Analysis of bioavailable toluene by using recombinant luminescent bacterial biosensors with different promoters.

In this study, we constructed recombinant luminescent Escherichia coli with T7, T3, and SP6 promoters inserted between tol and lux genes as toluene biosensors and evaluated their sensitivity, selectivity, and specificity for measuring bioavailable toluene in groundwater and river water. The luminescence intensity of each biosensor depended on temperature, incubation time, ionic strength, and concentrations of toluene and coexisting organic compounds. Toluene induced the highest luminescence intensity in recombinant lux-expressing E. coli with the T7 promoter [T7-lux-E. coli, limit of detection (LOD) = 0.05 µM], followed by that in E. coli with the T3 promoter (T3-lux-E. coli, LOD = 0.2 µM) and SP6 promoter (SP6-lux-E. coli, LOD = 0.5 µM). Luminescence may have been synergistically or antagonistically affected by coexisting organic compounds other than toluene; nevertheless, low concentrations of benzoate and toluene analogs had no such effect. In reproducibility experiments, the biosensors had low relative standard deviation (4.3-5.8%). SP6-lux-E. coli demonstrated high adaptability to environmental interference. T7-lux-E. coli biosensor-with low LOD, wide measurement range (0.05-500 µM), and acceptable deviation (- 14.3 to 9.1%)-is an efficient toluene biosensor. This is the first study evaluating recombinant lux E. coli with different promoters for their potential application in toluene measurement in actual water bodies.

Structure of the bacterial community in a biofilter during dimethyl sulfide (DMS) removal processes.

We report here both the successful treatment of DMS in a biofilter and the dynamic changes that occur in the composition of the bacterial community of the biofilter during this process. Denaturing gradient gel electrophoresis of eubacterial 16S rDNA samples taken from packing material at different DMS removal stages revealed 11 distinct bands. Phylogenetic analysis showed that the sequences of these bands were closest to sequences of species of the Proteobacteria, Firmicutes, and Actinobacteria. The specific occurrence of these bacterial species varied mainly with DMS load, but it was also affected by the addition of glucose and by ambient temperature. Based on the characteristics of the identified species, the system is conducive for such processes as sulfur oxidation, sulfate reduction, carbon oxidation, and fermentation. The strains identified in this study are potential candidates for purifying waste gas effluents containing DMS gas.