The microbial origin of methane produced from sedimentary basins is a subject of great interest, with implications for the global cycling of carbon as well as natural gas exploration. Despite the growing body of research in sedimentary basin methanogenesis, few studies have sought to understand the subsurface microbial communities that produce methane, the metabolic pathways involved in the decomposition of ancient organic matter, or the components of ancient organic matter that are consumed. This research examined shale-derived dissolved organic matter (DOM) as a potential substrate to support a subsurface methanogenic community in a known microbial shale gas reserve, the Antrim Shale in the Michigan Basin, USA. Experiments were conducted that enriched fermentative and sulfate-reducing microbial communities from Antrim Shale formation waters. Additionally, 1H NMR spectroscopy was used to characterize shale-derived DOM solutions before and after they were used as growth media for fermentative and sulfate-reducing microbial communities, and to characterize the DOM of the Antrim Shale formation waters. The results of the enrichment studies demonstrate that both fermentative and sulfate-reducing microbial communities from the Antrim Shale are capable of growth using shale-derived DOM as their only source of organic carbon; further, the production of methane in a fermentative enrichment demonstrates that methanogenesis can be supported by shale-derived DOM alone. The 1H NMR characterization studies of the shale-derived DOM solutions before and after growth revealed subtle but detectable differences in DOM compositions, indicating the production and consumption of DOM components by the fermentative and sulfate-reducing microbial communities. Characterization analyses of Antrim Shale formation waters suggest that salinity and microbiological activity may influence the liberation of aliphatic and aromatic compounds from shale. The DOM characterization studies also suggest that carboxylic acids may be consumed by methanogenic communities in the Antrim Shale, and aromatic compounds may be produced by the enriched microbial communities and the communities present in the Antrim Shale.