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

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

A secretory system for bacterial production of high-profile protein targets. / Kotzsch, Alexander; Vernet, Erik; Hammarström, Martin; Berthelsen, Jens; Weigelt, Johan; Gräslund, Susanne; Sundström, Michael.

In: Protein Science, Vol. 20, No. 3, 2011, p. 597-609.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Kotzsch, A, Vernet, E, Hammarström, M, Berthelsen, J, Weigelt, J, Gräslund, S & Sundström, M 2011, 'A secretory system for bacterial production of high-profile protein targets', Protein Science, vol. 20, no. 3, pp. 597-609. https://doi.org/10.1002/pro.593

APA

Kotzsch, A., Vernet, E., Hammarström, M., Berthelsen, J., Weigelt, J., Gräslund, S., & Sundström, M. (2011). A secretory system for bacterial production of high-profile protein targets. Protein Science, 20(3), 597-609. https://doi.org/10.1002/pro.593

Vancouver

Kotzsch A, Vernet E, Hammarström M, Berthelsen J, Weigelt J, Gräslund S et al. A secretory system for bacterial production of high-profile protein targets. Protein Science. 2011;20(3):597-609. https://doi.org/10.1002/pro.593

Author

Kotzsch, Alexander ; Vernet, Erik ; Hammarström, Martin ; Berthelsen, Jens ; Weigelt, Johan ; Gräslund, Susanne ; Sundström, Michael. / A secretory system for bacterial production of high-profile protein targets. In: Protein Science. 2011 ; Vol. 20, No. 3. pp. 597-609.

Bibtex

@article{50c774f27a2b4f91855d88010dd9a151,
title = "A secretory system for bacterial production of high-profile protein targets",
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.",
keywords = "Bacterial Outer Membrane Proteins, Bacterial Secretion Systems, Escherichia coli, Escherichia coli Proteins, Humans, Mass Spectrometry, Periplasmic Binding Proteins, Porins, Promoter Regions, Genetic, Recombinant Fusion Proteins",
author = "Alexander Kotzsch and Erik Vernet and Martin Hammarstr{\"o}m and Jens Berthelsen and Johan Weigelt and Susanne Gr{\"a}slund and Michael Sundstr{\"o}m",
note = "Copyright {\textcopyright} 2011 The Protein Society.",
year = "2011",
doi = "10.1002/pro.593",
language = "English",
volume = "20",
pages = "597--609",
journal = "Protein Science",
issn = "0961-8368",
publisher = "Wiley-Blackwell",
number = "3",

}

RIS

TY - JOUR

T1 - A secretory system for bacterial production of high-profile protein targets

AU - Kotzsch, Alexander

AU - Vernet, Erik

AU - Hammarström, Martin

AU - Berthelsen, Jens

AU - Weigelt, Johan

AU - Gräslund, Susanne

AU - Sundström, Michael

N1 - Copyright © 2011 The Protein Society.

PY - 2011

Y1 - 2011

N2 - 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.

AB - 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.

KW - Bacterial Outer Membrane Proteins

KW - Bacterial Secretion Systems

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Humans

KW - Mass Spectrometry

KW - Periplasmic Binding Proteins

KW - Porins

KW - Promoter Regions, Genetic

KW - Recombinant Fusion Proteins

U2 - 10.1002/pro.593

DO - 10.1002/pro.593

M3 - Journal article

C2 - 21308845

VL - 20

SP - 597

EP - 609

JO - Protein Science

JF - Protein Science

SN - 0961-8368

IS - 3

ER -

ID: 40289591