Lower efficiency serves as limiting factor in the enzymatic saccharification of food waste (FW) for ethanol fermentation. We investigated the effects of temperature, tetracycline; the optimization of dual enzymatic inoculation using simplex lattice design; as well as batch ethanol fermentation based on optimized FW saccharification. The moderate temperature (50 C) was proper to balance the saccharification efficiency and reducing sugar loss. The tetracycline inoculation (80 mg/L) dramatically prevented the reducing sugar loss of saccharification by inhibiting the microbial contaminants. The optimized ratio of -amylase and amyloglucosidase was 0.5:0.5, which indicated that the experimental results under the optimum conditions agreed well with the model predictions. The ethanol yield reached 0.25 g/g dry FW under optimum conditions of saccharification indicating this strategy could be promising to utilize FW.