Activity-dependent labeling of oxygenase enzymes in a trichloroethene-contaminated groundwater site.
A variety of naturally occurring bacteria produce enzymes that cometabolically degrade trichloroethene (TCE), including organisms with aerobic oxygenases. Groundwater contaminated with TCE was collected from the aerobic region of the Test Area North site of the Idaho National Laboratory. Samples were evaluated with enzyme activity probes, and resulted in measurable detection of toluene oxygenase activity (6-79% of the total microbial cells). Wells from both inside and outside contaminated plume showed activity. Toluene oxygenase-specific PCR primers determined that toluene-degrading genes were present in all groundwater samples evaluated. In addition, bacterial isolates were obtained and possessed toluene oxygenase enzymes, demonstrated activity, and were dominated by the phylotype Pseudomonas. This study demonstrated, through the use of enzymatic probes and oxygenase gene identification, that indigenous microorganisms at a contaminated site were cometabolically active. Documentation such as this can be used to substantiate observations of natural attenuation of TCE-contaminated groundwater plumes.
Lee
M Hope
MH
Idaho State University, Department of Biological Sciences, Pocatello, ID 83209, USA. hlee@northwind-inc.com
Clingenpeel
Scott C
SC
Leiser
Owen P
OP
Wymore
Ryan A
RA
Sorenson
Kent S
KS
Watwood
Mary E
ME
D001419:Bacteria / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D001673:Biodegradation, Environmental; D017931:DNA Primers; D004463:Ecology / Q000379:methods; D005618:Fresh Water; D005798:Genes, Bacterial; D007061:Idaho; D010105:Oxygenases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D014241:Trichloroethylene / Q000378:metabolism*; D014874:Water Pollutants, Chemical / Q000378:metabolism*
D017931:DNA Primers; D014874:Water Pollutants, Chemical; D014241:Trichloroethylene; D010105:Oxygenases