Abstract: The technology for preparation of anti-hypertensive peptides from Alaska cod skins with optimum enzyme was optimized by using the index of ACE inhibitory rate in vitro. In this study,the optimal enzymatic hydrolysis process of anti-hypertensive peptides from Alaskan cod was determined by single-factor and orthogonal experiments,and the amino acid content and molecular weight distribution range of anti-hypertensive peptides were investigated. The results showed that the trypsin had the best hydrolyzing capacity,the ACE inhibition rate reached a constant value of 68.48%. And the optimum hydrolysis conditions were as follows:Hydrolysis temperature 40℃,the ratio of enzyme to substrate 3%,the ratio of solid to liquid 1:7,pH7,hydrolysis time 2 h. Under these conditions,the ACE inhibition rate was 79.62%,content was 83%,the relative molecular weight below 500 Da was 70.74%,respectively. The optimized extraction process of anti-hypertensive peptides from Alaskan cod was feasible and reasonable. The results of this study could provide a reference for further investigation of food-derived anti-hypertensive peptides.