The present communication deals with the production of lipase from Penicillium sp. using waste oils and palm cactus (Nopalea cochenillifera) especially as nutrient source of low cost. Two different waste oils were tested: waste frying oil from an industrial kitchen and waste lubricating oil (WLO) from a gas station. Using Doehlert experimental design and response surface methodology, the optimum conditions for lipase production were 96h fermentation, WLO as the inductor, with specific activity of 0.22UAmg(-1). The enzyme was able to remain with more than 58% of its original activity until 30min at 60 degrees C. The kinetic constants were K-m=9.93mM and V-max=2.58 UA min(-1) using p-nitrophenyl palmitate (p-NPP) as substrate. Results showed that Penicillium sp. was able to produce lipase from waste oils using N. cochenillifera, thus having biotechnological potential in waste oil biotransformation.