…

Abstract

Escherichia coli represents a robust, inexpensive expression host for the production of recombinant proteins. However, one major limitation is that certain protein classes do not express well in a biologically relevant form using standard expression approaches in the cytoplasm of E. coli. To improve the usefulness of the E. coli expression platform we have investigated combinations of promoters and selected N-terminal fusion tags for the extracellular expression of human target proteins. A comparative study was conducted on 24 target proteins fused to outer membrane protein A (OmpA), outer membrane protein F (OmpF) and osmotically inducible protein Y (OsmY). Based on the results of this initial study, we carried out an extended expression screen employing the OsmY fusion and multiple constructs of a more diverse set of human proteins. Using this high-throughput compatible system, we clearly demonstrate that secreted biomedically relevant human proteins can be efficiently retrieved and purified from the growth medium.

DOI: 10.1002/pro.593
050100201220132014201520162017
Citations per Year

150 Citations

Semantic Scholar estimates that this publication has 150 citations based on the available data.

See our FAQ for additional information.

Cite this paper

@article{Kotzsch2011ASS, title={A secretory system for bacterial production of high-profile protein targets.}, author={Alexander Kotzsch and Erik Vernet and Martin Hammarstr{\"{o}m and J. E. Berthelsen and J. Weigelt and Susanne Gr{\"a}slund and Michael Sundstr{\"{o}m}, journal={Protein science : a publication of the Protein Society}, year={2011}, volume={20 3}, pages={597-609} }