Bio-waste derived dialdehyde cellulose ethers as supports for α-chymotrypsin immobilization

Enzyme immobilization is an important technique to enhance stability, storability and reusability of enzymes. In the present work, pine needles, a forest bio-waste, were used as a feedstock of cellulose to synthesize new materials as supports for immobilization of α-chymotrypsin (CT) enzyme. The extracted cellulose from pine needles was etherified with different alkyl bromides (RBr) and etherified products were further modified to dialdehyde via oxidation with NaIO4 to get the desired products, dialdehyde cellulose ethers (ROcellCHO). CT was then covalently immobilized onto as-synthesized dialdehyde cellulose ethers via Schiff-base formation, i.e., imine linkage. The synthesized products and enzyme immobilization were confirmed by different characterization techniques and the activity assay of the free and the immobilized CT was carried out using standard protocol with variation of different parameters such as temperature, pH and substrate concentration. The storage stability and reusability of the immobilized CT were also investigated. CT activity was also studied in simulated physiological conditions in the artificial gastric fluid and artificial intestinal fluid. Artificial neural network (ANN) model was employed to correlate the relationship with% relative activity and time, temperature and pH affecting enzyme activity. A good correlation of experimental data was predicted by ANN model.