Evaluation of Coupling Protocols to Bind Beta-Glucosidase on Magnetic Nanoparticles

Abstract

Beta-Glucosidase has been chosen as a model biomolecule to establish a general protocol for binding enzymes on both ferromagnetic and superparamagnetic nano-particles for sensing applications. Using EDC (1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide) or SMCC (Succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate) as coupling agents, we compared two different methods for the fabrication of enzyme-decorated magnetic nanoparticles. We identified the best conditions for the preparation of a responsive bioactive magnetic system comparing different covalent bio-grafting protocols. The enzymatic test has been performed using beta-Glucosidase. The systems were characterized using scanning electron microscopy, infrared spectroscopy, and the enzyme loading was measured by a glucose assay in the presence of the enzyme-decorated magnetic particles. Although the faster response of ferromagnetic particles to the magnetic field, the assay results suggested that the superparamagnetic particles are more efficient carriers. In fact, the best enzymatic activity was measured on superparamagnetic systems that have the further advantage of preventing aggregation induced by the residual magnetization. Hence, beta-Glucosidase coated magnetic nanospheres could provide an attractive system suitable for the cleavage and the rapid evaluation of glycoside levels in natural products, measuring the liberated glucose without the need for specialised instrumentation. Moreover, the magnetic particles allow the subsequent collection of enzymes for further analysis, such as its use in portable fast screening kits or devices.

Hybrid Nanomaterials of Magnetic Iron Nanoparticles and Graphene Oxide as Matrices for the Immobilization of β-Glucosidase: Synthesis, Characterization, and Biocatalytic Properties

“…More specifically, the highest immobilization efficiency was observed for bgl on iron-based nanomaterials, rather than on functionalized GO. Similar immobilization efficiencies have been previously reported for bgl on magnetic nanoparticles (Ricco et al, 2014) or GO-iron nanoparticles hybrid nanomaterials (Gokhale et al, 2013;Ricco et al, 2014). It is interesting to note that the type of amine groups on the surface of the hybrid nanomaterial seems to affect the immobilization efficiency.…”

<p>Magnetically Directed Enzyme/Prodrug Prostate Cancer Therapy Based on β-Glucosidase/Amygdalin</p>

“…β-Glu has been successfully immobilized on magnetic nanoparticles and showed favourable activity. 37,38 In a previous study, we also prepared β-Glu-loaded magnetic nanoparticles and confirmed their targeted delivery using a mouse model of subcutaneous 9L glioma. 39 Moreover, polyethylene glycol (PEG) modification of β-Glu increased the in vivo stability of MNP-β-Glu and enhanced their accumulation in the targeted tumor tissue.…”