Immobilization of horseradish peroxidase on modified chitosan beads.
A method has been developed to immobilize horseradish peroxidase (HRP) on modified chitosan beads by means of graft copolymerization of polyethylacrylate in presence of potassium persulphate and Mohr's salt redox initiator. The activity of free and immobilized HRP was studied. FTIR spectroscopy and scanning electron microscopy were used to characterize HRP immobilization. The efficiency of the immobilization was investigated by examining the relative enzymatic activity of free enzyme before and after the HRP immobilization. The obtained values were found to reach 98.4%. The results show that the optimum temperature of immobilized HRP was 45 degrees C, which was identical to that of free enzyme, and the immobilized HRP exhibited a higher relative activity than that of free HRP over 45 degrees C. The optimal pH for immobilized HRP was 10, which was higher than that of the free HRP (pH 9.0), and the immobilization resulted in stabilization of enzyme over a broader pH range. The apparent kinetic constant value (K(m)) of immobilized HRP was 3.784 mmol ml(-1), which was higher than that of free HRP. On the other hand, the activity of immobilized HRP decreased slowly against time when compared to that of the free HRP, and could retain 65.8% residual activity after 6 consecutive cycles.
Monier
M
M
Chemistry Department, Drexel University, Philadelphia, PA, USA. monierchem@yahoo.com
Ayad
D M
DM
Wei
Y
Y
Sarhan
A A
AA
D000180:Acrylic Resins / Q000737:chemistry; D055162:Biocatalysis; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties; D013696:Temperature
D000180:Acrylic Resins; D004800:Enzymes, Immobilized; C478961:poly(ethylacrylate); D048271:Chitosan; D006735:Horseradish Peroxidase