Development and testing of a bacterial biosensor for toluene-based environmental contaminants

Barry M. Willardson, Jon F. Wilkins, Timothy A. Rand, James M. Schupp, Karen K. Hill, Paul S Keim, Paul J. Jackson

Research output: Contribution to journalArticle

125 Citations (Scopus)

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 XyIR 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( 1/4 ) 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 XyIR. 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.

Original languageEnglish (US)
Pages (from-to)1006-1012
Number of pages7
JournalApplied and Environmental Microbiology
Volume64
Issue number3
StatePublished - Mar 1998

Fingerprint

biosensors
toluene
Toluene
Biosensing Techniques
pollution
plasmid
xylene
pollutant
luminescence
Plasmids
benzene
Luminescence
Benzene
Xylenes
plasmids
testing
water pollution
Water
Soil
cells

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Willardson, B. M., Wilkins, J. F., Rand, T. A., Schupp, J. M., Hill, K. K., Keim, P. S., & Jackson, P. J. (1998). Development and testing of a bacterial biosensor for toluene-based environmental contaminants. Applied and Environmental Microbiology, 64(3), 1006-1012.

Development and testing of a bacterial biosensor for toluene-based environmental contaminants. / Willardson, Barry M.; Wilkins, Jon F.; Rand, Timothy A.; Schupp, James M.; Hill, Karen K.; Keim, Paul S; Jackson, Paul J.

In: Applied and Environmental Microbiology, Vol. 64, No. 3, 03.1998, p. 1006-1012.

Research output: Contribution to journalArticle

Willardson, BM, Wilkins, JF, Rand, TA, Schupp, JM, Hill, KK, Keim, PS & Jackson, PJ 1998, 'Development and testing of a bacterial biosensor for toluene-based environmental contaminants', Applied and Environmental Microbiology, vol. 64, no. 3, pp. 1006-1012.
Willardson, Barry M. ; Wilkins, Jon F. ; Rand, Timothy A. ; Schupp, James M. ; Hill, Karen K. ; Keim, Paul S ; Jackson, Paul J. / Development and testing of a bacterial biosensor for toluene-based environmental contaminants. In: Applied and Environmental Microbiology. 1998 ; Vol. 64, No. 3. pp. 1006-1012.
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