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Analysis of Bioavailable Toluene by Using Recombinant Luminescent Bacterial Biosensors with Different Promoters
Ying-Chien Chung
China University of Science and Technology
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0425-5444
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Journal of Biological Engineering.
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.
Keywords
Biosensor, Groundwater, Promoter, Toluene
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CURRENT STATUS: Published
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Posted 07 Jan, 2021
Published
On 06 Jan, 2021
No community comments so far
Editorial decision: Accept
On 27 Dec, 2020
Editor assigned
On 25 Dec, 2020
Submission checks complete
On 25 Dec, 2020
Editor invited
On 25 Dec, 2020
Review #1 received
Received 19 Dec, 2020
Reviewer #1 agreed
On 17 Dec, 2020
Editor assigned
On 16 Dec, 2020
Reviewers invited
Invitations sent on 16 Dec, 2020
Submission checks complete
On 16 Dec, 2020
Editor invited
On 16 Dec, 2020
No comments provided
Editorial decision: Major revision
On 15 Nov, 2020
Review #2 received
Received 14 Nov, 2020
Reviewer #2 agreed
On 11 Nov, 2020
Reviewer #1 agreed
On 06 Nov, 2020
Review #1 received
Received 06 Nov, 2020
Reviewers invited
Invitations sent on 03 Nov, 2020
Editor assigned
On 01 Nov, 2020
Submission checks complete
On 01 Nov, 2020
Editor invited
On 01 Nov, 2020
No comments provided
Editorial decision: Major revision
On 24 Oct, 2020
Review #1 received
Received 05 Oct, 2020
Reviewer #1 agreed
On 02 Oct, 2020
Reviewers invited
Invitations sent on 29 Sep, 2020
Editor assigned
On 28 Sep, 2020
Submission checks complete
On 27 Sep, 2020
Editor invited
On 27 Sep, 2020
No comments provided
Editorial decision: Minor revision
On 24 Aug, 2020
Review #1 received
Received 15 Aug, 2020
Reviewer #1 agreed
On 08 Aug, 2020
Editor assigned
On 04 Aug, 2020
Reviewers invited
Invitations sent on 04 Aug, 2020
Submission checks complete
On 03 Aug, 2020
Editor invited
On 03 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 17 Jul, 2020
Review #1 received
Received 26 Jun, 2020
Reviewer #1 agreed
On 12 Jun, 2020
Reviewers invited
Invitations sent on 02 May, 2020
Editor assigned
On 28 Apr, 2020
Submission checks complete
On 27 Apr, 2020
Editor invited
On 27 Apr, 2020
First submitted
On 25 Apr, 2020
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This preprint is under consideration at Journal of Biological Engineering. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Analysis of Bioavailable Toluene by Using Recombinant Luminescent Bacterial Biosensors with Different Promoters
Ying-Chien Chung
China University of Science and Technology
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0425-5444
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 5
Posted 07 Jan, 2021
Published
On 06 Jan, 2021
No community comments so far
Editorial decision: Accept
On 27 Dec, 2020
Editor assigned
On 25 Dec, 2020
Submission checks complete
On 25 Dec, 2020
Editor invited
On 25 Dec, 2020
Review #1 received
Received 19 Dec, 2020
Reviewer #1 agreed
On 17 Dec, 2020
Editor assigned
On 16 Dec, 2020
Reviewers invited
Invitations sent on 16 Dec, 2020
Submission checks complete
On 16 Dec, 2020
Editor invited
On 16 Dec, 2020
No comments provided
Editorial decision: Major revision
On 15 Nov, 2020
Review #2 received
Received 14 Nov, 2020
Reviewer #2 agreed
On 11 Nov, 2020
Reviewer #1 agreed
On 06 Nov, 2020
Review #1 received
Received 06 Nov, 2020
Reviewers invited
Invitations sent on 03 Nov, 2020
Editor assigned
On 01 Nov, 2020
Submission checks complete
On 01 Nov, 2020
Editor invited
On 01 Nov, 2020
No comments provided
Editorial decision: Major revision
On 24 Oct, 2020
Review #1 received
Received 05 Oct, 2020
Reviewer #1 agreed
On 02 Oct, 2020
Reviewers invited
Invitations sent on 29 Sep, 2020
Editor assigned
On 28 Sep, 2020
Submission checks complete
On 27 Sep, 2020
Editor invited
On 27 Sep, 2020
No comments provided
Editorial decision: Minor revision
On 24 Aug, 2020
Review #1 received
Received 15 Aug, 2020
Reviewer #1 agreed
On 08 Aug, 2020
Editor assigned
On 04 Aug, 2020
Reviewers invited
Invitations sent on 04 Aug, 2020
Submission checks complete
On 03 Aug, 2020
Editor invited
On 03 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 17 Jul, 2020
Review #1 received
Received 26 Jun, 2020
Reviewer #1 agreed
On 12 Jun, 2020
Reviewers invited
Invitations sent on 02 May, 2020
Editor assigned
On 28 Apr, 2020
Submission checks complete
On 27 Apr, 2020
Editor invited
On 27 Apr, 2020
First submitted
On 25 Apr, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Journal of Biological Engineering.
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