Preparation and characterization of hydrogels for selective separation of β-glucosidase enzyme

In the first step of the thesis study, β-glucosidase imprinted hydrogels were prepared using monomers having different functions such as acrylamide (AA), hydroxyethylmethacrylate (HEMA), 1-vinylimidazole (1-VI), 4-vinylpyridine (4-VP), methacrylic acid. The imprinting efficiencies of the hydrogels were compared and it was seen that the hydrogel with the highest imprinted efficiency was prepared with the acrylamide monomer. Structural characterization of hydrogel prepared for the selective separation of β-glucosidase enzyme was performed by Fourier Transform Infrared Spectroscopy (FTIR) analysis and surface morphology analysis by scanning electron microscopy (SEM). Effect of incubation time, pH temperature and enzyme concentration were examined in order to optimize the rebinding conditions of β-glucosidase enzyme on MIP and NIP hydrogels. As a result of experimental studies, optimum rebinding time of enzyme on MIP and NIP hydrogels was determined as 120 min, optimum rebinding temperature as 25 ° C and optimum pH 7.0. When the initial enzyme concentration reached 2.5 mg / mL, the adsorption reached saturation. From these data, adsorption isotherms like Langmuir and Freundlich isotherms were formed. It is seen that the Freundlich isotherm model describes the MIP and NIP hydrogel adsorption better than the Langmuir isotherm model. β-Glucosidase adsorption capacities of MIP and NIP hydrogels were calculated as 7.2 mg / g and 4.6 mg / g, respectively. Kinetic analysis of the experimental data showed that the pseudo second order model represented the best adsorption kinetics. The decrease in adsorption with temperature showed that the adsorption was exothermic. The swelling properties of cross-linked hydrogels were studied related time. It was observed that the rate of swelling of both hydrogels reached equilibrium at the end of 90 minutes and the maximum swelling ratio was calculated as 24.79% and 16.43%. The selectivity of β-glucosidase imprinted hydrogel was measured by cross-recognition tests with different enzymes (α-Amylase and lipase). The availability of β-glucosidase imprinted hydrogel prepared as the affinity material was tested at the end of the thesis.