Preapration of mono size nanobeads and its usage for lysozyme adsorption

In this study, poly(2-hydroxyethyl methacrylate) P(HEMA) nanoparticles carrying methacryloylamidophenylalanine (MAT) ligand, as a comonomer, providing hydrophobic functionality to the adsorbent were prepared. MAT was synthesized by reacting methacryloyl chloride with phenylalanine. We selected P(HEMA) as the basic solid matrix. Nanospheres with an average size of 155.64 nm were obtained by emulsion polymerization of P(HEMA) and MAT. The nanoparticles had a specific surface area of 1051.3 m2/g, and were characterized by elemental analysis and Fourier Transform Infrared Spectroscopy (FTIR). Nanospheres which synthesized and characterized were used for the lysozyme immobilization. Adsorption experiments were investigated under different medium conditions (i.e., medium pH, initial concentration, temperature) in a batch system. Lysozyme adsorption capacity of P(HEMA) and P(HEMA-MAT) beads were 1.2 mg/g and 770.0 mg/g respectively. In addition to these, studies of optimization of the immobilization conditions were obtained. Kinetic parameters (Vmax, Km) of the immobilized enzyme were determined and compared to the kinetic parameters that of free enzyme. It was observed that after 5 adsorption-desorption cycle, P(HEMA-MAT) nanoparticles can be used without significant loss in lysozyme adsorption capacity.