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.
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Posted 07 Jan, 2021
On 06 Jan, 2021
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Analysis of Bioavailable Toluene by Using Recombinant Luminescent Bacterial Biosensors with Different Promoters
Posted 07 Jan, 2021
On 06 Jan, 2021
On 27 Dec, 2020
On 25 Dec, 2020
On 25 Dec, 2020
On 25 Dec, 2020
Received 19 Dec, 2020
On 17 Dec, 2020
On 16 Dec, 2020
Invitations sent on 16 Dec, 2020
On 16 Dec, 2020
On 16 Dec, 2020
On 15 Nov, 2020
Received 14 Nov, 2020
On 11 Nov, 2020
On 06 Nov, 2020
Received 06 Nov, 2020
Invitations sent on 03 Nov, 2020
On 01 Nov, 2020
On 01 Nov, 2020
On 01 Nov, 2020
On 24 Oct, 2020
Received 05 Oct, 2020
On 02 Oct, 2020
Invitations sent on 29 Sep, 2020
On 28 Sep, 2020
On 27 Sep, 2020
On 27 Sep, 2020
On 24 Aug, 2020
Received 15 Aug, 2020
On 08 Aug, 2020
On 04 Aug, 2020
Invitations sent on 04 Aug, 2020
On 03 Aug, 2020
On 03 Aug, 2020
On 17 Jul, 2020
Received 26 Jun, 2020
On 12 Jun, 2020
Invitations sent on 02 May, 2020
On 28 Apr, 2020
On 27 Apr, 2020
On 27 Apr, 2020
On 25 Apr, 2020
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.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5