An active microbiota, reaching up to 10 E+7 cells/g, was found to inhabit a naturally occurring asphalt lake characterized by low water activity and elevated temperature. Geochemical and molecular taxonomic approaches revealed novel and deeply branching microbial assemblages mediating anaerobic hydrocarbon degradation, metal respiration and C1 utilization pathways. These results open a window into the origin and adaptive evolution of microbial life within recalcitrant hydrocarbon matrices, and establish the site as a useful analog for the liquid hydrocarbon environments on Saturns moon Titan.