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A secretory system for bacterial production of high‐profile protein targets
Author(s) -
Kotzsch Alexander,
Vernet Erik,
Hammarström Martin,
Berthelsen Jens,
Weigelt Johan,
Gräslund Susanne,
Sundström Michael
Publication year - 2011
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.593
Subject(s) - escherichia coli , fusion protein , bacterial outer membrane , cytoplasm , biology , recombinant dna , membrane protein , target protein , extracellular , microbiology and biotechnology , biochemistry , gene , membrane
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.