Methanogenic fermentation of benzoate in an enrichment culture

Enrichment cultures inoculated with black mud fermented benzoate according to the stoichiometric equation: 4 C6H5CO2H+18 H2O → 15 CH4+13 CO2.Trans-2-hydroxycyclohexanecarboxylate, 2-oxo-cyclohexanecarboxylate, pimelate, caproate, butyrate, acetate, and molecular hydrogen were shown to be regular components of the culture fluid occurring in low concentrations. Inhibition of methanogenesis by chloroform, 4-chlorobutyrate, or 2-bromooctanoate resulted in a cessation of the benzoate breakdown after all intermediates had accumulated. It is proposed that benzoate is fermented via a direct reductive pathway to butyrate, acetate, H2, and CO2, whereafter butyrate is converted to acetate and H2, and the latter substrates are fermented to CH4 and CO2 by methane producers.