Abstract: The comparison of extraction of pork endogenous lipase with three methods of PEG2000/ (NH 4) 2 SO4 aqueous two-phase system extraction and reverse micelles extraction and combination of aqueous two-phase extraction and chitosan precipitation in this paper.Further the extraction conditions for each method were optimized by the orthogonal experiment.The optimized condition was gained:mass fraction of PEG2000 25%, mass fraction of (NH 4) 2 SO412%, pH6.5, Chitosan concentration of 2.5mg/mL.Under the condition of aqueous two phaseextraction and chitosan precipitation phase combination, lipase extraction rate was 97.83%, the maximum distribution coefficient 5.209, multiple purification was 7.74.