RESUMO
Bacillus megaterium 314 strain was able to utilize agricultural and industrial wastes for metallo-protease production. Orange peel and wheat bran were found as the most suitable carbon and nitrogen sources, respectively. Optimized production process enhanced the enzyme production by 5.1-folds. Glass and glass-ceramic with different particle sizes based on mica were used as inorganic carrier. Protease enzyme was immobilized by covalent bonding and physical adsorption methods on nanoparticle supports. Enzyme physically adsorbed on glass ceramic (particle size 0.71-1.0 mm) had the highest residual activity and the highest immobilization yield. Glass-ceramic was characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). Immobilized enzyme exhibited activation energy (E a ) and deactivation rate constant at 60 °C (k d ) about 1.29 and 1.46-times, respectively lower than free enzyme. Moreover, adsorbed enzyme had higher energy for denaturation (E d ), half-life (t 1/2 ), and decimal reduction time (D). The thermodynamic parameters of irreversible thermal denaturation for the protease enzyme indicate that immobilized enzyme had higher enthalpy (ΔH°), free energy (ΔG°), and entropy (ΔS°) than free one. There was a significant improvement in the maximum reaction velocity Vmax (2.5-fold), Michaelis constant Km (1.9-fold), and catalytic efficiency Vmax/Km (4.7-fold) values after immobilization indicating the efficiency and effectiveness of immobilization approach.
