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Methanosarcina sp. strain TM-1 and Methanosarcina acetivorans produced and consumed H2 to maintain H2 partial pressures of 16 to 92 Pa in closed cultures during growth on acetate. Strain TM-1 produced H2 continuously when H2 was continuously removed from the culture. The potential physiological significance of H2 in acetate metabolism to methane is discussed.