Immobilization of Penicillin G Acylase in Epoxy-Activated Magnetic Cellulose Microspheres for Improvement of Biocatalytic Stability and Activities

Abstract

We prepared magnetic cellulose porous microspheres (MCM) with mean diameter of ∼200 μm by employing the sol−gel transition (SGT) method from a mixture of magnemite ferrofluid and cellulose dissolved in 7 wt % NaOH/12% urea aqueous solvent precooled to −12 °C. Subsequently, the cellulose microspheres were activated with epoxy chloropropane to enhance loading efficiency of biomacromolecules. Their morphology, structure, and properties were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction, and vibrating-sample magnetometer. The results indicated that the spherical magnetic γ-Fe₂O₃ nanoparticles with mean size of 10 nm were uniformly dispersed and embedded in the cellulose substrate of MCM, and the structure and nature of γ-Fe₂O₃ were conserved perfectly. Penicillin G acylase (PGA) as a biocatalyst was immobilized successfully in the porous microspheres, as a result of the existence of the cavity and affinity forces in the activated cellulose matrix. The immobilized PGA exhibited highly effective catalytic activity, thermal stability, and enhanced tolerance to pH variations. Furthermore, the cellulose microspheres loaded with the enzymes could be removed and recovered easily by introducing a magnetic field, leading to an acceptable reusability. Therefore, we have provided a simple and biocompatible support for the enzyme immobilization, which will be promising for the applications in the biomaterial fields.