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A thermophilic strain ofMethanosarcina, designatedMethanosarcina strain TM-1, was isolated from a laboratory-scale 55°C anaerobic sludge digestor by the Hungate roll-tube technique. Penicillin andd -cycloserine, inhibitors of peptidoglycan synthesis, were used as selective agents to eliminate contaminating non-methanogens.Methanosarcina strain TM-1 had a temperature optimum for methanogenesis near 50°C and grew at 55°C but not at 60°C. Substrates used for methanogenesis and growth byMethanosarcina strain TM-1 were acetate (12-h doubling time), methanol (7- to 10-h doubling time), methanol-acetate mixtures (5-h doubling time), methylamine, and trimethylamine. When radioactively labeled acetate was the sole methanogenic substrate added to the growth medium, it was predominantly split to methane and carbon dioxide. When methanol was also present in the medium, the metabolism of acetate shifted to its oxidation and incorporation into cell material. Electrons derived from acetate oxidation apparently were used to reduce methanol. H2 -CO2 was not used for growth and methanogenesis byMethanosarcina strain TM-1. When presented with both H2 -CO2 and methanol,Methanosarcina strain TM-1 was capable of limited hydrogen metabolism during growth on methanol, but hydrogen metabolism ceased once the methanol was depleted.Methanosarcina strain TM-1 required a growth factor (or growth factors) present in the supernatant of anaerobic digestor sludge. Growth factor requirements and the inability to use H2 -CO2 are characteristics not found in other describedMethanosarcina strains. The high numbers ofMethanosarcina -like clumps in sludges from thermophilic digestors and the fast generation times reported here forMethanosarcina TM-1 indicate thatMethanosarcina may play an important role in thermophilic methanogenesis.

Isolation and Characterization of a Thermophilic Strain of Methanosarcina Unable to Use H 2 -CO 2 for Methanogenesis

Isolation and Characterization of a Thermophilic Strain of Methanosarcina Unable to Use H ₂ -CO ₂ for Methanogenesis