Inexpensive protein overexpression driven by the NarL transcription activator protein

Joanne Hothersall, Sandie Lai, Nan Zhang, Rita Godfrey, Patcharawarin Ruanto, Sarah Bischoff, Colin Robinson, Tim Overton, Steve Busby, Doug Browning

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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.
Original languageEnglish
Pages (from-to)1614-1623
Number of pages10
JournalBiotechnology and Bioengineering
Volume119
Issue number6
Early online date25 Feb 2022
DOIs
Publication statusPublished - 13 May 2022

Bibliographical note

Funding Information:
J. H. was supported by an Industrial Biotechnology Catalyst (Innovate UK, BBSRC, EPSRC) (BB/M018261/1) to promote the translation, development, and commercialization of innovative Industrial Biotechnology processes, and by a BBSRC IAA Follow‐on‐Fund award (BBSRC IAA BB/S506709/1). D. F. B was supported by BBSRC grants BB/M018261/1 and BB/R017689/1. S. L. was supported by a studentship from the BBSRC Midlands Integrative Biosciences Training Programme. This study was supported by 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.

Publisher Copyright:
© 2022 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC

Keywords

  • Biopharmaceuticals
  • Escherichia coli
  • Recombinant protein production
  • transcription regulation
  • recombinant protein production
  • biopharmaceuticals
  • Nitrates/metabolism
  • Operon
  • Humans
  • Transcription Factors/genetics
  • Escherichia coli/genetics
  • DNA-Binding Proteins/genetics
  • Base Sequence
  • Escherichia coli Proteins/genetics
  • Transcription, Genetic
  • Gene Expression Regulation, Bacterial

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