title: Bacterial oxygenases: in vivo enzyme biosensors for organic pollutants.
abstract: An in vivo enzyme-based biosensor platform was developed that uses specific oxygenase enzymes to detect aromatic compounds in water. Bacteria capable of degrading highly reduced hydrocarbons initiate substrate oxidation using well-characterised oxygenase enzymes, which due to their specificity, stability and high activity can be applied in vivo as biosensor components. Oxygenase enzyme activity was determined in vivo using BD Oxygen Biosensor plates to measure oxygenase-mediated oxygen depletion in the presence of specific aromatic analytes. The BTEX family of compounds (benzene, toluene, ethylbenzene and the three isomers of xylene) were used as model compounds. Detection limits and sensitivity achieved using this approach (microM detection range) was similar to levels achieved with oxygen electrode-based and some recombinant DNA-based approaches. No significant signals were detected with hydrocarbons that were not substrates of the initial oxygenases.
authors.affiliation: Lincoln Ventures Ltd., P.O. Box 133, Lincoln 8152, New Zealand.
authors.forename: Aynsley C
authors.initials: AC
authors.lastname: Tizzard
authors.forename: Gareth
authors.initials: G
authors.lastname: Lloyd-Jones
mesh_terms: D001420:Bacteria, Aerobic / Q000201:enzymology*; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004785:Environmental Pollutants / Q000032:analysis*; D006841:Hydrocarbons, Aromatic / Q000032:analysis* / Q000737:chemistry; D010100:Oxygen / Q000032:analysis*; D010105:Oxygenases / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004785:Environmental Pollutants; D006841:Hydrocarbons, Aromatic; D010105:Oxygenases; D010100:Oxygen