Biopolymers conjugated with magnetite as support materials for trypsin immobilization and protein digestion

Abstract

In the presented study synthesized magnetic nanoparticles were used as an inorganic precursor for the preparation of novel magnetite-lignin and magnetite-chitin hybrid supports for enzyme immobilization. Effective synthesis of the hybrids was confirmed by Fourier transform infrared spectroscopy and powder X-ray diffraction analysis. The materials exhibited good thermal stability and surface areas of 4.3 and 5.6 m²/g respectively. The magnetite-lignin + trypsin and magnetite-chitin + trypsin systems were found to have good storage stability and reusability. After 20 days they retained over 75% and 90% respectively of their initial activity, and after 10 consecutive biocatalytic cycles retained over 60% and 80% respectively of their initial activity. The kinetic parameters of the free and immobilized enzyme were also comprehensively examined and compared. The results of peptide digestion tests confirmed the high proteolytic activity of the produced trypsin-based magnetic biocatalytic systems.

Keywords: Enzyme immobilization; Hybrid supports; Magnetic materials; Peptide digestion; Trypsin.