Innovative immobilization of β-glucosidase on amidoximated acrylic fabric integrated with magnetite Fe₃O₄...

This research presents a novel system for immobilizing β-glucosidase on amidoximated acrylic fabric integrated with magnetite nanoparticles. Amidoximated acrylic fabric (TAC) was prepared using hydroxylamine hydrochloride, and magnetite nanoparticles were incorporated to create TAC-Fe₃O₄, which was subsequently activated with glutaraldehyde and used to immobilize β-glucosidase. The immobilization efficiency reached an 89 % yield. Characterization techniques such as FTIR, SEM, and BET analysis confirmed successful immobilization, whereas kinetic studies revealed an increase in Km (7.83 mM for TAC- Fe₃O₄@β-Glu vs. 5.45 mM for free enzyme). The free β-glucosidase exhibited maximum activity at pH 5.5, while TAC-Fe₃O₄@β-Glu demonstrated an optimal pH of 6.0. Similarly, the free enzyme showed maximum activity at 40 °C, whereas the immobilized enzyme achieved its maximum activity at 50 °C. Operational stability tests revealed that TAC-Fe₃O₄@β-glucosidase retained 79 % of its initial activity after 10 reuse cycles and 72 % after 15 cycles. Storage stability studies demonstrated a significant advantage for the immobilized enzyme, which maintained 85 % of its activity after 8 weeks at 4 °C, compared with 52 % for the free enzyme. These results highlight the potential of TAC-Fe₃O₄ as a cost-effective and scalable system for enzyme immobilization, offering improved stability and reusability, making it highly suitable for industrial applications.