Aims: Bioethanol is an environmental friendly energy source with a lot of great prospective and become an alternative to fossil fuels .Oil palm frond juice (OPFJ) is a potential sources of sugars for bioethanol production. The present study aimed to optimize bioethanol production. Methodology and results: Bioethanol fermentation was carried out by Saccharomyces cerevisiae HC10 using OPFJ as substrate in bioreactor with 1.5 L working volume. Growth profile was performed for 42 h with sampling every 3 h interval. Effect of agitation speed (rpm) and volume of OPFJ were screened to select significant factor for high production of bioethanol. Agitation speed at 175 rpm and volume of oil OPFJ; 40% gave 5.25 g/L and 4.52 g/L of ethanol and biomass concentration, respectively. These parameters were further investigated via central composite design (CCD) of Response Surface Methodology (RSM) to maximize bioethanol production. The suggested optimum conditions for bioethanol production were agitation speed at 152 rpm and volume of OPFJ at 39.71% in which giving ethanol concentration of 4.79 g/L. Growth profile after optimization indicated that the highest ethanol concentration (5.75%) was achieved after 15 h of fermentation. Kinetic studies indicated that ethanol yield coefficient (Yp/s) due to consumption of OPFJ and productivity of ethanol was 3.5 fold and 25% increased compared to before optimization, respectively. While, in term of ethanol yield about 9% increased was observed. Conclusion, Significance and Impact of study: This showed that OPFJ can be an alternative new feedstock for bioethanol production using S. cerevisiae HC10.