New approach to the immobilization of glucose oxidase on nonporous silica microspheres functionalized by (3-aminopropyl)trimethoxysilane (APTMS)

Abstract

The immobilization and encapsulation of glucose oxidase (GOD) onto the mesoporous and the nonporous silica spheres prepared by co-condensation of tetraethylorthosilicate (TEOS) and (3-aminopropyl)trimethoxysilane (APTMS) in the W/O emulsion system were studied. The terminal amine group was used as the important functionality for GOD immobilization on the silica substrate. When only TEOS is used as a silica source, the disordered mesoporous silica microspheres are obtained as our previous results. As the mole ratio of APTMS to TEOS (RAT) increases, the surface area and pore volume of the silica particles measured by nitrogen adsorption and desorption method and SEM decrease rapidly. Particularly, the largest change of the surface morphology is observed between RAT = 0.20 and RAT = 0.25. The amount and the adsorption time of immobilized enzyme were measured by UV spectroscopy. About 20 wt % of GOD was immobilized into the silica substrates above RAT = 0.60 and was completely adsorbed into the substrate of RAT = 0.80 4h after addition. In the measurement of the thermal stability, GOD dissolved in buffer solution loses nearly all of its activity after 30 min at 65°C. In contrast, GOD immobilized on the surface-modified silica particles still retains about 90% of its activity after the same treatment. At this temperature, the immobilized glucose oxidase retained half of its initial activity after 4h. (Figure 1) It is shown that the suitable usage of fuctionalizing agent like APTMS as well as the control of surface morphology is very important on the immobilization of enzyme.