This study discribes performance of DEFC (Direct Ethanol Fuel Cell) utilized bio-ethanol based on fruit wastes. To produce the bio-ethanol, fruit wastes were treated at temperature 120℃ and 90minutes in acid pre-treatment. After pre-treatment was done, alcohol fermentation process was running. Initial alcohol concentration was 5%. Using the multi coloumn distillation system, more than 95% ethanol was distilled and each component of bio-ethanol was analyzed. In DEFC performance test, it was revealed that cell performance was much higher than that of ethanol. Comparing ethanol with mixed fuel (bio-ethanol (10%) + ethanol (90%)), the performance of ethanol was higher than that of mixed fuel. Even though the bio-ethanol from the fruit wastes is corresponded with transport ethanol standards, it thought that organic matter in bio-ethanol could be negative effect on fuel cell.