JOURNAL OF APPLIED POLYMER SCIENCE, cilt.101, sa.2, ss.818-824, 2006 (SCI-Expanded)
The uniform and nonporous poly(glycidyl methacrylate) (poly(GMA)) latex particles, 1.7 mu m in size, were first tried as a support in enzyme immobilization. For this purpose, alpha-chymotrypsin (CT) was selected as the model enzyme. The low particle size and nonporous character of the selected support allowed to carry out the enzyme-subtrate interaction on a sufficiently large surface area (3.36 m(2)/g) and in the absence of intraparticular diffusion limitations. This property is particularly important when the immobilized CT is used for the substrates with high molecular weights (i.e., proteins). The latex particles were synthesized by dispersion polymerization of GMA. The reactive character of poly(GMA) allowed the direct attachment of primary amine groups onto the particles. Confocal laser scanning microscopy (CLSM) showed that primary amine groups were preferentially located on the particle's surface. Hence, the selected enzyme, CT was immobilized on the surface of nonporous particles via glutaraldehyde activation. For CT-immobilized poly(GMA) particles, the maximum activity (r(m)) and Michealis constant (K-m) were found to be 17.2 mu mol/mg CT min and 121.6 mu m, respectively. No significant loss was observed in the activity of immobilized CT, during the course of experiments. (c) 2006 Wiley Periodicals, Inc.