alpha-Galactosidases hold immense potential due to their biotechnological applications in various industrial and functional food sectors. In the present study, soluble and covalently cross-linked aggregated forms of a low molecular weight, thermo-labile alpha-galactosidase fromVigna mungo(VM-alpha Gal) seeds were immobilized onto chitosan-coated magnetic nanoparticles for improved stability and repeated usage by magnetic separation. Parameters like precipitants (type, amount, and ratio), glutaraldehyde concentration, and enzyme load were optimized for the preparation of chitosan-coated magnetic nanocomposites of cross-linked VM-alpha Gal (VM-alpha Gal-MC) and VM-alpha Gal (VM-alpha Gal-M) resulted in 100% immobilization efficiency. Size and morphology of VM-alpha Gal-M were studied through dynamic light scattering (DLS) and scanning electron microscopy (SEM), while Fourier transform infrared spectroscopy (FTIR) was used to study the chemical composition of VM-alpha Gal-MC and VM-alpha Gal-M. VM-alpha Gal-MC and VM-alpha Gal-M were found more active in a broad range of pH (3-8) and displayed optimal temperatures up to 25 degrees C higher than VM-alpha Gal. Addition of non-ionic detergents (except Tween-40) improved VM-alpha Gal-MC activity by up to 44% but negatively affected VM-alpha Gal-M activity. Both VM-alpha Gal-MC (15% residual activity after 21 min at 85 degrees C, E(d)92.42 kcal/mol) and VM-alpha Gal-M (69.0% residual activity after 10 min at 75 degrees C, E(d)39.87 kcal/mol) showed remarkable thermal stability and repeatedly hydrolyzed the substrate for 10 cycles.