Biodiesel production from Ceiba pentandra oil using immobilized Candida antarctica lipase (Novozym 435) was studied. A drastic change from 20.6% to 78.0% yield for 9:1 methanol to oil molar ratio indicated the significance of methanol stepwise addition method. Tests were done on the effect of enzyme pretreatments to biodiesel yield productivity which resulted in increase of yield when using t-butanol but decreased yield when using sodium chloride solution. The reusability of enzyme was measured at 63.7% relative yield after three batches. Biodiesel production from C. pentandra oil using biocatalyst is feasible and can be improved for industrialization.