New Vectors for Urea-Inducible Recombinant Protein Production

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 (BamAENm). 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.

Publication DOI: https://doi.org/10.1016/j.nbt.2022.10.003
Divisions: College of Health & Life Sciences > School of Biosciences
Additional Information: © 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/). Funding statement: 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).
Uncontrolled Keywords: Escherichia coli,Recombinant protein production,Biopharmaceuticals,Transcription regulation,ureR,Urea
Publication ISSN: 1876-4347
Last Modified: 28 Feb 2024 08:22
Date Deposited: 26 Oct 2022 15:16
Full Text Link:
Related URLs: https://www.sci ... 871678422000577 (Publisher URL)
PURE Output Type: Article
Published Date: 2022-12-25
Published Online Date: 2022-10-20
Accepted Date: 2022-10-19
Authors: Hothersall, Joanne
Osgerby, Alexander
Godfrey, Rita E
Overton, Tim W
Busby, Stephen J W
Browning, Douglas F (ORCID Profile 0000-0003-4672-3514)

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