The construction and application of a lux-based nitrate biosensor.
A plasmid-borne transcriptional fusion between the Escherichia coli nitrate reductase (narG) promoter and the Photorhabdus luminescens lux 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 x 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 homologues 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.
Prest
A G
AG
Bass Technical Centre, Burton on Trent, UK.
Winson
M K
MK
Hammond
J R
JR
Stewart
G S
GS
D001483:Base Sequence; D015374:Biosensing Techniques; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D005504:Food Analysis; D005516:Food Microbiology; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000235:genetics; D009566:Nitrates / Q000032:analysis*; D009876:Operon; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D017931:DNA Primers; D009566:Nitrates; D008156:Luciferases; D009565:Nitrate Reductases; D050901:Nitrate Reductase