Bioluminescent bioreporter Pseudomonas putida TVA8 as a detector of water pollution. Operational conditions and selectivity of free cells sensor

• Published in: Ecological indicators Vol. 11; no. 3; pp. 882 - 887
• Main Authors: Kuncova, Gabriela, Pazlarova, Jarmila, Hlavata, Alena, Ripp, Steven, Sayler, Gary S.
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01.05.2011
• Subjects: benzene; Bioluminescence; Bioreporter; biosensors; BTEX; BTEX (benzene, toluene, ethylbenzene, xylene); cell growth; ethanol; groundwater; indicator species; lux; methanol; pollutants; Pseudomonas putida; Pseudomonas putida TVA8; temperature; toluene; wastewater; water pollution; Whole-cell biosensor; xylene; Pseudomonas putida TVA8; Bioluminescence; BTEX; Bioreporter; Whole-cell biosensor; lux
• ISSN: 1470-160X; 1872-7034
• DOI: 10.1016/j.ecolind.2010.12.001

Conditions influencing bioluminescence output from Pseudomonas putida TVA8 harboring chromosomal tod-luxCDABE fusion were followed. In complex media, cell growth was not influenced by the presence of toluene at 53mg/L. Bioluminescence induction was tested in minimal medium. At 15°C the highest bioluminescence induced with toluene (1.325mg/L) was reached after 6h. At 25°C the bioluminescence maximum was approximately 20% lower but this was reached after 3.5h, and at temperatures of 7°C, 28°C, 30°C and 34°C, bioluminescence peaked at ≤60% of the maximum. Time courses of bioluminescence were dependent on cell concentrations. The heights of bioluminescence maxima were proportional to toluene concentration in the range 0–26mg/L. Twenty-three organic pollutants (103× diluted saturated solutions) were tested as bioluminescent inducers. The bioluminescence maximum decreased in the order: ethylbenzene>toluene>phenol>benzene>4-ethyltoluene>4-fluorotoluene>cumene>isobutylbenzene>styrene>trichloroethylene>o-, p-xylene>cresol>m-xylene>2-methylnaphthalene>benzylchloride>naphthalene>salicylic acid>hexachlorobenzene>2-chloronaphthalene>biphenyl>2-bromonaphthalene>1,3,5-triethylbenzene. Bioluminescence was also induced with ethanol and methanol and the presence of these alcohols in concentrations of ≤1% increased bioluminescence of toluene. The induction of bioluminescence from samples of wastewater and groundwater contaminated with BTEX (benzene, toluene, ethylbenzene and xylene) from 0.5 to 120mg/L was demonstrated.