In this study, transesterification and esterification were investigated in batch and continuous process using immobilized Candida rugosa and Rhizopus oryzae lipases. In the case of batch process, stepwise reaction method was investigated to prevent the lipase deactivation. Reaction conditions were as follows: temperature, 45 A degrees C; agitation speed, 250 rpm; enzyme concentration, 20%; and water contents 10%. And then, conversion yield was 98.33% at 4 h. In the case of continuous process, circulation and long-term continuous system were investigated for development of efficient mass transfer system. Optimal reaction conditions were as follows: temperature, 45 A degrees C; flow rate, 0.8 mL/min; and water contents, 10%. And then, conversion yield of biodiesel was 97.98% at 3 h. Especially, the maximum conversion yield using a mixture of immobilized lipases exceeded over 90% for 108 h in long-term continuous system under optimal reaction conditions (45 A degrees C; flow rate, 0.8 mL/min; and water contents, 10%). These results should help in determining the best method for the biodiesel production and improving the design and operation of large scale by enzymatic systems.