Magnetic silica chitosan microspheres (MSC) were prepared by orderly grafting SiO2 and chitosan layer on the surface of Fe3O4 nanoparticles, and after modified with epoxy groups, the microspheres were used for immobilization of naringinase. The pH, temperature, storage and operational stability of naringin hydrolysis by the immobilized naringinase were investigated. The optimal immobilization conditions were determined as follows: immobilization pH, temperature and time were 3.0, 30 ℃ and 4 h, respectively, and 57.48 U/mL of crude naringinase broth activity. As a result, the binding efficiency, activity recovery rate and specific activity of the immobilized naringinase were 31.29%, 88.92% and 409.33 U/g, respectively. Comparing with free naringinase, the immobilized naringinase used for naringin hydrolysis showed better pH stability and thermal stability, and it retained 53.36% residual activity after 8 consecutive operations and 80.97% residual activity after one month of storage.