Abstract The objective of this study was to investigate the effects of particle size reduction and solubilization on biogas production from food waste (FW). To clarify the effects of volatile fatty acids (VFAs) in the digestion process, the relationship between particle size and VFA accumulation was investigated in detail. For this purpose, substrates of various particle sizes were prepared by bead milling to support hydrolysis. Batch anaerobic digestion experiments were carried out using these pretreated substrates at mesophilic temperature for a period of 16 days. The results of pretreatment showed that the mean particle size (MPS) of substrates ground with a bead mill decreased from 0.843 to 0.391 mm, and solubilization accounted for approximately 40% of the total chemical oxygen demand (total COD) for grinding pretreatment by bead milling. Anaerobic digestion batch experiments revealed that MPS reduced by bead milling at 1000 rpm improved methane yield by 28% compared with disposer treatment. Moreover, this may have increased microbial degradation during the VFA production process with increasing total number of revolutions (operation time × revolutions per minute). However, excessive reduction of the particle size of the substrate resulted in VFA accumulation, decreased methane production, and decreased solubilization in the anaerobic digestion process. These results suggest that optimized reduction of the particle size of the substrate in conjunction with optimized microbial growth could improve the methane yield in anaerobic digestion processes.