Details
| Original language | English |
|---|---|
| Pages (from-to) | 247-258 |
| Number of pages | 12 |
| Journal | Applied Microbiology and Biotechnology |
| Volume | 105 |
| Early online date | 3 Dec 2020 |
| Publication status | Published - Jan 2021 |
Abstract
Abstract: Pseudomonads are among the most common bacteria in soils, limnic ecosystems, and human, animal, or plant host environments, including intensively studied species such as Pseudomonas aeruginosa, P. putida, or P. fluorescens. Various gene expression systems are established for some species, but there is still a need for a simple system that is suitable for a wide range of pseudomonads and that can be used for physiological applications, i.e., with a tuning capacity at lower expression levels. Here, we report the establishment of the anthranilate-dependent PantA promoter for tunable gene expression in pseudomonads. During studies on P. fluorescens, we constructed an anthranilate-inducible AntR/PantA-based expression system, named pUCP20-ANT, and used GFP as reporter to analyze gene expression. This system was compared with the rhamnose-inducible RhaSR/PrhaB-based expression system in an otherwise identical vector background. While the rhamnose-inducible system did not respond to lower inducer concentrations and always reached high levels over time when induced, expression levels of the pUCP20-ANT system could be adjusted to a range of distinct lower or higher levels by variation of anthranilate concentrations in the medium. Importantly, the anthranilate-inducible expression system worked also in strains of P. aeruginosa and P. putida and therefore will be most likely useful for physiological and biotechnological purposes in a wide range of pseudomonads. Key points: • We established an anthranilate-inducible gene expression system for pseudomonads. • This system permits tuning of gene expression in a wide range of pseudomonads. • It will be very useful for physiological and biotechnological applications.
Keywords
- Anthranilate, Gene regulation, Pseudomonas aeruginosa, Pseudomonas fluorescens, Recombinant expression
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Applied Microbiology and Biotechnology, Vol. 105, 01.2021, p. 247-258.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - A tunable anthranilate-inducible gene expression system for Pseudomonas species
AU - Hoffmann, Lena
AU - Sugue, Michael Frederick
AU - Brüser, Thomas
N1 - Funding Information: We thank Katrin Gunka for manifold suggestions and excellent technical assistance, and Jochen Meens and Nina Janze of the Institute of Microbiology, University of Veterinary Medicine Hannover (TiHo) for the support with flow cytometry. We are grateful to Michael Ringel for the support in early phases of this project. We thank Joyce E. Loper (Oregon State University) for kindly providing the strain P. fluorescens A506, and we also thank Susanne H?ussler (Institute of Molecular Bacteriology, TWINCORE, Hannover & Helmholtz Centre for Infection Research, Braunschweig) for donation of the vector pUCP20 and P. aeruginosa PAO1.
PY - 2021/1
Y1 - 2021/1
N2 - Abstract: Pseudomonads are among the most common bacteria in soils, limnic ecosystems, and human, animal, or plant host environments, including intensively studied species such as Pseudomonas aeruginosa, P. putida, or P. fluorescens. Various gene expression systems are established for some species, but there is still a need for a simple system that is suitable for a wide range of pseudomonads and that can be used for physiological applications, i.e., with a tuning capacity at lower expression levels. Here, we report the establishment of the anthranilate-dependent PantA promoter for tunable gene expression in pseudomonads. During studies on P. fluorescens, we constructed an anthranilate-inducible AntR/PantA-based expression system, named pUCP20-ANT, and used GFP as reporter to analyze gene expression. This system was compared with the rhamnose-inducible RhaSR/PrhaB-based expression system in an otherwise identical vector background. While the rhamnose-inducible system did not respond to lower inducer concentrations and always reached high levels over time when induced, expression levels of the pUCP20-ANT system could be adjusted to a range of distinct lower or higher levels by variation of anthranilate concentrations in the medium. Importantly, the anthranilate-inducible expression system worked also in strains of P. aeruginosa and P. putida and therefore will be most likely useful for physiological and biotechnological purposes in a wide range of pseudomonads. Key points: • We established an anthranilate-inducible gene expression system for pseudomonads. • This system permits tuning of gene expression in a wide range of pseudomonads. • It will be very useful for physiological and biotechnological applications.
AB - Abstract: Pseudomonads are among the most common bacteria in soils, limnic ecosystems, and human, animal, or plant host environments, including intensively studied species such as Pseudomonas aeruginosa, P. putida, or P. fluorescens. Various gene expression systems are established for some species, but there is still a need for a simple system that is suitable for a wide range of pseudomonads and that can be used for physiological applications, i.e., with a tuning capacity at lower expression levels. Here, we report the establishment of the anthranilate-dependent PantA promoter for tunable gene expression in pseudomonads. During studies on P. fluorescens, we constructed an anthranilate-inducible AntR/PantA-based expression system, named pUCP20-ANT, and used GFP as reporter to analyze gene expression. This system was compared with the rhamnose-inducible RhaSR/PrhaB-based expression system in an otherwise identical vector background. While the rhamnose-inducible system did not respond to lower inducer concentrations and always reached high levels over time when induced, expression levels of the pUCP20-ANT system could be adjusted to a range of distinct lower or higher levels by variation of anthranilate concentrations in the medium. Importantly, the anthranilate-inducible expression system worked also in strains of P. aeruginosa and P. putida and therefore will be most likely useful for physiological and biotechnological purposes in a wide range of pseudomonads. Key points: • We established an anthranilate-inducible gene expression system for pseudomonads. • This system permits tuning of gene expression in a wide range of pseudomonads. • It will be very useful for physiological and biotechnological applications.
KW - Anthranilate
KW - Gene regulation
KW - Pseudomonas aeruginosa
KW - Pseudomonas fluorescens
KW - Recombinant expression
KW - Gene expression
KW - Pseudomonas fluorescens
KW - Pseudomonas aeruginosa
KW - Recombinant expression
UR - http://www.scopus.com/inward/record.url?scp=85097172309&partnerID=8YFLogxK
U2 - 10.1007/s00253-020-11034-8
DO - 10.1007/s00253-020-11034-8
M3 - Article
C2 - 33270152
AN - SCOPUS:85097172309
VL - 105
SP - 247
EP - 258
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
ER -