The transesterification of canola oil with methanol was studied in a heterogeneous system, using gamma-Al2O3 supported alkaline catalysts. All the catalysts were prepared by incipient-wetness impregnation of an aqueous solution of alkaline compounds on gamma-Al2O3 support. The effects of alkaline compound, methanol/canola oil molar ratio, reaction temperature, catalyst amount, and reaction time in biodiesel production were investigated. The catalyst with KOH loaded on gamma-Al2O3 gave the highest basicity and the best catalytic activity for this reaction. The highest FAME yield obtained was 89.40%.