The invention discloses anti-senility co-immobilized enzyme which includes catalase and glucose oxidase according to the activity ratio being 8:1-15:1. A preparation method of the anti-senility co-immobilized enzyme includes the steps of: a) according to the activity ratio, dissolving the catalase and the glucose oxidase in a phosphate buffer solution to prepare a mixed enzyme solution of the catalase and the glucose oxidase, and adding chitosan microspheres to the mixed enzyme solution to obtain a mixed liquid; b) stirring and adsorbing the mixed liquid; c) adding a glutaraldehyde solution according to mass percentage ratio to perform a crosslinking reaction; d) allowing the liquid to stand to balance the liquid to room temperature; and e) adding [beta]-D-glucose, electromagnetically stirring the liquid and allowing the liquid to stand for later use. Through synergistic effects of the catalase and the glucose oxidase, hydrogen peroxide, which is generated by reducing oxygen with bivalent electrons under external conditions, such as ultraviolet and the like, on skin surface, can be removed, thus alleviating senility.