This study evaluated the efficacy of acclimated cow manure as a seed microbiome to enhance food waste hydrolysis. Anaerobic hydrolysis was performed on simulated food waste in a hydrolytic–acidogenic leach bed reactor (LBR) operated in batch mode under mesophilic conditions (35 °C) for 16 days. The acclimation process involved three sequential runs: Run-1 utilized 20% (w/w) cow manure as seed, Run-2 employed the digestate from Run-1 (day 5), and Run-3 used the digestate from Run-1 (day 10). Run-3 achieved 70.4% removal of volatile solids (VSs), surpassing Run-1 (47.1%) and Run-2 (57.1%). Compared with the first run, the production of chemical oxygen demand (COD) and total soluble products (TSPs) increased by 48.7% and 75.9%, respectively, in Run-3. The hydrolysis rate of proteins was 48.4% in Run-1, while an increase of 16.9% was achieved in Run-3 with the acclimatized consortium. A molecular analysis of the microbial community existing in the reactors of Run-2 and Run-3 indicated that the improvement in process performance was closely related to the selection and enrichment of specific hydrolytic–acidogenic bacteria in the reactor. A functional analysis showed that the gene copy numbers for pyruvate synthesis and fatty acid synthesis and metabolism pathways were higher in all bacterial species in Run-3 compared to in those of the other two runs, indicating improved capacity through acclimation in Run-3. The experimental results demonstrate that the hydrolysis of food waste can be enhanced through the acclimation of seed microbes from cow manure.