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Covalent immobilization of chloroperoxidase onto magnetic beads: Catalytic properties and stability

Paper ID Volume ID Publish Year Pages File Format Full-Text
4565 232 2008 9 PDF Available
Title
Covalent immobilization of chloroperoxidase onto magnetic beads: Catalytic properties and stability
Abstract

Amino groups containing magnetic beads were used in covalent immobilization of the enzyme “chloroperoxidase (CPO)” which is one of a few enzymes that can catalyse the peroxide dependent oxidation of a wide spectrum of organic and inorganic compounds. The magnetic poly(glycidylmethacrylate-methylmethacrylate-ethyleneglycol dimethacrylate), magnetic p(GMA-MMA-EGDMA) beads were prepared via suspension polymerization in the presence of ferric ions. The magnetic beads were characterized with scanning electron microscope (SEM), Fourier transform infrared (FTIR), Mössbauer spectroscopy and vibrating sample magnetometer (VSM). The magnetic beads were derivatized sequentially with ammonia and glutaraldehyde, and CPO was covalently immobilized on the support via reaction of the amino groups of the enzyme under mild conditions. The effect of various parameters including pH, temperature and enzyme concentration on the immobilization efficiency of CPO onto glutaric dialdhyde activated magnetic beads was evaluated. Magnetic measurement revealed that the resultant CPO-immobilized magnetic beads were superparamagnetic with a saturation magnetization of 18.2 emu/g. The analysis of FTIR spectra confirmed the binding of CPO on the magnetic beads. The maximum amount of immobilized CPO on the magnetic beads was 2.94 mg/g support. The values of Michaelis constants Km for immobilized CPO was significantly larger, indicating decreased affinity by the enzyme for its substrate, whereas Vmax values were smaller for the immobilized CPO. However, the CPO immobilized on the magnetic beads resulted in an increase in enzyme stability with time.

Keywords
Magnetic beads; Enzyme immobilization; Chloroperoxidase; Kinetic parameters; Enzyme stability
First Page Preview
Covalent immobilization of chloroperoxidase onto magnetic beads: Catalytic properties and stability
Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 38, Issue 2, 15 February 2008, Pages 180–188
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering