Kinetic analysis of a tod-lux bacterial reporter for toluene degradation and trichloroethylene cometabolism

• Published in: Biotechnology and bioengineering Vol. 69; no. 3; pp. 256 - 265
• Main Authors: Kelly, C. J., Bienkowski, P. R., Sayler, G. S.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 05-08-2000; Wiley
• Subjects: Bacteria; Batch cell culture; Biodegradation; Biodegradation of pollutants; Biodegradation, Environmental - drug effects; Biological and medical sciences; bioluminescence; Biomass; Bioreactors; bioreporter; Biotechnology; Cell Division - drug effects; Dose-Response Relationship, Drug; Environment and pollution; Enzymes; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; Genes, Bacterial - genetics; Genes, Reporter; Industrial applications and implications. Economical aspects; Kinetics; Luciferases - genetics; Luciferases - metabolism; Luminescence; lux; Metabolism; Models, Biological; Oxygenases - genetics; Oxygenases - metabolism; Promoter Regions, Genetic; Pseudomonas putida; Pseudomonas putida - drug effects; Pseudomonas putida - enzymology; Pseudomonas putida - genetics; Pseudomonas putida - metabolism; Toluene; Toluene - metabolism; Toluene - pharmacology; trichloroethylene; trichloroethylene (TCE); Trichloroethylene - metabolism; Water Pollutants - metabolism; Pseudomonadales; Biodegradation; Toluene dioxygenase; Bioluminescence; Enzyme; Luciferase; Toluene; Cometabolism; Continuous process; Kinetic model; Pollutant; Reporter gene; Batchwise; Concentration effect; Microorganism culture; Bacteria; Pseudomonadaceae; Oxidoreductases; Bioremediation; Pseudomonas putida; Ethylene(trichloro)
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/1097-0290(20000805)69:3<256::AID-BIT3>3.0.CO;2-2

Kinetics of toluene and trichloroethylene (TCE) degradation and bioluminescence from the bioreporter Pseudomonas putida B2 and TVA8 were investigated utilizing batch and continuous culture, respectively. Degradation was modeled using a Michaelis–Menten expression for the competition of two substrates for a single enzyme system, and bioluminescence was modeled assuming a luciferase enzyme saturational dependence on toluene as the inducer and growth substrate. During the batch experiments, bioluminescence increased at approximately 90 namp/min for initial toluene concentrations of 10 to 50 mg/L, but more slowly at higher toluene concentrations, suggesting maximum promoter induction at below 10 mg/L and toxic effects above 50 mg/L toluene. TCE degradation did not occur until toluene depletion, presumably due to competition between toluene and TCE for the toluene dioxygenase enzyme. During continuous culture, bioluminescence transiently increased, then gradually decreased in response to increasing step changes in toluene feed concentration. Bioluminescence in the CSTR appeared to be limited by growth substrate and/or inducer. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 69: 256–265, 2000.