Lipase from Candida cylindracea has been purified by ammonium sulfate precipitation, sodium deoxycholate treatment, ethanol-ether precipitation and chromatography on SE-Sephadex and Sephadex G-100 columns. The purification of the enzyme was 33.4-fold with a yeild of 18.0% on the basis of activity per weight of protein. The purified enzyme was homogeneous on ultracentrifugation and electrophoresis. Optimum pH for the hydrolysis of olive oil was 7.2 by the assay method using a polyvinylalcohol-emulsified system and 5.2 by the assay method using a shaken system without a macromolecular emulsifier. Optimum temperature was 45°C. The enzyme was stable up to 15°C and in the range of pH from 2.0 to 8.5. Sodium taurocholate showed either an activating or an inhibiting effect at pH 7.0, depending on the sodium taurocholate concentration and on the assay system.