title: Applications of a colorimetric plate assay for soluble methane monooxygenase activity.
abstract: A straightforward method is described for screening methanotrophic colonies for soluble methane monooxygenase (sMMO) activity on solid media. Such activity results in the development of a colored complex between 1-naphthol, which is formed when sMMO reacts with naphthalene, and o-dianisidine (tetrazotized). Methanotrophic colonies expressing sMMO turned deep purple when exposed successively to naphthalene and o-dianisidine. The method was evaluated within the contexts of two potential applications. The first was for the enumeration of Methylosinus trichosporium OB3b in a methane-amended, unsaturated soil column dedicated to vinyl chloride treatment. The second application was for the isolation and enumeration of sMMO-bearing methanotrophs from sanitary landfill soils. The technique was effective in both applications.
authors.affiliation: Department of Civil Engineering and Engineering Mechanics, University of Arizona, Tucson 85721.
authors.forename: D W
authors.initials: DW
authors.lastname: Graham
authors.forename: D G
authors.initials: DG
authors.lastname: Korich
authors.forename: R P
authors.initials: RP
authors.lastname: LeBlanc
authors.forename: N A
authors.initials: NA
authors.lastname: Sinclair
authors.forename: R G
authors.initials: RG
authors.lastname: Arnold
mesh_terms: D001673:Biodegradation, Environmental; D003124:Colorimetry / Q000379:methods*; D004785:Environmental Pollutants / Q000378:metabolism; D008772:Methylococcaceae / Q000201:enzymology / Q000302:isolation & purification; D010105:Oxygenases / Q000032:analysis*; D012988:Soil Microbiology; D012995:Solubility
chemical_list: D004785:Environmental Pollutants; D010105:Oxygenases; C021555:methane monooxygenase