In this study, a new CaO/CuFe2O4 nanocatalyst was first synthesized and its structural and morphological properties were evaluated by using various analyzes such as SEM, FTIR, XRD, TGA, EDX and TEM. Then, bio-diesel was prepared by using chicken fat in the presence of CaO/CuFe2O4 nanocatalyst during transesterification process. To determine the highest yield of biodiesel production, the effect of different parameters such as methanol to oil ratio (4: 1-24: 1), reaction time (0.5-6 h), reaction temperature (50-80 degrees C), and catalyst content (0.5-7%) was studied accordingly. The result indicated that the highest biodiesel yield was obtained at the methanol to oil ratio of 15: 1, the reaction time of 4 h, the reaction temperature of 70 degrees C and catalyst content of 3% which was 94.52%. Subsequently, the biodiesel produced in optimum conditions was mixed with gasoline in various ratios (B100, B80, B60, B40, B30, B20 and B10). Finally, the physical and chemical properties of synthesis fuels such as density, flash point, kinematic viscosity, pour and cloud points were investigated based on the international standards of ASTM D 6751 and ASTM D 14214.