A bioluminescent reporter strain utilizing the lower pathway promoter (Pm) of the xyl operon of Pseudomonas: optimization of a bioassay for m-toluate

Kong, In Chul; Suh, Hak-Soo; Yang, Zamin K.; Burlage, Robert S.

doi:10.1016/s1093-0191(03)00037-6

Cited by 17 publications

(21 citation statements)

References 8 publications

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“…The tested recombinant strain Pseudomonas putida mt-2 KG1206 (called KG1206)contains the intact TOL plasmid and P m - lux fusion plasmid, where P m is the promoter of the lower or meta operon (13 genes) of the TOL catabolic plasmid pWW0 and is responsible for producing bioluminescence in the presence of toluene analogs (indirect inducers; toluene, m -MBA, xylene isomers) as well as their metabolites (direct inducer; m -toluate, and benzoate) ( Figure 1 ) [ 8 , 9 ]. Intermediates of toluene analogs, such as benzoate and m -toluate, activate XylS regulatory protein, whereas toluene, m -MBA, and isomers of xylene activate XylR regulatory protein.…”

Section: Methodsmentioning

confidence: 99%

“…When combined with genes that are activated in the presence of specific chemicals, the lux gene sequence of V. fischeri acts as an effective bioreporter for environmental monitoring applications. Under appropriate conditions, a recombinant bioreporter gene can produce a signal that is directly correlated to the concentration of a specific pollutant [ 9 ]. Such biosensors can generate valuable information about the bioavailability of contaminants in the environment and can serve as useful alternatives to more expensive chemical methods for in situ monitoring or on-site analysis.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Binary Mixtures of Inducers (Toluene Analogs) and of Metals on Bioluminescence Induction of a Recombinant Bioreporter Strain

Kong

2014

Sensors

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This paper investigated the effects of binary mixtures of bioluminescence inducers (toluene, xylene isomers, m-toluate) and of metals (Cu, Cd, As(III), As(V), and Cr) on bioluminescence activity of recombinant (Pm-lux) strain KG1206. Different responses and sensitivities were observed depending on the types and concentrations of mixtures of inducers or metals. In the case of inducer mixtures, antagonistic and synergistic modes of action were observed, whereas metal mixtures showed all three modes of action. Antagonistic mode of action was most common for mixtures of indirect inducers, which showed bioluminescence ranging from 29% to 62% of theoretically expected effects (P(E)). On the other hand, synergistic mode of action was observed for mixtures of direct and indirect inducers, which showed bioluminescence between 141% and 243% of P(E).In the case of binary metal mixtures, bioluminescence activities were ranged from 62% to 75% and 113% to 164% of P(E) for antagonistic and synergistic modes of action, respectively (p-values 0.0001–0.038). Therefore, mixture effects could not be generalized since they were dependent on both the types and concentrations of chemicals, suggesting that biomonitoring may constitute a better strategy by investigating types and concentrations of mixture pollutants at contaminated sites.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effects of Binary Mixtures of Inducers (Toluene Analogs) and of Metals on Bioluminescence Induction of a Recombinant Bioreporter Strain

Kong

2014

Sensors

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“…For the second, the bioreporter strain Pseudomonas putida mt-2 KG1206 (called KG1206 hereafter), containing the intact TOL plasmid and a Pm-lux fusion plasmid, was used. This strain produces bioluminescence in the presence of toluene analogs and intermediates [53]. To determine the effects of the Co-NPs, an inducer chemical solution (final 1 mM o-chlorotoluene) was added to the mixture solution of culture and sample.…”

Section: Effects Of Co-nps On Bacterial Bioluminescence Activitymentioning

confidence: 99%

Comparative Effects of Particle Sizes of Cobalt Nanoparticles to Nine Biological Activities

Kong

Koh

et al. 2020

IJMS

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The differences in the toxicity of cobalt oxide nanoparticles (Co-NPs) of two different sizes were evaluated in the contexts of the activities of bacterial bioluminescence, xyl-lux gene, enzyme function and biosynthesis of β-galactosidase, bacterial gene mutation, algal growth, and plant seed germination and root/shoot growth. Each size of Co-NP exhibited a different level of toxicity (sensitivity) in each biological activity. No revertant mutagenic ratio (greater than 2.0) of Salmonella typhimurium TA 98 was observed under the test conditions in the case of gene-mutation experiments. Overall, the inhibitory effects on all five bacterial bioassays were greater than those on algal growth, seed germination, and root growth. However, in all cases, the small Co-NPs showed statistically greater (total average about two times) toxicity than the large Co-NPs, except in shoot growth, which showed no observable inhibition. These findings demonstrate that particle size may be an important physical factor determining the fate of Co-NPs in the environment. Moreover, combinations of results based on various biological activities and physicochemical properties, rather than only a single activity and property, would better facilitate accurate assessment of NPs’ toxicity in ecosystems.

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“…Effects of coexisting carbon sources, intermediates, and toluene analogs on luminescence activity Figure 4A illustrates the effects of coexisting carbon sources at 100 μM on the luminescence activity of T7-lux-E. coli and SP6-lux-E. coli. The tested chemicals are considered potential inhibitors or activators (indirect or direct inducers) of xylS and xylR and may deviate signi cantly or be additive effect in relation to theoretically expected effects, calculated on the basis of individual chemicals [12,31,32]. The current results demonstrated that the coexistence of sodium lactate or glycerin with toluene induced higher luminescence activity than did toluene alone.…”

Section: Effects Of Culture Conditions On Luminescence Activitymentioning

confidence: 65%

Analysis of Bioavailable Toluene by Using Recombinant Luminescent Bacterial Biosensors with Different Promoters

Wang

Tsai

Kui

et al. 2020

Preprint

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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 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 activities 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.

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A bioluminescent reporter strain utilizing the lower pathway promoter (Pm) of the xyl operon of Pseudomonas: optimization of a bioassay for m-toluate

Cited by 17 publications

References 8 publications

Effects of Binary Mixtures of Inducers (Toluene Analogs) and of Metals on Bioluminescence Induction of a Recombinant Bioreporter Strain

Effects of Binary Mixtures of Inducers (Toluene Analogs) and of Metals on Bioluminescence Induction of a Recombinant Bioreporter Strain

Comparative Effects of Particle Sizes of Cobalt Nanoparticles to Nine Biological Activities

Analysis of Bioavailable Toluene by Using Recombinant Luminescent Bacterial Biosensors with Different Promoters

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