Applications of a colorimetric plate assay for soluble methane monooxygenase activity.
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.
Graham
D W
DW
Department of Civil Engineering and Engineering Mechanics, University of Arizona, Tucson 85721.
Korich
D G
DG
LeBlanc
R P
RP
Sinclair
N A
NA
Arnold
R G
RG
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
D004785:Environmental Pollutants; D010105:Oxygenases; C021555:methane monooxygenase