Monitoring of naphthalene catabolism by bioluminescence with nah-lux transcriptional fusions.
We have demonstrated the efficacy of a light-generating genetic construction in describing the induction of a nah operon for the catabolism of naphthalene. A fragment from plasmid NAH7, which contains the promoter for the upper pathway of degradation, was transcriptionally fused to the lux genes of Vibrio fischeri. A Pseudomonas strain containing this construction is inducible to high levels of light production in the presence of a suitable substrate and the nahR regulatory gene product. This system was used to examine catabolic activity in a unique manner under a variety of growth conditions. Induction of bioluminescence was demonstrated to coincide with naphthalene degradation in all cases through the use of mineralization assays. A significant delay in bioluminescence and biodegradation was observed when naphthalene was added to batch cultures that were growing exponentially. These results suggest that the metabolism of naphthalene by this Pseudomonas strain is optimal when the growth rate of the culture is slow and is greatly reduced during exponential growth.
Burlage
R S
RS
Center for Environmental Biotechnology, University of Tennessee, Knoxville 37996.
Sayler
G S
GS
Larimer
F
F
D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005809:Genes, Regulator; D007700:Kinetics; D008027:Light; D008163:Luminescent Measurements; D009281:Naphthalenes / Q000378:metabolism*; D009876:Operon; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014733:Vibrio / Q000235:genetics* / Q000378:metabolism
D009281:Naphthalenes; C031721:naphthalene