In this study, Lipase from Aspergillus niger after extraction of medium culture was precipitated with different percentages of acetone and purified by ion exchange chromatography using SP-Sepharose HP and Q-Sepharose HP. The process of purification of the enzyme was studied by electrophoresis and the molecular weight was detected and determined by Zymography using overlying containing phenol red and Rhodamine B. The results showed that the vast majority of lipase from this strain has been precipitated by 70% saturation acetone, and leads to the 1.67 fold the purified enzyme, with special activities 32.8 U. mg-1 and efficiency of 38.5%. Using two-phase chromatography, enzyme specific activity reached 246.47 U. mg-1and 12.59-fold purification were achieved. The results of Zymography and electrophoresis indicate a lipase band with weighing about 30 kDa.

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