The present work demonstrated the enzymatic transesterification of the canola oil with methanol under the influence of ultrasound using commercially available lipase from Candida rugosa as a catalyst. The results clearly shed light on the importance of ultrasound in biodiesel synthesis. Although the reported ultrasonic effects vary widely, the major benefit observed from ultrasonication is the much shorter reaction time. Sonication decreases the reaction time to ∼1.5 h with the help of ultrasonic horn having probe diameter 3.5 cm as compared to 22–24 h without ultrasonic treatment. The results show that maximum yield can be obtained at power density of 2 W/cm3 using sonication horn. Moreover, plate transducers can also provide the same yield but at higher frequencies one need to work at higher sonication powers to get same amount of conversions. Ultrasonication is a promising and efficient technique for biofuel synthesis but the sonochemistry and the design of sonochemical reactors is far from being completely understood. Thus, continued efforts are required to explore this novel technical area.