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Isolation and Characterization of Methanolobus bombayensis sp. nov., a Methylotrophic Methanogen That Requires High Concentrations of Divalent Cations

¹Department of Chemistry, Biochemistry, and Molecular Biology, Oregon Graduate Institute of Science & Technology, Portland, Oregon 97291-1000
²Department of Microbiology, Agharkar Research Institute, Pune 411004, India
³Department of Microbiology, University of Illinois, Urbana, Illinois 61801
⁴ Department of Environmental Science and Engineering, Oregon Graduate Institute of Science & Technology, Portland, Oregon 97291-1000

^* Corresponding author. Mailing address: Department of Environmental Science and Engineering, Oregon Graduate Institute, P.O. Box 91000, Portland, OR 97291-1000. Phone: (503) 690-1146. Fax: (503) 690-1273. Electronic mail address: boone{at}ese.ogi.edu.

ABSTRACT

Methanolobus bombayensis B-1^T (= OCM 438^T) (T = type strain) was isolated from Arabian Sea sediments obtained near Bombay, India. This strain grew on methylamines, methanol, and dimethyl sulfide, but it did not catabolize H₂-CO₂, acetate, or formate. The cells were nonmotile, irregular coccoids (diameter, 1.0 to 1.5 µm) and occurred singly. Electron micrographs revealed that a cell membrane and a protein cell wall were present. The cells grew fastest at mesophilic temperatures, at a neutral pH, and at salinity levels near the salinity level of the ocean, and they required about 30 mM divalent cations (Mg²⁺ and Ca²⁺). The cells grew in mineral medium, but growth was greatly stimulated by yeast extract and peptones. The guanine-plus-cytosine content of the DNA was 39.2 ± 0.1 mol%. A comparison of 16S rRNA sequences showed that strain B-1^T was phylogenetically related to Methanolobus vulcani, but the sequences of these organisms differed by 2%.

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