Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Detection of Nitrate/Nitrite Bioavailability in Wastewater Using a luxCDABE-Based Klebsiella oxytoca Bioluminescent Bioreporter

  • Abd-El-Haleem, Desouky (Environmental Biotechnology Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), Mubarak City for Scientific Research and Technology Applications, New Burg-Elarab City) ;
  • Ripp, Steven (Center for Environmental Biotechnology, The University of Tennessee) ;
  • Zaki, Sahar (Environmental Biotechnology Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), Mubarak City for Scientific Research and Technology Applications, New Burg-Elarab City) ;
  • Sayler, Gary S. (Center for Environmental Biotechnology, The University of Tennessee)
  • Published : 2007.08.30
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Abstract

In 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.

Keywords

References

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