Anaerobic fermentation of fructose in a mixed culture of fermentative and methanogenic bacteria

Summary Two newly isolated strains of Methanosarcina, strains JKAD and DALS, were grown in monoculture and in mixed culture in combination with Acetobacterium woodii WB1. Methanosarcina strains convert acetate into methane and carbon dioxide while Acetobacterium woodii grows on fructose, producing acetate via homoacetate fermentation. Monocultures of A. woodii in continuous culture consumed up to 6 mmoles g^-1 dry weight (dw) h^-1 of fructose and produced up to 12.9 mmoles g^-1 dw h^-1 of acetate at a dilution rate (D) of 0.13 h^-1. In batch growth the methanogenic bacteria produced up to 12.1 mmoles g^-1 dw h^-1 of CH₄ at a specific growth rate of 0.043 h^-1. In continuous cultivation the specific growth rate and the specific methane production of Methanosarcina were lower than in batch cultures, with values of 0.031 h^-1 and 3.1 mmoles g^-1 dw h^-1 of methane, respectively. In combination, A. woodii and Methanosarcina strain DALS in batch cultures completely converted fructose to methane and carbon dioxide with a maximum specific methane production rate of 1.9 mmoles g^-1 dw h^-1 of methane. In continuous cultivation these mixed cultures produced between 1.2 and 2 mmoles g^-1 dw h^-1 of CH₄ at a dilution rate of up to 0.043 h^-1. The methanogens were washed out at D values higher than 0.043 h^-1 for A. woodii and Methanosarcina strain JKAD, and higher than 0.05 h^-1 for A. woodii and Methanosarcina strain DALS. Data obtained from defined mixed cultures allow one to follow interactions in a mixed population of two species with different growth constants.