Immobilization of horseradish peroxidase on amidoximated acrylic polymer activated by cyanuric chloride.
Horseradish peroxidase (HRP) was immobilized on amidoximated acrylic fabric after being activated with cyanuric chloride. FT-IR spectroscopy and scanning electron microscopy were used to characterize fabrics. The maximum immobilization efficiency of HRP (70%) was detected at 4% cyanuric chloride and pH 7.0. The immobilized enzyme retained 45% of its initial activity after ten reuses. The immobilization of enzyme on the carrier is saturated after 6h of incubation time. The pH was shifted from 7.0 for soluble HRP to 7.5-8.0 for the immobilized enzyme. The soluble HRP and immobilized HRP had the same optimum activity at 40°C. The immobilized HRP is more thermal stable than soluble HRP. Substrate analogues were oxidized by immobilized HRP with higher efficiencies than those of soluble HRP. Km values of the soluble HRP and the immobilized HRP were 31 and 37mM for guiacol and 5.0 and 7.8mM for H2O2, respectively. The immobilized HRP had higher efficiency for removal of phenol than that of soluble HRP. The immobilized HRP had higher resistance toward heavy metal ions compared to the soluble enzyme. The immobilized HRP was more stable against high concentration of urea, Triton X-100 and isopropanol. The immobilized HRP exhibited high resistance to proteolysis by trypsin than soluble enzyme. In conclusion, the immobilized HRP could be used as a potential efficient catalyst for the removal of aromatic pollutants from wastewater.
Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia; Molecular Biology Department, National Research Center, Dokki, Cairo, Egypt. Electronic address: saleh38@hotmail.com.
Saleh A
SA
Mohamed
Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
Saeed S
SS
Al-Ghamdi
Chemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia; Dyeing, Printing and Textile Auxiliaries Department, Textile Research Division, National Research Center, Dokki, Cairo, Egypt.
Reda M
RM
El-Shishtawy
D000180:Acrylic Resins / Q000737:chemistry*; D000577:Amides / Q000737:chemistry*; D004795:Enzyme Stability / Q000187:drug effects; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008670:Metals / Q000494:pharmacology; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature; D013997:Time Factors; D014227:Triazines / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism
D000180:Acrylic Resins; D000577:Amides; D004800:Enzymes, Immobilized; D007477:Ions; D008670:Metals; D014227:Triazines; C006903:carbopol 940; C019309:cyanuric chloride; D006735:Horseradish Peroxidase; D014357:Trypsin
Acrylic polymer; Immobilization; Peroxidase