Preparation of p-aminophenol modified superparamagnetic iron oxide nanoparticles for purification of alpha-amylase from the bovine milk

Abstract

Currently, modified Fe3O4 magnetic nanoparticles frequently are used as nanocarriers for proteins and enzymes purification. In the present study, Fe3O4 magnetic nanoparticles were prepared, and their surfaces were modified with p-aminophenol affinity ligand immobilized by different linkers. The modified nanocarriers were used for the purification of alpha-amylase from the bovine milk (after precipitation the casein) by affinity purification. To evaluate the effectiveness of the p-aminophenol modified magnetic nanocarriers, the three different types of nanocarriers with different linkers having varying lengths were prepared. All nanocarriers were characterized and validated using FT-IR, SEM, EDX, VSM and XRD analysis methods According to our results, p-aminophenol ligand attached to the nanocarrier by long linkers better separates the alpha-amylase from the casein free skim milk with 49.66% recovery and 48.18-fold purification efficiency. The results of this study showed that our novel magnetic nanocarriers have the capacity to be used for fast, reproducible and cost-effective purification of alpha-amylase.