In this study, hydrazine treated acrylic fabrics (polyacrylonitrile, PAN) activated with cyanuric chloride was developed as supporting material for horseradish peroxidase (HRP) immobilization. The immobilization of HRP onto the modified supporting material was achieved after being end-over-end incubated for 12 h. Field emission scanning electron microscopy and Fourier-transform infrared spectroscopy techniques were used to confirm the successful immobilization. Reusability experiment was performed to estimate the ability of the immobilized HRP to recover the reaction medium, in which it was observed to retain 78% of its original activity after 10 cycles. Relative to the soluble HRP, the optimum pH and temperature for the immobilized HRP were shifted to 7-7.5 and 50 °C, respectively. The kinetic parameters of guaiacol and H2O2 for the immobilized HRP were determined to be Km/Vmax = 57.61, 11.35 and Kcat/Km = 1.87, 1.86, respectively, while the values for the free form were Km/Vmax = 41.49, 6.23 and Kcat/Km = 1.87, 1.86, respectively. Compared to the soluble form, the immobilized HRP exhibited higher resistance toward metal ions and some organic solvents. For an application perspective. The immobilization of HRP using this procedure has the potential to be used for industrial application and wastewater treatment.
Keywords: Horseradish peroxidase; Hydrazine hydrochloride; Immobilization.
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