This study aimed to determine the optimal conditions for bioethanol production from pineapple peels using response surface methodology at high temperatures. Evaluation of factors that affected on ethanol production at 40^oC was carried out with 7 factors (urea, NH₄NO₃, MgSO₄.7H₂O, KH₂PO₄, yeast extract, pH, and yeast density) using the statistical model of Plackett-Burman Design. Optimizations of conditions for ethanol production were conducted with the response surface methodology using Central Composite Design. As a result, the optimal conditions of ethanol production from pineapple peel hydrolysate by S. cerevisiae YVN8 at 40^oC were identified as followed: 0.30 g/L of NH₄NO₃, 0.64 g/L of yeast extract, and pH 4.41. In the optimal conditions, the ethanol concentration was attained at 33.3 g/L.