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KMID : 0545120020120050819
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Journal of Microbiology and Biotechnology
2002 Volume.12 No. 5 p.819 ~ p.825
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Isolation and Molecular Analysis of Methanol Oxidation Genes in an Obligate Methylotrophic Bacterium, Methylobacillus sp. Strain SK-5
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CHOI, HACK-SUN
KIM, JIN-KWON/AHN, YEONGHEE/KOH, MOONJOO/KIM, SI WOUK
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Abstract
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Methanol dehydrogenase (MDH) is a key enzyme in the process of methanol oxidation in methylotrophic bacteria. However, information in MDH genes from genus Methylobacillus is limited. In this study, a 6.5-kb HindⅢ DNA fragment of Methylobacillus sp. SK-5 chromosomal DNA was isolated from the genomic library of the strain by using a degenerate oligonucleotide probe that was designed based on N-terminal amino acid sequence of the MDH α subunit purified from the strain. Molecular analysis of the fragment revealed four tightly clustered genes (mxaFJGI) involved in the methanol oxidation. The first and fourth genes were very similar to mxaF(77% identity for nucleotides and 78% identity for amino acids) ands mxaI(67% identity for nucleotides and 68% identity for amino acids) genes, respectively, from Methylovorus sp. SS1. Genes mxaF and mxaI encode α and β subunits of MDH, respectively. The two subunits were identified from purified MDH from Methylobacillus sp. SK-5. A dendrogram constructed by comparison of amino acid sequences of MDH α subunits suggests that MxaF from Methylobacillus sp. SK-5 belongs to a subfamily cluster of MDH α subunits from β-subgroup Proteobacteria. The subfamily cluster is separated from the other subfamily that consists of α- and γ-subgroup Proteobacteria. This study provided information on MDH genes from a methylotrophic bacterium in β-subgroup Proteobacteria, which would aid to better develop a gene probe to detect one-carbon metabolizing bacteria.
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