construction and application of a lux-based nitrate biosensor

• Published in: Letters in applied microbiology Vol. 24; no. 5; pp. 355 - 360
• Main Authors: Prest, A.G, Winson, M.K, Hammond, J.R.M, Stewart, G.S.A.B
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.05.1997; Blackwell Science
• Subjects: Base Sequence; Beers; Biological and medical sciences; Biosensing Techniques; Biosensors; Biotechnology; brewing; Cloning, Molecular; DNA Primers - genetics; Escherichia coli; Escherichia coli - genetics; Fermentation; Fermented food industries; Food Analysis; Food industries; Food Microbiology; Fundamental and applied biological sciences. Psychology; Genes, Reporter; luciferase; Luciferases - genetics; Luminescent Measurements; Methods. Procedures. Technologies; Nitrate Reductase; Nitrate Reductases - genetics; Nitrates - analysis; Operon; Photorhabdus luminescens; Polymerase Chain Reaction; Promoter Regions, Genetic; Various methods and equipments; Brewery; Bioluminescence; Flavobacteriaceae; Enzyme; Escherichia coli; Nitrates; Gene expression; Flavobacterium proteus; Plasmid; Nitrate reductase; Biosensor; Pathogenic; Bacteria; Oxidoreductases; Detection; Biological contamination; Hybrid gene; Enterobacteriaceae
• ISSN: 0266-8254; 1472-765X
• DOI: 10.1046/j.1472-765X.1997.00064.x

A plasmid‐borne transcriptional fusion between the Escherichia coli nitrate reductase (narG) promoter and the Photorhabdus luminescenslux operon provides E. coli with a highly bioluminescent phenotype in the presence of nitrate. This E. coli biosensor can detect nitrate to a level of 5×10−5 mol l−1 (0·3 ppm), levels relevant to those levels encountered in brewing water. Since induction of the narG promoter requires NarL, the plasmid‐based sensor can also be used to interrogate enteric bacteria for the presence of functional xrhomologues of this E. coli regulatory protein. Obesumbacterium proteus, an important bacterial brewery contaminant, failed to provide nitrate‐dependent bioluminescence demonstrating divergence in this organism from E. coli in the mechanism of nitrate reductase regulation.