Applications of a colorimetric plate assay for soluble methane monooxygenase activity

D. W. Graham, D. G. Korich, R. P. LeBlanc, N. A. Sinclair, Robert G Arnold

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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.

Original languageEnglish (US)
Pages (from-to)2231-2236
Number of pages6
JournalApplied and Environmental Microbiology
Volume58
Issue number7
StatePublished - 1992

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ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

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