Immobilization of horseradish peroxidase on cationic microporous starch: Physico-bio-chemical characterization and removal of phenolic compounds

Abstract

In the present study, two different modified starches; microporous starch (MPS) and cationic microporous starch (CMPS) were synthesized. The granules of MPS that distributed regularly were destroyed after the etherification reaction. The data depicted that the immobilization of horseradish peroxidase (HRP) on CMPS revealed highest immobilization efficiency (86%) at 100 mg of CMPS at pH = 6.0 and 100 units of enzyme. After 10 reuses of the CMPS-HRP, it retained 66% of initial activity. The soluble HRP showed broad pH optimum of 6.0-7.0, which changed to sharp pH = 6.0 for CMPS-HRP. Soluble-HRP and CMPS-HRP showed temperature optima at 30 °C and 40 °C, respectively. The CMPS-HRP showed high thermal stability up to 50 °C compared to the soluble HRP (40 °C). The Km values of soluble HRP and CMPS-HRP were 6.6 and 10.8 mM for H₂O₂ and 34 and 41.6 mM for guaiacol, respectively. CMPS-HRP showed higher affinity toward various substrates than the soluble-HRP. CMPS-HRP showed more resistance against heavy metals, urea, isopropanol, Triton X-100 and trypsin than soluble enzyme. The CMPS-HRP showed higher ability to remove phenol and p-chlorophenol compared to soluble-HRP.

Keywords: Enzyme immobilization; Modified starch; Removal of phenol and p-chlorophenol.