• Korean Journal of Microbiology
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• v.29 no.4
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• pp.250-257
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• 1991

An obligate methanol-oxidizing bacterium, Methylobacillus sp. strain SK1, which grows only on methanol was isolated from soil. The isolate was nonmotile Gram-negtive rod. It does not have internal membrane system. The colonies were small, whitish-yellow, and smooth. The guanine plus cytosine content of the DNA was 48 mol%. Cellular fatty acids consisted predominantly of large amounts of straight-chain saturated $C_{16:0}$ acid and unsaturated $C_{16:1}$ acid. The major ubiquinone was Q-8, and Q-10 was present as minor component. The cell was obligately aerobic and exhibited catalase, but no oxidase, activity. Poly-.betha.-hydroxybutyrate, endospores, or cysts were not observed. the isolate could grow only on methanol in mineral medium. Growth factors were not required. The isolate was unable to use methane, formaldehyde, formate, methylamine, and several other organic compounds tested as a sole source of carbon and energy. Growth was optimal at 35.deg.C and pH 7.5. It could not grow at 42.deg.C. The doubling time was 1.2h at 30.deg.C when grown with 1.0%(v/v) methanol. The growth was not affected by antibiotics inhibiting cell wall synthesis and carbon monoxide but was completely suppressed by those inhibiting protein synthesis. Methanol was found to be assimilated through the ribulose monophosphate pathway. Cytochromes of b-, c-, and o- types were found. Cell-free extracts contained a phenazine methosulfate-linked methanol dehydrogenase activity, which required ammonium ions as an activator. Cells harvested after the late exponential phase seemed to contain blue protein.ein.

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.261-266
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• 1993

A new methyltrophic bacterium which utilizes methanol as a sole source of carbon and energy was isolated from soil. It was Gram-negative, nonmotile, nonspore-forming rod, and strictly aerobic bacterium. Catalase and oxidase activities were present. Nitrate was reduced to nitrite. Vitamins and other growth factors were not required. Generation time was 1.6 hr under the optimal condition. The isolate assimilated methanol via the ribulose mono-phosphate pathway (Enter-Doudoroff varient) and did not have .alpha.-ketoglutarate dehydrogenase. It assimilated ammonia through glutamate dehydrogenase. The guanine plus cytosine content of the DNA was 61.0 mol%. The celular fatty acid composition was primarily straight-chain saturated $C^{16 : 0}$ acids (palmitic acids) and unsaturated $C_{16 :1}$ acid (palmitoleic acids), and the isolate also contained two unidentified $C_{17}$ branched fatty acids. The major ubiquinone was Q-8, and Q-6 and Q-7 were present as minor components. Phosphatidylethanolamine and phosphatidylglycerol were predominantly present, and diphosphatidyglycerol was also detected. Based on the physiological and biochemical properties, the isolate was assigned to a novel species of the genus Methylobacillus, Methylobacillus methanolovorus sp. nov.

• Molecules and Cells
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• v.23 no.3
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• pp.370-378
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• 2007

A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.