Thermal deactivation of α-amylase immobilized magnetic chitosan and its modified forms: A kinetic and thermodynamic study

The stability and thermodynamic properties of an enzyme are the main factors that governing its applications in industry. With that intention we have immobilized the α-amylase onto synthesized chitosan-magnetite (CSM) composite and its modified forms by gluteraldehyde (CSM-GLA), glyoxal (CSM-GLY) and epichlorohydrin (CSM-ECH). In this study all the immobilized enzymes exhibited improved pH stability about 60–80% of relative activity at pH 9 compared to the free enzyme. The temperature stability at 60 °C is up to 50% of relative activity for covalently immobilized enzymes as enzyme become more rigid by covalent binding and so protected from the conformational changes caused by the environment. The thermal deactivation of the free and immobilized enzymes follows the first order kinetics. The t1/2 and D-values were prolonged considerably in case of covalently immobilized enzymes, indicating better thermal stability than that of free and adsorbed ones. The Ed values 18.71, 32.00, 27.19 and 20.46 KJmol-1 for CSM-E, CSM-GLY-E, CSM-GLA-E and CSM-ECH-E described the high stability and resistance to heat inactivation. The Km values 0.525 ± 0.04, 0.57 ± 0.06, 0.65 ± 0.04 mg/mL and Vmax values 25 ± 0.06, 19.6 ± 0.02, 16.39 ± 0.01 μmol mg−1 min−1for CSM-GLY-E, CSM-GLA-E and CSM-ECH-E showing better substrate affinity. The immobilized enzymes have exhibited about 60% of relative activity after 90 days of storage and very good reuse potential.