BACKGROUND: Supercritical carbon dioxide (SCCO2) has been expected to be a suitable solvent for industrial production of biochemical components using enzymatic processing. This paper presents advantages of SCCO2 for lipid hydrolysis by immobilized lipase not only for reaction rate but also for repeated use. Four lipase sources (Candida cylindracea, Candida rugosa, Rhizopus arrhizus, and wheat germ) were employed to examine rapid initiation and higher production of fatty acid. They were immobilized using a porous polypropylene particle carrier (Accurel MP100). RESULTS: The amount of lipase adsorbed on polypropylene particles pretreated with ethanol was logarithmically correlated with the molecular hydrophobicity of the lipase. The immobilized yield of lipase based on the adsorbed amount exceeded 98% for every tested source. In particular, immobilized R. arrhizus lipase exhibited rapid initiation and higher production of oleic acid. In five repeated uses of immobilized lipase in SCCO2, the rapid initiation was highly reproducible and a satisfactory reaction yield was successfully obtained. CONCLUSION: The excellent advantages of SCCO2 for lipid hydrolysis by immobilized lipase on polypropylene particle carriers were demonstrated, showing higher reaction rate and reproducible repeated use, promising findings expected to be applied to industrial process development. (C) 2015 Society of Chemical Industry