Optimization of trichloroethylene oxidation by methanotrophs and the use of a colorimetric assay to detect soluble methane monooxygenase activity

Gregory A. Brusseau, Hsien Chyang Tsien, Richard S. Hanson, Lawrence P Wackett

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173 Scopus citations

Abstract

Methylosinus trichosporium OB3b biosynthesizes a broad specificity soluble methane monooxygenase that rapidly oxidizes trichloroethylene (TCE). The selective expression of the soluble methane monooxygenase was followed in vivo by a rapid colorimetric assay. Naphthalene was oxidized by purified soluble methane monooxygenase or by cells grown in copper-deficient media to a mixture of 1-naphthol and 2-naphthol. The naphthols were detected by reaction with tetrazotized o-dianisidine to form purple diazo dyes with large molar absorptivities. The rate of color formation with the rapid assay correlated with the velocity of TCE oxidation that was determined by gas chromatography. Both assays were used to optimize conditions for TCE oxidation by M. trichosporium OB3b and to test several methanotrophic bacteria for the ability to oxidize TCE and naphthalene.

Original languageEnglish (US)
Pages (from-to)19-29
Number of pages11
JournalBiodegradation
Volume1
Issue number1
DOIs
StatePublished - Mar 1 1990

Keywords

  • Methylosinus trichosporium
  • methane monooxygenase
  • methanotroph
  • naphthalene
  • trichloroethylene

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