Anaerobic digestion is one of the most economical ways to treat organic wastes; the production of excessive sludge is limited; aeration is not required; operation cost is low; valuable energy resource (methane) is produced. However, propionate, an intermediate substance during the process, is known to inhibit biochemical process at high concentration; but it is difficult to be degraded biologically as its oxidation is thermodynamically unfavorable (△G>0). However, there are several microorganisms which could oxidize propionate fully by their syntrophic association. Therefore, in this research, the propionate enrichment culture was produced in large amount (~1L), and the main players which are responsible for utilizing propionate are identified. Furthermore, the kinetic parameter was obtained from this research. Seed came from two different full-scale anaerobic digesters which is located in Joongrang (JR) and Sincheon (SC). 16s rRNA gnee based metagenomics results indicated that Smithella propionica and Methanobacterium beijingense were the most dominant species in all seeds. The fact that bacterial family Syntrophomonadaceae and archaeal genus Methanosaeta were also found significantly in all seeds means that there might be symbiotic relationship among propionate oxidizing bacteria, butyrate oxidizing bacteria, hydrogenotrophic methanogens and acetotrophic methanogens in complete oxidation of propionate. Based on the first order kinetic which indicated propionate degrading rate, k (1/day) on JR and SC was represented 0.0243 and 0.0223, respectively. Also, based on Modified-Gompertz model, umax on JR and SC was represented as 0.024 and 0.016, respectively.