Journal of Microbiology and Biotechnology
The Korean Society for Microbiology and Biotechnology publishes the Journal of Microbiology and Biotechnology.
Journal of Microbiology and Biotechnology
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2007 ; 17(8): 1254~1261

AuthorGary S. Sayler, Steven Ripp, Abd-El-Haleem, Desoukky, Sahar Zaki
AffiliationCenter for Environmental Biotechnology, The University of Tennessee, Knoxville, TN 37996, U.S.A.
TitleDetection of Nitrate/Nitrite Bioavailability in Wastewater Using a luxCDABEBased Klebsiella oxytoca Bioluminescent Bioreporter
PublicationInfo J. Microbiol. Biotechnol.2007 17(8): 1254~1261
AbstractIn the present study, we have constructed a bioluminescent bioreporter for the assessment of nitrate/nitrite bioavailability in wastewater. Specifically, an approximately 500-bp DNA fragment containing a nitrate/nitrite-activated nasR-like promoter (regulating expression of genes encoding nitrite reductase in the genus Klebsiella) was fused upstream of the Vibrio fischeri luxCDABE gene cassette in a modified mini-Tn5 vector. Characterization of this strain, designated W6-1, yielded dose-dependent increased bioluminescence coincident with increased nitrate, nitrite, and ammonium added to the growth medium from 1 to 11 ppm. Bioluminescence in response to nitrogen species addition was light dependent up to 10, 7, and 8 ppm with nitrate, nitrite, and ammonium, respectively. This response was linear in the range from 1 to 8 ppm for nitrate ($R^2=0.98$), 1 to 6 ppm for nitrite ($R^2=0.99$), and 1 to 7 ppm for ammonium ($R^2=0.99$). A significant bioluminescent response was also recorded when strain W6-1 was incubated with slurries from aged, nitrate/nitrite contaminated wastewater. Thus, bioreporter strain W6-1 can be used to elucidate factors that constrain the use of nitrate/nitrite in wastewaters.
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KeywordsKlebsiella oxytoca, bioluminescence, bioreporter, lux, nasR, nitrate, ammonium, bioavailability



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