Immobilization of horseradish peroxidase on activated wool

• Published in: Process biochemistry (1991) Vol. 48; no. 4; pp. 649 - 655
• Main Authors: Mohamed, Saleh A., Darwish, Anas A., El-Shishtawy, Reda M.
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01.04.2013
• Subjects: Activated; butanol; Chlorides; Cyanuric chloride; denaturation; Enzymes; Fourier transform infrared spectroscopy; Horseradish; hydrogen peroxide; Immobilization; immobilized enzymes; isopropyl alcohol; metal tolerance; octoxynol; Optimization; Peroxidase; proteolysis; scanning electron microscopy; temperature; trypsin; urea; Wool; Horseradish; Immobilization; Cyanuric chloride; Peroxidase; Wool
• ISSN: 1359-5113; 1873-3298
• DOI: 10.1016/j.procbio.2013.03.002

•A new activated wool has been prepared using cyanuric chloride.•Horseradish peroxidase (HRP) was successfully immobilized on activated wool.•The results revealed that the immobilized HRP has enhanced physical and chemical properties.•The wool-HRP was more stable to several denaturants. This work is aimed to immobilize partially purified horseradish peroxidase (HRP) on wool activated by multifunctional reactive center, namely cyanuric chloride. The effect of cyanuric chloride concentration, pH and enzyme concentration on immobilization of HRP was studied. FT-IR and SEM analyses were detected for wool, activated wool and immobilized wool-HRP. The wool-HRP, prepared at 2% (w/v) cyanuric chloride and pH 5.0, retained 50% of initial activity after seven reuses. The wool-HRP showed broad optimum pH at 7.0 and 8.0, which was higher than that of the soluble HRP (pH 6.0). The soluble HRP had an optimum temperature of 30°C, which was shifted to 40°C for immobilized enzyme. The soluble and wool-HRP were stable up to 30 and 40°C after incubation for 1h, respectively. The apparent kinetic constant values (Kms) of wool-HRP were 10mM for guiacol and 2.5mM for H2O2, which were higher than that of soluble HRP. The wool-HRP was remarkably more stable against proteolysis mediated by trypsin. The wool-HRP exhibited more resistance to heavy metal induced inhibition. The wool-HRP was more stable to the denaturation induced by urea, Triton X-100, isopropanol, butanol and dioxan. The wool-HRP was found to be the most stable under storage. In conclusion, the wool-HRP could be more suitable for several industrial and environmental purposes.