Abstract: The yeast extract was prepared using Candida utilis yeast as raw materials. The key factors affected autolysis of yeast was discussed with indicators of yield of amino nitrogen and yeast extract. And orthogonal experiment design showed that the concentration of yeast 15% ( w / w) , 75 MPa homogeneous pressure 2 times, p H6.5, 50℃, addition of 1.5% flavour enzyme and autolyzed 48 h were the best optimizing autolytic environment.Under these conditions, the yield of amino state nitrogen and yield of yeast extract were 650.6mg /100 m L and50.1%, respectively.And the yeast extract owns Maotai- flavor and slightly ester aromat, with taste rich and fresh sweet.