The invention discloses a method for preparing water-insoluble beta-1, 3/1, 6-qlucan. The method has the steps as follow: 1)adding lysozyme into yeast cell wall suspending liquid for enzymolysis, centrifugation and collection at a temperature between 35 and 40 DEG C; 2) putting the product achieved from step 1) into NaOH solution and stirring for centrifugation and collection at a temperature between 35 and 4 DEG C; 3) depositing the product achieved from step 2) into acetic acid solution having concentration expressed in percentage by volume of 2-8 percent and stirring for centrifugation and collection; 4) stirring and washing the deposition after acid treatment and centrifugation of the step 3)by absolute ethyl alcohol and acetone and ether extract sequentially, and laying aside the treated ether extract for 12 to 24 hours, and drying in the centrifugation and collection way to achieve water-insoluble beta-1, 3/1, 6-qlucan. The invention is simple in control, and the beta-1,3-qlucan and the 1,6-qlucan prepared by the method have high purity and yield, and has the protein content and the fat content less than 1 percent.