Polystyrene microspheres with a small amount of acrylic acid (200:1) incorporated were synthesized by nonaqueous phase dispersion (NAD) polymerization to investigate the feasibility of use as a carrier for immobilized enzyme. Seeded emulsion polymerization was then carried out to obtain larger cross-linked polystyrene spheres, the structure of which was expected to yield a favorable environment to keep the original activity of the immobilized enzyme. Glucoamylase, a particular enzyme catalyzing the hydrolysis of oligosaccharides to glucose, was immobilized through peptide bonding using a carbodiimide activating agent. Thirty percent of the enzyme activity at maximum was retained relative to free glucoamylase. The immobilized enzyme remained active after 4 months, although its poor resistance to heat was not improved.