Epoxy-Affixed ZIF-8/CS/Cellulase: a Sustainable Approach for Hydrolysis of Agricultural Waste to Reducing Sugars.

Cellulase was effectively immobilized onto an epoxy-bound chitosan-modified zinc metal-organic framework (epoxy/ZIF-8/CS/cellulase) support, yielding a conjugation rate of 0.64 ± 0.02 mg/cm2 and retaining 80.01 ± 0.01% of its specific activity. The bare and cellulase-bound supports was characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, atomic force microscopy and energy-dispersive X-ray spectroscopy. The immobilized enzyme exhibited optimal activity at pH 5.5 and a temperature of 70 ℃. The efficiency, stability and reactivity of the enzyme improved after immobilization, as evidenced by a decrease in activation energy, enthalpy and Gibbs free energy along with an increase in entropy change. The epoxy-affixed ZIF-8/CS/cellulase strip was successfully employed for rice husk hydrolysis achieving an impressive conversion efficiency of 95%. The method demonstrated a linear range from 0.1 to 0.9% (0.1 × 10 to 0.9 × 10 mg/ml) and exhibited a strong correlation (R = 0.998) with the widely adopted 3, 5-dinitrosalicylic acid method. The epoxy/ZIF-8/CS bound cellulase exhibited remarkable thermal stability, retaining 100% of its activity at 70 °C, in contrast to just 53% for the free enzyme and displayed a half-life of 21 days after storage at 4 °C compared to 9 days for the free enzyme. Furthermore, it retained over 95% activity after 12 h at pH levels of 4.5 and 5.5 and showcased excellent reusability, maintaining activity over 25 cycles. Overall, this method offers high conversion efficiency and selectivity under benign conditions, with no undesirable by-products, making it a cost-effective solution for the routine hydrolysis of lignocellulosic biomass feedstock.