New vectors for urea-inducible recombinant protein production

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...

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Veröffentlicht in:New biotechnology 2022-12, Vol.72, p.89-96
Hauptverfasser: Hothersall, Joanne, Osgerby, Alexander, Godfrey, Rita E., Overton, Tim W., Busby, Stephen J.W., Browning, Douglas F.
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Sprache:eng
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Zusammenfassung: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. •Urea-induced recombinant protein production using the P. mirabilis ureD promoter.•Production of high value biopharmaceuticals with the uti101 expression system.•Fine tuning of expression levels by inducer concentration and thermoregulation.•Inexpensive recombinant protein production induced by urea, fertilisers and urine.
ISSN:1871-6784
1876-4347
DOI:10.1016/j.nbt.2022.10.003