NITE Biological Resource Center (NBRC), National Institute of Technology and Evaluation (NITE), 2-5-8 Kazusakamatari, Kisarazu, Chiba 292-0818, Japan.
Department of Biological Sciences, Faculty of Science and Engineering, Chuo University, 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, Japan.
Int J Syst Evol Microbiol. 2011 Jan;61(Pt 1):138-143. doi: 10.1099/ijs.0.022723-0. Epub 2010 Feb 19.
Two methane-producing archaea, designated Mic5c12(T) and Mic6c05(T), were isolated from sludge deposited in a crude oil storage tank and a tubercle on the interior of a pipe transporting natural gas-containing brine, respectively. The isolates were Gram-staining-variable, non-motile rods and grew only on H(2)/CO(2). Strain Mic6c05(T) produced methane from some alcohols without showing any growth; strain Mic5c12(T) did not utilize alcohols. The optimum growth conditions for strain Mic5c12(T) were 35 °C, pH 6.5 and 0-0.68 M NaCl and for strain Mic6c05(T) were 40 °C, pH 6.0-7.5 and 0.34 M NaCl. Strain Mic5c12(T) was halotolerant and strain Mic6c05(T) was halophilic. Comparative 16S rRNA gene sequence analysis revealed that strains Mic5c12(T) and Mic6c05(T) belonged to the genus Methanobacterium and their closest relative was Methanobacterium subterraneum A8p(T) (97.3 and 97.9 % 16S rRNA gene sequence similarity, respectively). The findings from the 16S rRNA gene sequence analyses were supported by analysis of McrA, the alpha subunit of methyl-coenzyme M reductase. On the basis of phylogenetic analyses and phenotypic characteristics, two novel species are proposed, Methanobacterium petrolearium sp. nov. and Methanobacterium ferruginis sp. nov., with type strains Mic5c12(T) (=NBRC 105198(T) =DSM 22353(T)) and Mic6c05(T) (=NBRC 105197(T) =DSM 21974(T)), respectively.
两株产甲烷古菌,分别命名为 Mic5c12(T)和 Mic6c05(T),分别从储油罐中沉积的污泥和输送含盐水的管道内的结节中分离得到。分离株为革兰氏染色可变、非运动的杆状菌,仅在 H(2)/CO(2)上生长。菌株 Mic6c05(T)可以从一些醇类物质中产生甲烷,但没有表现出任何生长;菌株 Mic5c12(T)不能利用醇类物质。菌株 Mic5c12(T)的最佳生长条件为 35°C、pH 6.5 和 0-0.68 M NaCl,菌株 Mic6c05(T)的最佳生长条件为 40°C、pH 6.0-7.5 和 0.34 M NaCl。菌株 Mic5c12(T)为耐盐菌,菌株 Mic6c05(T)为嗜盐菌。比较 16S rRNA 基因序列分析表明,菌株 Mic5c12(T)和 Mic6c05(T)属于甲烷杆菌属,与 Methanobacterium subterraneum A8p(T)的亲缘关系最近(16S rRNA 基因序列相似度分别为 97.3%和 97.9%)。16S rRNA 基因序列分析的结果得到了 McrA(甲基辅酶 M 还原酶的 α 亚基)分析的支持。基于系统发育分析和表型特征,提出了两个新种,即石油甲烷杆菌(Methanobacterium petrolearium sp. nov.)和 Ferruginimethanobacterium ferruginis(Methanobacterium ferruginis sp. nov.),其模式菌株分别为 Mic5c12(T)=NBRC 105198(T)=DSM 22353(T)和 Mic6c05(T)=NBRC 105197(T)=DSM 21974(T)。
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