Abstract: Aqueous enzymatic extraction of avocado oil with pineapple juice in low-temperature was investigated.Protease activity of pineapple juice at different solid contents was measured by ultraviolet spectrophotometer.The effects of different factors such as pH, reaction time and the like on the extraction rate of avocado oil were measured through single factor test and then the extraction conditions were optimized by orthogonal test. The results showed when the pineapple juice with a solid content of 2° Brix was added into the avocado pulp, adjusting the ratio of material to liquid to 1∶7g/mL;then the enzymatic reactions within the above mixed solution were undertaken for 4h at the temperature of 45℃ and natural pH value, followed by an optimum oil extraction yield of 75.38%. This method for recovering oil from fresh avocado pulp with pineapple juice was a significant improvement in both oil yield and quality compared with the traditional hot process.