DOI: 10.1128/aem.64.3.1006-1012.1998. Development and Testing of a Bacterial Biosensor for Toluene-Based Environmental Contaminants

Development and Testing of a Bacterial Biosensor for Toluene-Based Environmental Contaminants

10.1128/aem.64.3.1006-1012.1998

Crossref journal-article

American Society for Microbiology

Applied and Environmental Microbiology (235)

Abstract

ABSTRACT A bacterial biosensor for benzene, toluene, and similar compounds has been constructed, characterized, and field tested on contaminated water and soil. The biosensor is based on a plasmid incorporating the transcriptional activator xylR from the TOL plasmid of Pseudomonas putida mt-2. The XylR protein binds a subset of toluene-like compounds and activates transcription at its promoter, P u . A reporter plasmid was constructed by placing the luc gene for firefly luciferase under the control of XylR and P u . When Escherichia coli cells were transformed with this plasmid vector, luminescence from the cells was induced in the presence of benzene, toluene, xylenes, and similar molecules. Accurate concentration dependencies of luminescence were obtained and exhibited K 1/2 values ranging from 39.0 ± 3.8 μM for 3-xylene to 2,690 ± 160 μM for 3-methylbenzylalcohol (means ± standard deviations). The luminescence response was specific for only toluene-like molecules that bind to and activate XylR. The biosensor cells were field tested on deep aquifer water, for which contaminant levels were known, and were able to accurately detect toluene derivative contamination in this water. The biosensor cells were also shown to detect BETX (benzene, toluene, and xylene) contamination in soil samples. These results demonstrate the capability of such a bacterial biosensor to accurately measure environmental contaminants and suggest a potential for its inexpensive application in field-ready assays.

Bibliography

Willardson, B. M., Wilkins, J. F., Rand, T. A., Schupp, J. M., Hill, K. K., Keim, P., & Jackson, P. J. (1998). Development and Testing of a Bacterial Biosensor for Toluene-Based Environmental Contaminants. Applied and Environmental Microbiology, 64(3), 1006â1012.

Authors 7

Barry M. Willardson (first)
Jon F. Wilkins (additional)
Timothy A. Rand (additional)
James M. Schupp (additional)
Karen K. Hill (additional)
Paul Keim (additional)
Paul J. Jackson (additional)

References 30 Referenced 141

10.1128/jb.171.12.6782-6790.1989
10.1016/S0021-9258(18)98483-8
10.1016/S0065-2911(08)60119-8
10.1128/aem.55.6.1323-1328.1989
10.1016/0378-1119(93)90344-3
10.1007/BF00265439
Harayama S. Timmis K. N. Aerobic biodegradation of aromatic hydrocarbons by bacteria Metal ions in biological systems Sigel H. Sigel A. 28 1992 99 156 Marcel Dekker Inc. New York, N.Y / Metal ions in biological systems / Aerobic biodegradation of aromatic hydrocarbons by bacteria by Harayama S. (1992)
10.1128/aem.60.5.1487-1494.1994
Hirayama K. Handbook of ultraviolet and visible absorption spectra of organic compounds. 1967 Plenum Press New York N.Y
Howard P. H. Handbook of environmental fate and exposure data for organic chemicals 1989 435 Lewis Publishers Chelsea Mich
10.1016/0925-4005(93)85353-C
10.1073/pnas.81.6.1688
10.1128/jb.163.3.863-869.1985
10.1073/pnas.84.15.5182
10.1016/0378-1119(88)90366-6
10.1016/S0022-2836(05)80317-1
10.1126/science.249.4970.778
10.1016/0968-0004(91)90163-P
10.1111/j.1365-2958.1993.tb01222.x
10.1016/S0021-9258(17)31696-4
10.1128/jb.166.1.9-14.1986
Schupp J. M. and P. Keim. Unpublished results.
Schupp J. M. Travis S. E. Price L. B. Shand R. F. Keim P. Rapid bacterial permeabilization reagent useful for enzyme assays. BioTechniques 19 1995 18 20 / BioTechniques / Rapid bacterial permeabilization reagent useful for enzyme assays. by Schupp J. M. (1995)
10.1128/jb.175.6.1596-1604.1993
10.1128/jb.176.6.1555-1560.1994
Spooner R. A. Lindsay K. Franklin F. C. H. Genetic, functional and sequence analysis of the xylR and xylS regulatory genes of the TOL plasmid pWWO. J. Gen. Microbiol. 132 1986 1347 1358 / J. Gen. Microbiol. / Genetic, functional and sequence analysis of the xylR and xylS regulatory genes of the TOL plasmid pWWO. by Spooner R. A. (1986)
Weast R. C. CRC handbook of chemistry and physics 60 1980 C–161 CRC Press Boca Raton, Fla / CRC handbook of chemistry and physics by Weast R. C. (1980)
Weast R. C. Grasselli J. G. Handbook of data on organic compounds 2nd ed. 1989 CRC Press Boca Raton Fla
10.1128/jb.124.1.7-13.1975
10.1073/pnas.79.3.874

Dates

Type	When
Created	5 years, 8 months ago (Dec. 19, 2019, 2:32 p.m.)
Deposited	3 years, 6 months ago (Feb. 23, 2022, 1:05 a.m.)
Indexed	1 month ago (July 30, 2025, 8:39 p.m.)
Issued	27 years, 6 months ago (March 1, 1998)
Published	27 years, 6 months ago (March 1, 1998)
Published Print	27 years, 6 months ago (March 1, 1998)

Funders 0

None

BibTeX

@article{Willardson_1998, title={Development and Testing of a Bacterial Biosensor for Toluene-Based Environmental Contaminants}, volume={64}, ISSN={1098-5336}, url={http://dx.doi.org/10.1128/aem.64.3.1006-1012.1998}, DOI={10.1128/aem.64.3.1006-1012.1998}, number={3}, journal={Applied and Environmental Microbiology}, publisher={American Society for Microbiology}, author={Willardson, Barry M. and Wilkins, Jon F. and Rand, Timothy A. and Schupp, James M. and Hill, Karen K. and Keim, Paul and Jackson, Paul J.}, year={1998}, month=mar, pages={1006–1012} }

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