Abstract: In order to extractβ- D- Glucosidase from Oenococcus oeni 31MBR effectively, response surface methodology was used to optimize ultrasonic conditions on the basis of single factor test.The results showed that optimum conditions were: ultrasonic time 20min, ultrasonic power 130W, and the cell concentration about OD600=2.0. Under such conditions, totalβ- D- glucosidase activity from 31MBR was about 13.89U, which was higher than repeated freezing and melting and glass- bead. β- D- Glucosidase could be extracted from Oenococcus oeni effectively by ultrasonic, and it was feasible to optimize ultrasonic extraction conditions using response surface methodology.