Technical Abstract: Lipases are industrially and useful versatile enzymes that catalyze numerous different reactions including hydrolysis of triglycerides, transesterification, and chiral synthesis of esters under natural conditions. Although lipases from various sources have been widely used in industrial applications, such as in food, chemical, pharmaceutical and detergent industries, there are still substantial current interests in developing new microbial lipases, specifically those functioning in abnormal conditions. We screened 17 lipase-producing yeast strains, which were pre-screened for substrate specificity of lipase from more than 500 yeast strains from the Agricultural Research Service Culture Collection (Peoria, IL, USA), and selected Yarrowia lipolytica NRRL Y-2178 as the best lipase producer. This report presents new findings and optimal production of a novel extracellular alkaline lipase from Y. lipolytica NRRL Y-2178. Optimal culture conditions for lipase production by Y. lipolytica NRRL Y-2178 were 72 hours incubation time, 27.5', and pH 9.0. Glycerol and glucose were efficiently used as carbon sources and a combination of yeast extract and peptone was a good nitrogen source for lipase production by Y. lipolytica NRRL Y-2178. These results suggest that Y. lipolytica NRRL Y-2178 has industrial potential as a new alkaline lipase producer.