Abstract: The cell wall of wine yeast was broken by high pressure homogenization and freezing-thawing methods collaboratively. Combined with enzymatic hydrolysis of compound protease and lipase,the effect of multiple cell-wall-broken technologies on purity of β-Glucan was studied. On the basis of single factor experiment,the Box-Behnken design was adopted. The concentration of yeast,homogeneous time and freezing-thawing water content were factors,and the purity of β-Glucan was response value,the extraction process of β-Glucan from waste wine yeast was optimized. The results showed that the optimal extraction parameters were obtained as follows:homogeneous pressure 70 MPa,concentration of yeast 13%,homogeneous time 34 min and freezingthawing water content 25%,while extraction purity of β-Glucan content was 91.69% and yield was 13.23%under these conditions. The process provided a reference basis for the development and utilization of Glucan from yeast.