The preparation of chitosan-coated magnetic nanoparticles (MNPs) and covalent immobilization of a-amylase for starch hydrolysis was investigated. Surface morphology, chemical composition, and structural characteristics of the MNPs were analyzed by scanning electron microscopy, energy dispersion spectrometry, and X-ray diffractometry, respectively. Surface functional groups of MNPs, chitosancoated MNPs, and a-amylase-immobilized MNPs were characterized by Fourier transform infrared spectroscopy. Response surface methodology based on three levels was implemented to optimize three immobilization conditions and a regression model was developed. a-Amylase-immobilized MNPs provided better stability towards pH and temperature. The prepared thermostable nanobiocatalyst is well-suited for industrial processes involving starch hydrolysis.a-Amylase, chitosan, ferric chloride, ferrous sulfate, ammonium hydroxide, and solvents were purchased from Hi-media, www.cet-journal.com