A hydrophobic group/comonomer N-methacryloyl-(l)-alanine (MAAL) containing 2-hydroxyethyl methacrylate (HEMA)-based magnetic hydrophobic nanoparticles was synthesized by using a microemulsion polymerization technique for immobilization of amyloglucosidase (AMG). The magnetic hydrophobic nanoparticles with an average diameter of 79 nm were characterized with Fourier transform infrared spectroscopy, transmission electron microscopy, zeta-size analysis, and electron spin resonance spectroscopy. Hereby, the effecting factors such as initial concentration of AMG, pH, and temperature on the immobilization were investigated. After determining the optimum immobilization conditions, the kinetic constants (K-M and V-max) and properties of free and immobilized AMG were investigated in batch studies. The maximum AMG adsorption capacity of the magnetic hydrophobic nanoparticles was 294.42 mg/g at pH 3.0. The optimum pH and temperature for free and immobilized AMG were found to be pH 5.0 and 60 degrees C. The K-M value for immobilized AMG was higher than that of the free enzyme, whereas the Vmax value was lower for the immobilized AMG preparations.