Biodiesel synthesis from non-edible vegetable oil via catalytic transesterification is one of the effective ways to replace petroleum-based fuels in the area of renewable energy development and is beneficial to environmental security. Therefore, this research investigates the optimization of process parameters (temperature, methanol to oil ratio, and NaOH catalyst dose) for the conversion of biodiesel from non-edible desert date (Balanites Aegyptiaca) seed kernel oil using the Box-Behnken experimental design of response surface methodology statistical analysis. Accordingly, the optimum values of reaction conditions, namely, a temperature of 60.5 °C, methanol to oil ratio of 6.7:1, and catalyst dose of 0.79 %wt, yielded 93.16% biodiesel. Fourier transform infrared spectroscopy analysis confirmed the cracking of a single glycerol backbone from the triglycerides and the substitution by methoxyl in the presence of a NaOH catalyst. The physicochemical properties of the biodiesel were investigated and compared with standards in terms of its density, viscosity, higher heating value, acid value, saponification value, cetane number, cloud point, pour point, and flash point, and the values are within the recommended standard limits of American Standard for Testing Material (ASTM D6751) and European Committee for Standardization (EN14214). Thus, the results revealed that homogeneous base catalysis of non-edible oil under optimum reaction conditions provides high yield of biodiesel.