title: Detection of dichloromethane with a bioluminescent (lux) bacterial bioreporter.
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.
authors.affiliation: The Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN 37996, USA.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Lopes
authors.forename: Shawn A
authors.initials: SA
authors.lastname: Hawkins
authors.forename: Patricia
authors.initials: P
authors.lastname: Jegier
authors.forename: Fu-Min
authors.initials: FM
authors.lastname: Menn
authors.forename: Gary S
authors.initials: GS
authors.lastname: Sayler
authors.forename: Steven
authors.initials: S
authors.lastname: Ripp
mesh_terms: D004784:Environmental Monitoring / Q000379:methods; D017930:Genes, Reporter; D008163:Luminescent Measurements; D008752:Methylene Chloride / Q000032:analysis*; D020601:Methylobacterium extorquens / Q000235:genetics* / Q000378:metabolism; D009876:Operon; D020637:Photorhabdus / Q000235:genetics*; D012997:Solvents / Q000032:analysis*
chemical_list: D012997:Solvents; D008752:Methylene Chloride