Inexpensive protein overexpression driven by the NarL transcription activator protein

Joanne Hothersall, Sandie Lai, Nan Zhang, Rita E Godfrey, Patcharawarin Ruanto, Sarah Bischoff, Colin Robinson, Tim W Overton, Stephen J W Busby, Douglas F Browning

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Pages (from-to)1614-1623
Number of pages10
JournalBiotechnology and Bioengineering
Issue number6
Early online date25 Feb 2022
Publication statusPublished - Jun 2022

Bibliographical note

© 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.


  • Escherichia coli
  • biopharmaceuticals
  • recombinant protein production
  • transcription regulation


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