This paper describes a new support that permits to efficient immobilization of L-asparaginase (L-ASNase). For this purpose, Fe3O4 magnetic nanoparticles were synthesized and coated by MCM-41. 3-chloropropyltrimethoxysilane (CPTMS) was used as a surface modifying agent for covalent immobilization of L-ASNase on the magnetic nanoparticles. The chemical structure; thermal, morphological, and magnetic properties; chemical composition; and zeta potential value of Fe3O4@MCM-41-Cl were characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), differential thermal analysis (DTA), differential scanning calorimetry (DSC), vibrating sample magnetometer (VSM), scanning electron microscope (SEM), energy dispersive X-ray (EDX), X-ray diffraction patterns (XRD), and zeta-potential measurement. The immobilization efficiency onto Fe3O4@MCM-41-Cl was detected as 63%. The reusability, storage, pH, and thermal stabilities of the immobilized L-ASNase were investigated and compared to that of soluble one. The immobilized enzyme maintained 42.2% of its original activity after 18cycles of reuse. Furthermore, it was more stable towards pH and temperature compared with soluble enzyme. The Michaelis-Menten kinetic properties of immobilized L-ASNase showed a lower V-max and a similar K-m compared to soluble L-ASNase. Immobilized enzyme had around 47 and 32.5% residual activity upon storage a period of 28days at 4 and 25 degrees C, respectively. In conclusion, the Fe3O4@MCM-41-Cl@L-ASNase core-shell nanoparticles could successfully be used in industrial and medical applications.