Biodiesel (fatty acid methyl esters, FAME) is an important biofuel for the future. It is produced by transesterification of triglycerides, in vegetable oils and animal fats. In the first-generation processes, alkaline-based catalysis was mostly employed. It has been established for the conversion of vegetable oils with relatively low free fatty acid content [1,2]. In second generation processes, enzymatic reaction was commonly applied, using a lipase (EC 22.214.171.124) [3-5]. Enzymatic method is anticipated as an environmental friendly processes and lower consumption of thermal energy. Several scientific reports have discussed the use of immobilized lipases. For establish lower cost in application, direct use of liquid soluble lipase is favorable for commercial processing. Several recent publications attest to the clear advantages of liquid soluble in application. Callera Trans L™ contained Thermomyces lanuginosus (Novozymes A/S, Denmark) has been used often in investigations. For an effective commercial operation, physicochemical properties of aqueous-oil interfacial area are important subject since lipases are activated at the liquid-liquid interface.