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Detection of dichloromethane with a bioluminescent (lux) bacterial bioreporter

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The focus of this research effort was to develop an autonomous, inducible, lux-based bioluminescent bioreporter for the real-time detection of dichloromethane. Dichloromethane (DCM), also known as methylene chloride, is a volatile organic compound and one of the most commonly used halogenated solvents in the U.S., with applications ranging from grease and paint stripping to aerosol propellants and pharmaceutical tablet coatings. Predictably, it is released into the environment where it contaminates air and water resources. Due to its classification as a probable human carcinogen, hepatic toxin, and central nervous system effector, DCM must be carefully monitored and controlled. Methods for DCM detection usually rely on analytical techniques such as solid-phase microextraction (SPME) and capillary gas chromatography or photoacoustic environmental monitors, all of which require trained personnel and/or expensive equipment. To complement conventional monitoring practices, we have created a bioreporter for the self-directed detection of DCM by taking advantage of the evolutionary adaptation of bacteria to recognize and metabolize chemical agents. This bioreporter, Methylobacterium extorquens DCM lux , was engineered to contain a bioluminescent luxCDABE gene cassette derived from Photorhabdus luminescens fused downstream to the dcm dehalogenase operon, which causes the organism to generate visible light when exposed to DCM. We have demonstrated detection limits down to 1.0 ppm under vapor phase exposures and 0.1 ppm under liquid phase exposures with response times of 2.3 and 1.3 h, respectively, and with specificity towards DCM under relevant industrial environmental monitoring conditions.

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Acknowledgments

Research support was provided by the NASA Advanced Environmental Monitoring and Control Program under cooperative agreement number NNJ04HF02A and the National Science Foundation Division of Biological Infrastructure (DBI) under award number DBI-0963854.

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Authors and Affiliations

  1. The Center for Environmental Biotechnology, University of Tennessee, 676 Dabney Hall, Knoxville, TN, 37996, USA

    Nicholas Lopes, Shawn A. Hawkins, Patricia Jegier, Fu-Min Menn, Gary S. Sayler & Steven Ripp

  2. The Department of Biosystems Engineering and Soil Sciences, University of Tennessee, 676 Dabney Hall, Knoxville, TN, 37996, USA

    Shawn A. Hawkins

  3. The Joint Institute for Biological Sciences, Oak Ridge National Laboratory, Oak Ridge, TN, 37831, USA

    Fu-Min Menn & Gary S. Sayler

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  1. Nicholas Lopes
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  2. Shawn A. Hawkins
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  3. Patricia Jegier
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  4. Fu-Min Menn
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  5. Gary S. Sayler
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  6. Steven Ripp
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Correspondence to Steven Ripp.

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Lopes, N., Hawkins, S.A., Jegier, P. et al. Detection of dichloromethane with a bioluminescent (lux) bacterial bioreporter. J Ind Microbiol Biotechnol 39, 45–53 (2012). https://doi.org/10.1007/s10295-011-0997-5

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  • DOI: https://doi.org/10.1007/s10295-011-0997-5

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