Lipases are one of the industrially important biocatalyst whose demand ranks third among all the other enzymes. Because of their ability to carry out diverse range of reactions, remain active even in water restricted environment and act at interfaces, they find applications in varied fields. Purification and characterization of this enzyme is essential for its application in a wide spectrum of industrial processes. The present study deals with purification of lipase from Rhizopus oryzae NRRL 3562 in minimum steps which led to a high yield and fold increase. Lipase with specific activity of 450 IU/mg gave a single band on both native and SDS PAGE, showing purification to homogeneity. It was found to be a low molecular weight lipase of 14.45 kDa and had optimum temperature range and pH of 30-40○C and 9, respectively. The purified lipase showed specificity towards long chain (C16-18) p-Nitrophenyl esters. Stability at
elevated temperatures, wide pH range and in presence of solvents makes this small lipase a potent candidate for transesterification reactions.