Biodiesel was produced from an oil fraction separated from food waste leachate, using a batch reactor system, to evaluate its potential as a renewable energy source for the first time. In order to reduce the free fatty acid content of the leachate oil fraction, three different acid catalysts, H2SO4, zeolite and Amberlyst-15, were used in the pretreatment process. The residual oil fraction after pretreatment was further treated over KOH as a base catalyst to produce a fatty acid methyl ester (FAME). The FAME content in the product was analyzed using GC/MS. A large number of different FAMEs were detected, which is characteristic of the food-waste-derived raw material used in this study. The FAME content of the product was highest when pretreated over Amberlyst-15, followed by trans-esterification. Additional refining and process optimization would be expected to further increase the FAME content. Therefore, it was concluded that the production of biodiesel from the oil fraction separated from food waste leachate is promising.