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Applied and Environmental Microbiology, August 1999, p. 3473-3482, Vol. 65, No. 8
Department of Soil, Water, and Environmental
Science, University of Arizona, Tucson, Arizona 85721
Received 26 February 1999/Accepted 26 May 1999
Bioluminescent reporter organisms have been successfully exploited
as analytical tools for in situ determination of bioavailable levels of
contaminants in static environmental samples. Continued characterization and development of such reporter systems is needed to
extend the application of these bioreporters to in situ monitoring of
degradation in dynamic environmental systems. In this study, the
naphthalene-degrading, lux bioreporter bacterium
Pseudomonas putida RB1353 was used to evaluate the relative
influences of cell growth stage, cell density, substrate concentration,
oxygen tension, and background carbon substrates on both the magnitude of the light response and the rate of salicylate disappearance. The
effect of these variables on the lag time required to obtain maximum
luminescence and degradation was also monitored. Strong correlations
were observed between the first three factors and both the magnitude
and induction time of luminescence and degradation rate. The maximum
luminescence response to nonspecific background carbon substrates (soil
extract broth or Luria broth) was 50% lower than that generated in
response to 1 mg of sodium salicylate liter
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Copyright © 1999, American Society for Microbiology. All rights reserved.
Factors Influencing Expression of
luxCDABE and nah Genes in Pseudomonas
putida RB1353(NAH7, pUTK9) in Dynamic Systems
1. Oxygen
tension was evaluated over the range of 0.5 to 40 mg liter
1, with parallel inhibition to luminescence and
degradation rate (20 mg of sodium salicylate liter
1)
observed at 1.5 mg liter
1 and below and no effect
observed above 5 mg liter
1. Oxygen tensions from 2 to 4 mg liter
1 influenced the magnitude of luminescence but
not the salicylate degradation rate. The results suggest that factors
causing parallel shifts in the magnitude of both luminescence and
degradation rate were influencing regulation of the nah
operon promoters. For factors that cause nonparallel shifts, other
regulatory mechanisms are explored. This study demonstrates that
lux reporter bacteria can be used to monitor both substrate
concentration and metabolic response in dynamic systems. However, each
lux reporter system and application will require
characterization and calibration.
*
Corresponding author. Mailing address: 429 Shantz
Building, Department of Soil, Water, and Environmental Science,
University of Arizona, P.O. Box 210038, Tucson, AZ 85721-0038. Phone:
(520) 621-7231. Fax: (520) 621-1647. E-mail:
rmaier{at}ag.arizona.edu.
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