Abstract: The effect of enzymes( neutral protease,papain,pepsin,trypsin and flavourzyme) on extraction rate of Andrias davidianus oil were studied by the single factor experiment,and papain was selected to be the enzyme for the experiment. Central composite design( CCD) were used to optimize the condition of aqueous enzymatic process. The optimal conditions were as follows: solid- to- liquid ration of 1 ∶ 1( w / w),papain added dosage of6800 U / g,p H5.8,hydrolysis temperature of 50 ℃,and hydrolysis time of 130 min,and under such condition the extraction rate was 93.52%.Chemical and sensory evaluation results showed that the crude Andrias davidianus oil extracted by aqueous enzymatic conformed to the secondary standards crude fish oil of SC / T 3502-2000.