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Appl Environ Microbiol, July 1998, p. 2730-2735, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Chromosomally Based tod-luxCDABE Whole-Cell Reporter for Benzene, Toluene, Ethybenzene, and Xylene (BTEX) Sensing

B. M. Applegate, S. R. Kehrmeyer, and G. S. Sayler*

Department of Microbiology and the Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee 37996-1605

Received 30 September 1997/Accepted 28 April 1998

A tod-luxCDABE fusion was constructed and introduced into the chromosome of Pseudomonas putida F1, yielding the strain TVA8. This strain was used to examine the induction of the tod operon when exposed to benzene, toluene, ethylbenzene, and xylene (BTEX) compounds and aqueous solutions of JP-4 jet fuel constituents. Since this system contained the complete lux cassette (luxCDABE), bacterial bioluminescence in response to putative chemical inducers of the tod operon was measured on-line in whole cells without added aldehyde substrate. There was an increasing response to toluene concentrations from 30 µg/liter to 50 mg/liter, which began to saturate at higher concentrations. The detection limit was 30 µg/liter. There was a significant light response to benzene, m- and p-xylenes, phenol, and water-soluble JP-4 jet fuel components, but there was no bioluminescence response upon exposure to o-xylene. The transposon insertion was stable and had no negative effect on cell growth.

* Corresponding author. Mailing address: Center for Environmental Biotechnology, 676 Dabney Hall, University of Tennessee---Knoxville, Knoxville, TN 37996-1605. Phone: (423) 974-8080. Fax: (423) 974-8086. E-mail: sayler{at}utk.edu.

Appl Environ Microbiol, July 1998, p. 2730-2735, Vol. 64, No. 7
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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