Inexpensive protein overexpression driven by the NarL transcription activator protein

Abstract

Most Escherichia coli overexpression vectors used for recombinant protein production (RPP) depend on organic inducers, for example, sugars or simple conjugates. However, these can be expensive and, sometimes, chemically unstable. To simplify this and to cut the cost of RPP, we have developed vectors controlled by the Escherichia coli nitrate-responsive NarL transcription activator protein, which use nitrate, a cheap, stable, and abundant inorganic ion, to induce high-level controlled RPP. We show that target proteins, such as green fluorescent protein, human growth hormone, and single-chain variable region antibody fragments can be expressed to high levels using our promoter systems. As nitrate levels are high in many commercial fertilizers, we demonstrate that controlled RPP can be achieved using readily available and inexpensive garden products.

Publication DOI: https://doi.org/10.1002/bit.28071
Divisions: College of Health & Life Sciences > School of Biosciences
Additional Information: © 2022 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Grant numbers/ Funding: Industrial Biotechnology Catalyst Grant (Innovate UK, BBSRC, EPSRC) (BB/M018261/1); BBSRC IAA Follow-on-Fund award (BBSRC IAA BB/S506709/1); BBSRC grant BB/R017689/1; BBSRC Midlands Integrative Biosciences Training Programme.
Uncontrolled Keywords: Escherichia coli,biopharmaceuticals,recombinant protein production,transcription regulation,Biotechnology,Bioengineering,Applied Microbiology and Biotechnology
Publication ISSN: 1097-0290
Last Modified: 04 Mar 2024 08:42
Date Deposited: 28 Feb 2022 17:54
Full Text Link:
Related URLs: https://onlinel ... .1002/bit.28071 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2022-06
Published Online Date: 2022-02-25
Accepted Date: 2022-02-20
Authors: Hothersall, Joanne
Lai, Sandie
Zhang, Nan
Godfrey, Rita E
Ruanto, Patcharawarin
Bischoff, Sarah
Robinson, Colin
Overton, Tim W
Busby, Stephen J W
Browning, Douglas F (ORCID Profile 0000-0003-4672-3514)

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