Lipase has attracted a lot of attention in recent years because of its diverse biotechnological applications, like biopolymer synthesis, biodiesel production, treatment of fat-containing waste effluents, enantiopure synthesis of pharmaceuticals and nutraceutical agents, have been established successfully. Lipase occurs widely in nature, but plant lipase, is significant because of its unique substrate selectivity/specificity. However new lipases with properties amenable for application in specific industrial process are being sought by the researchers. In this work, a penicillium strain was isolated that showed lipase production from endosperm tissues of decaying seeds of Cucummeropsis mannii obtained from the regular Ogige market in Nsukka metropolis, Enugu state. The optimization studies resulted to an increase in enzyme activity: the strain produces lipase in an inexpensive inorganic medium at highest when ammonium sulphate is used as a source of nitrogen when compared with lipase activity on other sources of inorganic nitrogen. Lipase production known to be induced when lipids such as vegetable oils (triacyiglycerides) are included in the medium as carbon sources. The potential to induce lipase production is dependent on the type of oil used. The inclusion of olive oil in medium showed that lipase extract from Cucumeropsis mannii had its highest enzyme activity when it is used as a carbon source when compared to other sources of carbon like palm kernel oil, groundnut oil, soyabean oil and sunflower oil. Further studies on the lipase activity from Cucumeropsis mannii also showed that the lipase produced from it was highest at pH 8.0, which meant that the lipase is best produced at an optimum medium pH of 8.0.