Abstract: Emulsion was formed during the process of enzyme-assisted aqueous extraction of perilla frutescens seeds oil.It is stabilized by oil, proteins, phospholipids and carbohydrates, which compose a state emulsified system, as it is rich in oil, therefore, how to demulsify effectively the emulsion, which is the key to increase the oil yield of enzyme-assisted aqueous extraction processing (EAEP) of oil.Emulsion formed in the emulsion from enzyme-assisted aqueous extraction processing (EAEP) of Perilla Frutescens Seeds oil was taken as study object, and econdary hydrolysis de-emulsification was adopted to demulsify the emulsion, the effect of hydrolysis time, hydrolysis temperature, hydrolysis pH and enzyme dosage on oil recovery yield.Response surface methodology (RSM) was applied to optimize the demulsification technology.Optimal hydrolysis conditions were Protex 6L, hydrolysis time of 1.6h, hydrolysis temperature of 62.6℃, hydrolysis pH of 9.4 and enzyme dosage of 1.9%.Under these optimal hydrolysis conditions, the maximum oil recovery yield was up to 85.52%.For hydrolysis de-emulsification, it was a good method that feasible, safe operation, and potential for industrial application.