Abstract: To optimize the ultrasinic- assisted enzymatic extraction of polysaccharides from Cantharellus cibarius, on the base of the single factor experiment results, enzymatic hydrolysis p H, solid- liquid ratio, ultrasonic power and enzymatic hydrolysis time were identified as main factors that influence polysaccharides yield with the Plackett-Burman experiment design firstly.Then the Box- Behnken experiment design and response surface analysis were adopted to optimize the extraction conditions with polysaccharides yield as the response value. The optimized technological parameters were as follows: p H5.5, solid- liquid ratio 1 ∶ 46 ( g / m L) , ultrasonic power 210 W and enzymatic hydrolysis time 68 min. Under these conditions, the theoretical and actual crude polysaccharides yield were 12.08% and 11.91%, respectively, with a relative error of 1.41%, which indicated that the combination of Plackett- Burman design and response surface analysis can well optimize the extraction technology of polysaccharides from Cantharellus cibarius with the ultrasinic- assisted enzymatic extraction method and it can provide certain technical support for the widely used of Cantharellus cibarius.