As a new feedstock, Mespilus Germanica (M. germanica) kernel oil was used for the first time to produce biodiesel by Al 2 O 3 /CaO nanocatalyst and optimize the transesterification process operating conditions using Taguchi method. Considering Brunauer-Emmett-Teller (BET) and scanning electron microscopy (SEM) results, the applied catalyst had an adequate surface area and appropriate surface porosity for biodiesel production. Additionally, gas chromatography analysis showed that linoleic acid (42.76%) and oleic acid (40.57%) were the main constituents of the extracted oil.Based on experiments output the highest transesterification yield which was achieved in the optimal conditions (25 C, 2 h, methanol to oil ratio of 12:1, and catalyst wt of 0.5%) was 96.68%, which was an excellent yield compared to similar works.The results showed that the overall trend of production yield with temperature was downward while it saw a significant increase with time. Besides, the conversion yield registered a rise after the initial decrease with methanol to oil ratio and catalyst concentration had the opposite effect. Additionally, regeneration studies showed that the current catalyst can be used for 3 cycles with acceptable conversion yields. The physicochemical properties of the derived oil were characterized and compared with the ASTM D5671 and EN14214 standards, which were found to be highly comply with them.