KINETICS AND THERMODYNAMICS APPROACH FOR IMMOBILIZATION OF ALPHA-AMYLASE AND MALTOGENIC AMYLASE BY NANOMAGNETIC COMBINED CROSS-LINKED ENZYME AGGREGATES METHOD FOR STARCH CONVERSION TO MALTOSE SYRUP

The thermostability enhancement of specified nanomagnetic combi-CLEAs of α-amylase and maltogenic amylase was evaluated by assessment of kinetic parameters, temperature dependence, enzyme half-life, thermal inactivation energy barriers (Ea(in)), and thermodynamic parameters in a storage thermostability approach at 55–95° C for 120 minutes. The Michaelis constant (Km) values of NM-Combi-CLEAs was diminished within 1.5 folds while The catalytic efficiency was enhanced near 1.32 after CLEAs formation. The half-lives of the fabricated nano-bio catalyst increased about 2.5 folds at 95º C. Besides, Ea(in) of the native combined enzymes was appended from 44.75 into 54.76 (KJ.mol-1) and ΔH* increased from 41.69 to 51.70 (KJ.mol-1) while, ΔS* decreased 28.2% after immobilization. The NM-Combi-CLEAs conserved 80.4% of its original activity after ten cycles. Hence, the advanced technique for NM-Combi-CLEAs production is introduced as an effective and secured method to fabrication of recyclable immobilized enzyme also, resolving of mass transfer limitation difficulties and ease in enzyme handling.