The reaction pathway for the formation of methane from acetate was investigated in sludge from 13 different biogas reactors. By following the conversion of [2‐14C]acetate and [14C]bicarbonate it was shown that methane formation by syntrophic acetate oxidation was the dominating mechanism for acetotrophic methanogenesis in sludge containing high levels of salts, mainly ammonium, and volatile fatty acids. In one biogas reactor the degree of syntrophic acetate oxidation increased as the concentrations of salts and volatile fatty acids increased. Statistical evaluation with principal component analysis showed a close correlation between the degree of syntrophic acetate oxidation and the concentrations of ammonium and potassium. The acetate degradation rate was lower (10–800 times) in biogas reactors in which methane was formed through syntrophic acetate oxidation than in biogas reactors where acetate was directly cleaved to methane and carbon dioxide. Microscopic observations revealed a predominance of one type of methanogen in the reactors with syntrophic acetate oxidation. Isolation and characterization, including substrate spectrum, mol% G+C, polyamine and 16S rRNA analysis, showed that the strains were hydrogenotrophic methanogens belonging to the genus Methanoculleus.

Two new methanogenic bacteria, Methanocorpusculum sinense spec. nov. strain DSM 4274 from a pilot plant for treatment of distillery wastewater in Chengdu (Province Sichuan, China), and Methanocorpusculum bavaricum spec. nov. strain DSM 4179, from a wastewater pond of the sugar factory in Regensburg (Bavaria, FRG) are described. Methanocorpusculum strains are weakly motile and form irregularly coccoid cells, about 1 micron in diameter. The cell envelope consists of a cytoplasmic membrane and a S-layer, composed of hexagonally arranged glycoprotein subunits with molecular weights of 90,000 (Methanocorpusculum parvum), 92,000 (M. sinense), and 94,000 (M. bavaricum). The center-to-center spacings are 14.3 nm, 15.8 nm and 16.0 nm, respectively. Optimal growth of strains is obtained in the mesophilic temperature range and at a pH around 7. Methane is produced from H2/CO2, formate, 2-propanol/CO2 and 2-butanol/CO2 by M. parvum and M. bavaricum, whereas M. sinense can only utilize H2/CO2 and formate. Growth of M. sinense and M. bavaricum is dependent on the presence of clarified rumen fluid. The G + C content of the DNA of the three strains is ranging from 47.7-53.6 mol% as determined by different methods. A similar, but distinct polar lipid pattern indicates a close relationship between the three Methanocorpusculum species. The polyamine patterns of M. parvum, M. sinense and M. bavaricum are similar, but distinct from those of other methanogens and are characterized by a high concentration of the otherwise rare 1,3-diaminopropane. Quantitative comparison of the antigenic fingerprint of members of Methanocorpusculum revealed no antigenic relationship with any one of the reference methanogens tested. On the basis of the distant phylogenetic position of M. parvum and the data presented in this paper a new family, the Methanocorpusculaceae fam. nov., is defined.

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