A lipase from Aspergillus niger F044 was purified to homogeneity using ammonium sulfate precipitation, dialysis, DEAE-Sepharose Fast Flow anion exchange chromatography and Sephadex G-75 gel filtration chromatography. This purification protocol resulted in a 73.71-fold purification of lipase with 33.99% final yield, and the relative molecular weight of the enzyme was determined to be approximately 35～40kD using SDS-PAGE. The optimum pH and temperature for lipolytic activity of the lipase was 7.0 and 45℃, respectively. It was extremely stable at 60℃ and retained 98.70% of its original activity for 30min. The stability declined rapidly as soon as the temperature rose over 65℃. The lipase was highly stable in the pH range from 2.0 to 9.0 for 4h. Ca²⁺ and Mg²⁺ ions stimulated lipolytic activity, whereas Mn²⁺, Fe²⁺ and Zn²⁺ ions caused inhibition. The values of K_m and V_max calculated from the Lineweaver-Burk plot using pNPP as hydrolysis substrate were 7.37mmol/L and 25.91μmol/(min·mg), respectively. The N-terminal sequence of the lipase was Ser/Glu/His-Val-Ser-Thr-Ser-Thr-Leu-Asp-Glu-Leu-Gln-Leu-Phe-Ala-Gln, which is highly homogeneity with that of lipase, as reported by Torossian.