New Vectors for Urea-Inducible Recombinant Protein Production

Joanne Hothersall; Alexander Osgerby; Rita E Godfrey; Tim W Overton; Stephen J W Busby; Douglas F Browning

doi:10.1016/j.nbt.2022.10.003

New Vectors for Urea-Inducible Recombinant Protein Production

Joanne Hothersall^*, Alexander Osgerby, Rita E Godfrey, Tim W Overton, Stephen J W Busby, Douglas F Browning^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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.

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

© 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).

Keywords

Escherichia coli
Recombinant protein production
Biopharmaceuticals
Transcription regulation
ureR
Urea

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10.1016/j.nbt.2022.10.003Licence: CC BY 4.0

New vectors for urea-inducible recombinant protein production
© 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/).
Final published version, 3.91 MBLicence: CC BY 4.0

Cite this

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title = "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.",

keywords = "Escherichia coli, Recombinant protein production, Biopharmaceuticals, Transcription regulation, ureR, Urea",

author = "Joanne Hothersall and Alexander Osgerby and Godfrey, {Rita E} and Overton, {Tim W} and Busby, {Stephen J W} and Browning, {Douglas F}",

note = "{\textcopyright} 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).",

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AU - Busby, Stephen J W

AU - Browning, Douglas F

N1 - © 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).

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N2 - 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.

AB - 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.

KW - Escherichia coli

KW - Recombinant protein production

KW - Biopharmaceuticals

KW - Transcription regulation

KW - ureR

KW - Urea

UR - https://www.sciencedirect.com/science/article/pii/S1871678422000577

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VL - 72

SP - 89

EP - 96

JO - New Biotechnology

JF - New Biotechnology

ER -

New Vectors for Urea-Inducible Recombinant Protein Production

Abstract

Bibliographical note

Keywords

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