Abstract:Objective To improve the immobilization technology of bromelain which has poor stability to enhance its utilization, stability, and storage performance.Methods The preparation process of magnetic chitosan/sodium alginate immobilized bromelain (Fe3O4-SA/CS-Br) is optimized by the response surface methodology. The Fe3O4-SA/CS-Br is characterized and its Km value, operational stability, thermal stability, storage stability, and application effectiveness are determined.Results The optimal preparation conditions for Fe3O4-SA/CS-Br are as follows: mchitosan∶msodium alginate ratio of 1∶4, Fe3O4 mass concentration of 10 mg/mL, CaCl2 mass concentration of 32 mg/mL, immobilization time of 106 min, pH value of 7.0, and enzyme mass concentration of 1.0 mg/mL. According to the results from scanning electron microscopy, infrared spectrum, and X-ray diffraction spectrum, the main functional groups of bromelain are successfully immobilized on Fe3O4-chitosan/sodium alginate (Fe3O4-SA/CS) microspheres. The affinity of bromelain for the substrate is enhanced after immobilization.Conclusion Fe3O4-SA/CS-Br can effectively enhance the stability of bromelain.