Molecular analysis of soluble methane monooxygenase and 16S rDNA from a type Ⅱmethanotroph

Abstract:

Abstract: The soluble methane monooxygenase (sMMO) from Methylosinus trichosporium IMV 3011 catalyzes the conversion of methane to methanol. [Objective] To identify the novel species Methylosinus trichosporium IMV 3011 and to describe its evolution status. [Methods]With the aid of the information from GenBank, we designed several sets of primers for PCR amplification and sequencing, the 16S rDNA and complete of genes sequence for soluble methane monooxygenases were gene sequenced and analyzed with biology software. [Results] We obtained a 5319 bp of full-length DNA of soluble methane monooxygenases and a 1290 bp of 16S rDNA. Software analysis for six open reading frames and the deduced amino acid sequences of soluble methane monooxygenases has shown that 99.0% to 82.7% identity to the counterpart of Methylosinus trichosporium OB3b, 99.4% to 81.8% identity and 99.8% to 89.2% similarity to the predicted amino acid of mmoX genes in compared five strains. The multiple alignments of MMOX amino acid residues reveal that there is high conservation in MMOX, especially in two Fe binding regions. [Conclusion] These results indicated that strain IMV 3011 should be a true member of Methylosinus trichosporium, and it is closer to the species Methylosinus trichosporium OB3b.