The Acinetobacter sp. chnB promoter together with its cognate positive regulator ChnR is an attractive new candidate for metabolic engineering applications in bacteria.
Over the last 10 years there has been an extremely fast development in the global characterization of bacteria at the genome, transcriptome, proteome and metabolome levels. To further explore and apply these complex data sets there is now a need for new biological tools that can be used to test or verify hypotheses generated on the basis of all the new information. Here, we report the integration of an expression cassette based on the Acinetobacter sp. chnB promoter and its cognate positive regulator chnR gene into a replicon derived from the broad-host-range plasmid RK2. Cyclohexanone was found to be the most efficient inducer of this system in Escherichia coli, using firefly luciferase as a reporter. To explore the potential of the system in another species, we show that the system can be used in combination with another similar expression cassette (Pm/xylS) to control the monomer composition of the industrially widely used exopolysaccharide alginate, produced by Pseudomonas fluorescens.
Steigedal
Magnus
M
Department of Biotechnology, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway.
Valla
Svein
S
D000150:Acinetobacter / Q000235:genetics* / Q000378:metabolism; D051777:AraC Transcription Factor; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D003512:Cyclohexanones / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D009876:Operon; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011551:Pseudomonas fluorescens / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012093:Replicon; D013045:Species Specificity; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*; D014157:Transcription Factors
D051777:AraC Transcription Factor; D001426:Bacterial Proteins; D003512:Cyclohexanones; D004268:DNA-Binding Proteins; D011135:Polysaccharides, Bacterial; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C078720:XylS protein, Pseudomonas putida; C400926:chnR protein, Acinetobacter; C064841:exopolysaccharide, Pseudomonas; C036468:cyclohexanone; D010105:Oxygenases; C019961:cyclohexanone oxygenase