DOI: 10.1002/bit.10865. Efficient production of a bioactive, multiple disulfide‐bonded protein using modified extracts of <i>Escherichia coli</i>

Efficient production of a bioactive, multiple disulfide‐bonded protein using modified extracts of Escherichia coli

10.1002/bit.10865

Crossref journal-article

Wiley

Biotechnology and Bioengineering (311)

Abstract

AbstractIn this report, we demonstrate that a complex mammalian protein containing multiple disulfide bonds is successfully expressed in an E.coli‐based cell‐free protein synthesis system. Initially, disulfide‐reducing activities in the cell extract prevented the formation of disulfide bonds. However, a simple pretreatment of the cell extract with iodoacetamide abolished the reducing activity. This extract was still active for protein synthesis even under oxidizing conditions. The use of a glutathione redox buffer coupled with the DsbC disulfide isomerase and pH optimization produced 40 μg/mL of active urokinase protease in a simple batch reaction. This result not only demonstrates efficient production of complex proteins, it also emphasizes the control and flexibility offered by the cell‐free approach. © 2003 Wiley Periodicals, Inc.

Bibliography

Kim, D., & Swartz, J. R. (2003). Efficient production of a bioactive, multiple disulfideâbonded protein using modified extracts of Escherichia coli. Biotechnology and Bioengineering, 85(2), 122â129. Portico.

Authors 2

Dong‐Myung Kim (first)
James R. Swartz (additional)

References 25 Referenced 129

Dates

Type	When
Created	21 years, 8 months ago (Dec. 30, 2003, 4 p.m.)
Deposited	1 year, 9 months ago (Nov. 17, 2023, 9:04 a.m.)
Indexed	1 month, 1 week ago (July 23, 2025, 8:03 a.m.)
Issued	21 years, 9 months ago (Nov. 25, 2003)
Published	21 years, 9 months ago (Nov. 25, 2003)
Published Online	21 years, 9 months ago (Nov. 25, 2003)
Published Print	21 years, 7 months ago (Jan. 20, 2004)

Funders 0

None

BibTeX

@article{Kim_2003, title={Efficient production of a bioactive, multiple disulfide‐bonded protein using modified extracts of Escherichia coli}, volume={85}, ISSN={1097-0290}, url={http://dx.doi.org/10.1002/bit.10865}, DOI={10.1002/bit.10865}, number={2}, journal={Biotechnology and Bioengineering}, publisher={Wiley}, author={Kim, Dong‐Myung and Swartz, James R.}, year={2003}, month=nov, pages={122–129} }

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