Abstract: The Skipjack Tuna cooking liquor was chosen as a material to produce flavor seafood sauce. With degree of hydrolysis as index, the solubilized proteins contained in flavor seafood sauce was collected and hydrolyzed by five commercial enzymes, including neutral protease, alkaline protease, papain, pepsin and pancreatin. Also response surface methodology was employed to optimize enzymatic hydrolysis technology on the base of one-factor-at-a-time experiment. Orthogonal experiment (L₉ (3⁴))was used to research the influence of yeast dosage, fermentation temperature, fermentation time on odor value. And sterilization condition was determined. The experiment results indicated that the suitable enzymatic technological conditions for Skipjack Tuna cooking liquor were papain with a dosage of 300 U/g, temperature 56℃, and hydrolysis for 4 h, the enzymatic liquid added the amount of yeast 0.75%, fermentation temperature and the reaction time were 35℃ and 60 min, the method had much deodorization efficient. The optimal sterilization was 90℃ for 30 min. The tuna sauce was developed with yellow-brown and a strong fish flavor, moreover, the content of amino acid nitrogen was 0.68 g/100 mL, can be fully developed and utilized. Thus, it is a kind of natural condiment with great development prospect.