New vectors for urea-inducible recombinant protein production

Joanne Hothersall; Alexander Osgerby; Rita Godfrey; Tim Overton; Steve Busby; Doug Browning

doi:10.1016/j.nbt.2022.10.003

New vectors for urea-inducible recombinant protein production

Joanne Hothersall, Alexander Osgerby, Rita Godfrey, Tim Overton, Steve Busby, Doug Browning

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Abstract

We have developed a novel urea-inducible recombinant protein production system by exploiting the Proteus mirabilis urease ureR-ureD promoter region and the ureR AraC-family transcriptional regulator. Experiments using the expression of β-galactosidase and green fluorescent protein (GFP) showed that promoter activity is tightly regulated and that varying the concentration of urea can give up to 100-fold induction. Production of proteins of biopharmaceutical interest has been demonstrated, including human growth hormone (hGH), a single chain antibody fragment (scFv) against interleukin-1β and a potential Neisserial vaccine candidate (BamA _ENm). Expression levels can be fine-tuned by temperature and different urea concentrations, and can be induced with readily available garden fertilisers and even urine. As urea is an inexpensive, stable inducer, a urea-induced expression system has the potential to considerably reduce the costs of large-scale recombinant protein production.

Original language	English
Pages (from-to)	89-96
Number of pages	8
Journal	New Biotechnology
Volume	72
Early online date	20 Oct 2022
DOIs	https://doi.org/10.1016/j.nbt.2022.10.003
Publication status	Published - 25 Dec 2022

Bibliographical note

Funding Information:
J.H. was generously funded by an Industrial Biotechnology Catalyst (Innovate UK, BBSRC, EPSRC) ( BB/M018261/1 ) to support the translation, development and commercialisation 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 . AO was funded by a BBSRC PhD studentship as part of the Midlands Integrative Biosciences Training Partnership ( MIBTP ).

Publisher Copyright:
© 2022 The Authors

Keywords

Biopharmaceuticals
Escherichia coli
Recombinant protein production
Transcription regulation
UreR
Urea

ASJC Scopus subject areas

Bioengineering
Molecular Biology
Biotechnology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.nbt.2022.10.003Licence: Creative Commons: Attribution (CC BY)

HothersallJ2022NewFinal published version, 3.91 MBLicence: Creative Commons: Attribution (CC BY)

BBSRC IAA: Making Recombinant Protein Production as â€œcheap as chipsâ€.
Busby, S. & Browning, D.
Biotechnology & Biological Sciences Research Council
1/12/19 → 31/08/20
Project: Research Councils
Understanding and exploiting regulation in pathogenic enteroaggregative Escherichia coli
Busby, S., Browning, D. & Henderson, I.
Biotechnology & Biological Sciences Research Council
1/01/19 → 31/12/21
Project: Research Councils
A new generation of E. coli expression hosts and tools for recombinant protein production
Dafforn, T., Browning, D. & Busby, S.
Biotechnology & Biological Sciences Research Council
1/04/15 → 3/10/19
Project: Research Councils

Cite this

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abstract = "We have developed a novel urea-inducible recombinant protein production system by exploiting the Proteus mirabilis urease ureR-ureD promoter region and the ureR AraC-family transcriptional regulator. Experiments using the expression of β-galactosidase and green fluorescent protein (GFP) showed that promoter activity is tightly regulated and that varying the concentration of urea can give up to 100-fold induction. Production of proteins of biopharmaceutical interest has been demonstrated, including human growth hormone (hGH), a single chain antibody fragment (scFv) against interleukin-1β and a potential Neisserial vaccine candidate (BamA ENm). Expression levels can be fine-tuned by temperature and different urea concentrations, and can be induced with readily available garden fertilisers and even urine. As urea is an inexpensive, stable inducer, a urea-induced expression system has the potential to considerably reduce the costs of large-scale recombinant protein production.",

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New vectors for urea-inducible recombinant protein production

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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Projects

BBSRC IAA: Making Recombinant Protein Production as â€œcheap as chipsâ€.

Understanding and exploiting regulation in pathogenic enteroaggregative Escherichia coli

A new generation of E. coli expression hosts and tools for recombinant protein production

Cite this

BBSRC IAA: Making Recombinant Protein Production as â€œcheap as chipsâ€.