SOLUBLE METHANE MONOOXYGENASE PRODUCTION AND TRICHLOROETHYLENE DEGRADATION BY A TYPE-I METHANOTROPH, METHYLOMONAS-METHANICA-68-1

A methanotroph (strain 68-1), originally isolated from a trichloroethylene (TCE)-contaminated aquifer, was identified as the type I methanotroph Methylomonas methanica on the basis of intracytoplasmic membrane ultrastructure, phospholipid fatty acid profile, and 16S rRNA signature probe hybridization. Strain 68-1 was found to oxidize naphthalene and TCE via a soluble methane monooxygenase (sMMO) and thus becomes the first type I methanotroph known to be able to produce this enzyme. The specific whole-cell sMMO activity of 68-1, as measured by the naphthalene oxidation assay and by TCE biodegradation, was comparatively higher than sMMO activity levels in Methylosinus trichosporium OB3b grown in the same copper-free conditions. The maximal naphthalene oxidation rates of Methylomonas methanica 68-1 and Methylosinus trichosporium OB3b were 551 +/- 27 and 321 +/- 16 nmol h-1 mg of protein-1, respectively. The maximal TCE degradation rates of Methylomonas methanica 68-1 and Methylosinus trichosporium OB3b were 2,325 +/- 260 and 995 +/- 160 nmol h-1 mg of protein-1, respectively. The substrate affinity of 68-1 sMMO to naphthalene (K(m) 70 +/- 4 muM) and TCE (K(m) 225 +/- 13 muM), however, was comparatively lower than that of the sMMO of OB3b, which had affinities of 40 +/- 3 and 126 +/- 8 muM, respectively. Genomic DNA slot and Southern blot analyses with an sMMO gene probe from Methylosinus trichosporium OB3b showed that the sMMO genes of 68-1 have little genetic homology to those of OB3b. This result may indicate the evolutionary diversification of the sMMOs.