The invention provides a method for preparing mixed immobilized glucose oxidase (GOD)/catalase (CAT) microspheres and belongs to the technical field of immobilized enzyme. The method comprises the following steps: on the basis of using chitosan-arginine anionic microspheres as carrier, adding a mixed solution of GOD and CAT and glutaraldehyde, performing crosslinking reaction, filtering, washing,and performing freeze drying to prepare mixed immobilized GOD/CAT microspheres. The method has the characteristics of mild reaction conditions, higher recovery rate of enzyme activity, long half-lifeof immobilized enzyme, simplicity in operation, low cost, cleanness, safety, and the like. The product prepared by the method can be widely used in the industries of medicines, food, water treatment,cement and the like. The product can also be used as a glucose-removing reagent, an anti-oxidation reagent, a bactericide, a glucose quantitative analysis reagent and the like.