Methanogenesis from sucrose by defined immobilized consortia.

A bacterial consortium capable of sucrose degradation primarily to CH(4) and CO(2) was constructed, with acetate as the key methanogenic precursor. In addition, the effect of agar immobilization on the activity of the consortium was determined. The primary fermentative organism, Escherichia coli, produced acetate, formate, H(2), and CO(2) (known substrates for methanogens), as well as ethanol and lactate, compounds that are not substrates for methanogens. Oxidation of the nonmethanogenic substrates, lactate and ethanol, to acetate was mediated by the addition of Acetobacterium woodii and Desulfovibrio vulgaris. The methanogenic stage was accomplished by the addition of the acetophilic methanogen Methanosarcina barkeri and the hydrogenophilic methanogen Methanobacterium formicicum. Results of studies with low substrate concentrations (0.05 to 0.2% [wt/vol]), a growth-limiting medium, and the five-component consortium indicated efficient conversion (40%) of sucrose carbon to CH(4). Significant decreases in yields of CH(4) and rates of CH(4) production were observed if any component of the consortium was omitted. Approximately 70% of the CH(4) generated occurred via acetate. Agar-immobilized cells of the consortium exhibited yields of CH(4) and rates of CH(4) production from sucrose similar to those of nonimmobilized cells. The rate of CH(4) production decreased by 25% when cysteine was omitted from reaction conditions and by 40% when the immobilized consortium was stored for 1 week at 4 degrees C.