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Applications of a colorimetric plate assay for soluble methane monooxygenase activity.

机译:比色板测定法对可溶性甲烷单加氧酶活性的应用。

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摘要

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.
机译:描述了一种直接方法,用于筛选甲烷营养型菌落在固体培养基上的可溶性甲烷单加氧酶(sMMO)活性。这种活性导致1-萘酚(当sMMO与萘反应时形成)与邻二苯胺(四唑化)之间形成有色络合物。表达sMMO的甲烷营养菌落在依次暴露于萘和邻二苯胺啶后变成深紫色。该方法是在两个潜在应用程序的上下文中进行评估的。第一个方法是在专用于氯乙烯处理的甲烷修正的不饱和土壤柱中枚举三甲基甲烷菌OB3b。第二个应用是从卫生垃圾填埋场土壤中分离和计数含sMMO的甲烷营养生物。该技术在两种应用中均有效。

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