EDTA-Cu (II) chelating magnetic nanoparticles as a support for laccase immobilization

Developments in nanotechnology have led to the discovery of new materials, namely, magnetic nanoparticles (MNPs), that present easy surface functionalization and high surface-to-volume ratios. These properties allow a high mass transfer rate and easy removal from a reaction matrix. Simple separation under an external magnetic field makes them a promising immobilization support for enzymes. In this work, new MNPs were prepared by functionalization with EDTA-TMS and characterized by TEM, FTIR and BET analytical techniques, among others. These MNPs were applied as support for laccase immobilization to create a promising biocatalyst. Despite the known chelating nature of EDTA-TMS, its use for surface modification of MNPs for laccase immobilization is a rather unexplored strategy and is reported here for the first time. At pH 3.5, the immobilization process showed approximately 97% of enzymatic activity recovery. The Michaelis-Menten kinetic properties of immobilized laccase showed a lower Vmax and a similar KM compared to free laccase. Regarding operational stability, the immobilized enzyme presented approximately 73% of its initial activity after five sequential reactive cycles. The immobilized enzyme was successfully applied to the biocatalysis of Indigo Carmine dye degradation. These MNPs with immobilized laccase showed important advantages compared to other materials for application in industrial biochemical processes, biocatalysis and biosensors.