Abstract:The abalone viscera was used as raw materials，which was extraction of β-glucosidase by ultrasonic assisted，and the different ultrasonic power，ultrasonic time and extraction time of a certain amount of the pretreatment liquid product in the influence of enzyme activity was discussed.The optimum process conditions were obtained by single factor test and orthogonal test.On this basis，polyethylene glycol（PEG）/（NH4）2SO4 aqueous two-phase system was applied to extract β-glucosidase from crude enzyme fluid and optimize the twophase system.The experimental results showed that ultrasonic power had a significant effect on the extraction，ultrasonic time had no significant influence，and the extraction time after ultrasonic could be neglected，so when the pretreatment liquid was 10 mL，ultrasonic power was 180 W，ultrasonic time was 20 min，extraction time after ultrasonic of 1 h were the optimal process conditions.The optimal two-phase system was the mass fraction of PEG1000 at 10%，and the mass fraction of（NH4）2SO4at 16%，pH at nature（pH≈5）.