The source of catalyst and feedstock used for transesterification of oil play a significant role in cost, and starvation of consumers of oil especially for edible oil sources. This investigation aimed at the transesterification of castor oil with the aid of calcined anthill as the catalyst support. Box Behnken Design (BBD) of the Response Surface methodology was employed to study the relationship among the variable of production such as time (1 – 3 h), temperature (40 – 60oC), catalyst loading (1 – 9 g), methanol volume (10 – 60 ml) and agitation rate (100 – 400 rpm) on yield of biodiesel produced. The synthesized catalyst was characterized using X-Ray Fluorescence (XRF) while biodiesel produced was characterized through determination of physicochemical characterized and function group applicable to biodiesel. The calcined anthill was found to contain some mixed basic oxides. The physicochemical properties of produced biodiesel are within the American Standard Test Methods (ASTM) range (except density that is higher), and the functional groups found in biodiesel are typical of vibrations presence in biodiesel. The optimum condition for a volume of 11.73 ml, agitation of 166.458 rpm, reaction time of 2.34375 hrs, temperature of 59.4552oC, catalyst loading of 1.14 g, and an optimal yield of 90.59 %.