This experiment focused on biodiesel production using Indian oil sardine fish as a new low-cost feedstock. Bligh and Dyer's method was followed for oil extraction from Indian oil sardine fishes. Biodiesel was produced by KOH catalyzed transesterification reaction with a reaction temperature of 150 degrees C. Optimization of the process parameters influencing the biodiesel production was performed using response surface methodology (RSM). BoxBehnken experimental design was used for the statistical analysis. The effect of methanol vol%, KOH wt% and reaction time over the reaction were studied. Chemical characterization of the biodiesel produced was carried out by Gas Chromatography-Mass Spectroscopy (GCMS) analysis. Physicochemical properties of the biodiesel were characterized according to ASTM 6751 standard. The optimum process conditions for the biodiesel yield of 96.57% were found to be 20 vol% methanol, 1.25 wt% KOH and 25 min reaction time. The 150 degrees C reaction temperature used in this process enhances the transesterification reaction by reducing the reaction time from the normally observed 60-120 min to 25 min with a high yield of biodiesel.