The methanogenic propionate degradation consortia were enriched in a propionate-fed semi-continuous bioreactor. The microbial community shift with depth, the microbial network and its correlation with metabolic pathway were also investigated. The results demonstrated that the maximum organic loading rate (OLR) of the reactor was 2.5 g propionic acid (HPr) L-1 d(-1) with approximately 1.20 LL-1 d(-1) of volumetric methane production (VMP). The organisms in the enrichment were spanning 36 bacterial phyla and 7 archaeal orders. Syntrophobacter, the main Hpr oxidizer in the digester, dominated bacteria with relative abundance changing from 63% to 37% with depth. The predominant methanogens shift from hydrogenotrophic Methanoculleus (similar to 60%) at the upper liquid layer to acetoclastic Methanothrix (similar to 51%) at the lower sediment layer in the bioreactor. These methanogens syntrophically support Syntrophobacter by degrading HPr catabolism by-products (H-2 and acetate). Other bacteria could scavenge anabolic products (carbohydrate and protein) presumably derived from detrital biomass produced by the HPr-degrading community.