Abstract: Used squid liver as the raw material, adopted the AS.1389 neutral protease to extract fish oil, chose extraction rate as index, the best enzymatic technology was confirmed by single factors and surface response tests, and the fish oil extraction ratio, oil quality and fatty acid composition were compared which were extracted by enzymatic and water. The result showed that the optimum condition of enzymatic technology was follows:liquid ratio was 1:1, pH was 6, the quantity of enzyme was 1 340 U/g, temperature was 55℃ and hour was 4 h; Compared with water extraction, the enzymatic oil were higher extraction rate, lower peroxide value, better oil quality.