This study examined the significance of propionate to acetate (HPr/HAc) ratios on microbial syntrophy and competition in microbial electrolysis cell-assisted anaerobic digestion (MEC-AD). In addition to molecular biology and phylogenetic analysis, a numerical MEC-AD model was developed by modifying Anaerobic Digestion Model No.1 to predict the effects of different HPr/HAc ratios (0.5, 1.5, 2.5, and 5). The HPr/HAc ratios of 0.5 and 1.5 maintained efficient syntrophy among electroactive bacteria, hydrogenotrophic methanogens, and homoacetogens, leading to higher methane yields. In contrast, higher HPr/HAc ratios of 2.5 and 5 were detrimental to methanogenesis. Both microbial community analysis and numerical modeling results suggested that higher propionate levels could promote the enrichment of H2-utilizing acetogens, thereby triggering their competition with hydrogenotrophic methanogens. Moreover, protein fraction in extracellular polymeric substances and the relative expression of genes associated with extracellular electron transfer in both anode and cathode biofilms were markedly decreased with increasing HPr/HAc ratios, indicating partial inhibition of microbial electroactivity. Overall, these results illuminate deep insight into anaerobic syntrophy, contributing to the process kinetics and methane yields in MEC-AD systems. Furthermore, from a practical viewpoint, the results can also be helpful in effective control of MEC-AD operation without propionate accumulation.