Abstract: Using the viscera of black carp as raw material,fish oil was extracted by microwave-assisted protease. The effects of enzymatic type,amount of enzymatic addition,enzymatic hydrolysis time,enzymatic hydrolysis temperature,microwave power and microwave treatment time on the yield of fish oil were studied. Regression equation between influencing factors and fish oil extraction rate was established by using Box-Behnken response surface design method. The results showed that neutral protease was the best hydrolytic enzyme and the optimized extraction conditions for fish oil were as follows:neutral protease was the best hydrolase,micro wave power 400 W,enzyme dosage 2%,enzymatic hydrolysis time 2.5 h,enzymatic hydrolysis temperature 45℃,and microwave treatment time 15 min. Under these conditions,the fish oil yield was 26.26%±0.13%.Qualitative and quantitative analysis of fatty acids composition were done in extracted visceral fish oil of black carp by gas chromatography(GC). The analysis results showed that there were three kinds of saturated fatty acids,accounting for 22.39%,mainly palmitic acid and stearic acid. There were three kinds of monounsaturated fatty acids,accounting for 40.42%,mainly oleic acid and palmitoleic acid. There were eight kinds of polyunsaturated fatty acids,accounting for 37.19%,mainly linoleic acid.