Conditions required for the stimulation of bioluminescence activity of the genetically engineered bacteria, P. putida mt-2 KG1206, preserved by deep-freezing.
Herein the conditions required for the stimulation of bioluminescence activity in a genetically engineered strain of Pseudomonas putida mt-2 KG1206, containing the intact TOL plasmid and a constructed plasmid with the P(m)-lux gene, are reported upon. Both sodium lactate (SL) and potassium nitrate (KNO(3)) were able to stimulate the bioluminescence activity, but a greater increase was observed with nitrogen amendment. This selected stimulant was then tested on reconstituted cells that had been preserved by deep-freezing and mixed with pure inducer solution or groundwater samples. The stimulation of bioluminescence activities for deep-frozen strain was in the range of 101-238% of the control. The effect of KNO(3) was found to be dependent on the type of inducers used and the cell conditions. In general, high bioluminescence activity was observed with groundwater samples, contaminated with high inducer compounds. However, no significant correlation was observed between the bioluminescence intensity and the total inducer concentration in the environmental samples contaminated with complex mixtures with inducers. These results should be useful when other recombinant bioluminescence strains are to be used for environmental monitoring. Overall, the results of this study demonstrate the stimulant conditions for the bioluminescence activity of genetically engineered bacteria, and suggest the potential for preliminary application of this deep-frozen engineered strain in a field-ready bioassay to conveniently detect or monitor a specific group of environmental contaminants.
Ko
K S
KS
Geo-Environmental Division, Korea Institute of Geoscience and Mineral Resources, South Korea.
Kong
In Chul
IC
D015925:Cryopreservation; D004784:Environmental Monitoring / Q000379:methods; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics*; D009566:Nitrates / Q000494:pharmacology; D030781:Organisms, Genetically Modified / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D017680:Potassium Compounds / Q000494:pharmacology; D016958:Pseudomonas putida / Q000187:drug effects / Q000235:genetics*; D019354:Sodium Lactate / Q000494:pharmacology
D008164:Luminescent Proteins; D009566:Nitrates; D017680:Potassium Compounds; C023844:potassium nitrate; D019354:Sodium Lactate