Abstract: Central composite design(CCD)-response surface(RSM) was used to optimize the aqueous enzymatic extraction process of Perilla frutescens seed oil. On the basis of single factor experiment, we adopted the central composite design method to study the effects of enzymatic species, enzymatic hydrolysis temperature, pH, liquid-solid ratio, enzyme dosage and time interaction on the extraction rate of Perilla oil. The fitting equation was significant, the optimum conditions for the extraction of Perilla oil were as follows:pH is 9.5 in alkaline protease, the solid-liquid ratio(g) was 9.97:1, the amount of enzyme added was 2.75%, the temperature was 56.1℃, and the time was 5.25 h. Under this condition, the extraction rate of Perilla oil could reach 37.65%, which was very close to the theoretical value of 38.3%. The established model was true and reliable, and the optimum extraction process of Perilla oil was determined. The Perilla seed oil contains palmitic acid, stearic acid, oleic acid, linoleic acid and alpha-linolenic acid by gas chromatography, the relative content of alpha-linolenic acid in Perilla oil extracted by aqueous enzymatic method was 67.9%, and the physical and chemical indexes of relative solvent method and cold pressing method were the best.