Enzymatic production of biodiesel from palm oil using partially purified lipase from the hepatopancreas of Pacific white shrimp (Litopenaeus vannamei) as a low cost catalyst was studied. In this optimization study, the effects of reaction time, enzyme concentration, methanol/oil molar ratio, water content and reaction temperature on the biodiesel yield were considered. The best conditions for biodiesel preparation were: 70 kUnit lipase, 4:1 methanol to oil molar ratio, 3% water, 45 degrees C reaction temperature and a reaction time of 16 h. Under these optimal conditions, the maximum biodiesel yield reached 97.01%. The attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and thin layer chromatography (TLC) were used to ensure the conversion of palm oil into biodiesel. The biodiesel properties were within the recommended biodiesel standards as prescribed by EN 14214 and ASTM D 6751. Therefore, the lipase from the hepatopancreas of Pacific white shrimp is a potential biocatalyst for the biodiesel industry. (C) 2016 Elsevier Ltd. All rights reserved.