Abstract
Two strains of Methanosarcina (M. Barkeri strain MS, isolated from sewage sludge, and strain UBS, isolated from lake sediments) were found to have similar cellular properties and to have DNA base compositions of 44 mol percent guanosine plus cytosine. Strain MS was selected for further studies of its one-carbon metabolism. M. barkeri grew autotrophically via H2 oxidation/CO2 reduction. The optimum temperature for growth and methanogenesis was 37°C. H2 oxidation proceeded via an F420-dependent NADP+-linked hydrogenase. A maximum specific activity of hydrogenase in cell-free extracts, using methyl viologen as electron acceptor, was 6.0 μmol min · mg protein at 37°C and the optimum pH (9.0). M. barkeri also fermented methanol andmethylamine as sole energy sources for growth. Cell yields during growth on H2/CO2 and on methanol were 6.4 and 7.2 mg cell dry weight per mmol CH4 formed, respectively. During mixotrophic growth on H2/CO2 plus methanol, most methane was derived from methanol rather than from CO2. Similar activities of hydrogenase were observed in cell-free extracts from H2/CO2-grown and methanol-grown cells. Methanol oxidation apparently proceeded via carrierbound intermediates, as no methylotrophy-type of methanol dehydrogenase activity was observed in cell-free extracts. During growth on methanol/CO2, up to 48% of the cell carbon was derived from methanol indicating that equivalent amounts of cell carbon were derived from CO2 and from an organic intermediate more reduced than CO2. Cell-free extracts lacked activity for key cell carbon synthesis enzymes of the Calvin cycle, serine path, or hexulose path.
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Abbreviations
- CAPS:
-
cycloaminopropane sulfonic acid
- CH3-SCoM:
-
methyl coenzyme M
- DCPIP:
-
2,6-dichlorophenolindophenol
- DEAE:
-
diethylaminoethyl
- dimethyl POPOP:
-
1,4-bis-2-(4-mothyl-5-phenyloxazolyl)-benzene
- DNA:
-
deoxyribonucleic acid
- dpm:
-
dismtegrations per min
- DTT:
-
dithiothreitol
- EDTA:
-
ethylenediamine tetraacetic acid
- F420 :
-
factor 420
- G+C:
-
guanosine plus cytosine
- NAD+ :
-
nicotinamide adenine dinucleotide
- NADP+ :
-
nicotinamide adenine dinucleotide phosphate
- PBBW:
-
phosphate buffered basal Weimer
- PMS:
-
phenazine methosulfate
- PPO:
-
2,5-diphenyloxazole
- rRNA:
-
ribosomal ribonucleic acid
- RuBP:
-
ribulose-1,5-bisphosphate
- Tris:
-
tris-hydroxymethyl-aminomethane
- μmax:
-
maximum specific growth rate
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Weimer, P.J., Zeikus, J.G. One carbon metabolism in methanogenic bacteria. Arch. Microbiol. 119, 49–57 (1978). https://doi.org/10.1007/BF00407927
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DOI: https://doi.org/10.1007/BF00407927