Immobilization Horseradish Peroxidase onto UiO-66-NH2 for Biodegradation of Organic Dyes

Enzymes are extensively used as catalyst in several fields of production such as chemistry, and pharmaceuticals owing to their selectivity, efficiency and environmentally friendliness. However, their applications are often hindered due to their insufficient stability and difficulties in re-use. As a member of porous crystalline materials, metal organic frameworks are a promising enzyme carrier due to their multi-functional pore surfaces and robustness in variety of harsh conditions. In this study, the horseradish peroxidase (HRP) enzyme was immobilized onto UiO-66-NH2 (Universitetet i Oslo) by a facile incubation method at the room temperature to improve the stability and reusability of enzyme. The prepared HRP@UiO-66-NH2 bio-composite was characterized by using FT-IR, XRD and SEM. The crystal structure of MOF was well-preserved after enzyme immobilization. A colorimetric assay for enzyme activity after released from UiO-66-NH2 has been employed based on the catalytic oxidation of phenol coupled with 4-aminoantipyrine. The robustness and activity of immobilized enzyme after released from UiO-66-NH2 were investigated by biodegradation of methyl orange (MO) and methylene blue (MB) with several parameters such as pH, temperature, the dosage of H2O2 and the dye concentration with comparison to its free form. The optimum condition for dye degradation was obtained at basic conditions. The immobilized enzyme maintained its activity at elevated temperature while free enzyme lost its activity at the same conditions, attributed to the armoring effect of UiO-66-NH2. According to the results of studied various parameters, MO and MB were biodegraded to 60% and 45%, respectively, within 60 min with the optimum conditions at pH 9 and 50 °C at a H2O2 dosage of 3%. The superior pH tolerance and stability suggest potential of UiO-66-NH2 immobilized peroxidase enzyme in industrial applications.