Hydrogel matrices based on starch and dimethylaminoethyl methacrylate (Starch/DMAEMA) were synthesized including γ-irradiation as a clean initiator. The prepared hydrogels were characterized in terms of their gel fraction yield, degree of equilibrium swelling. The prepared hydrogels were examined as carriers for immobilization of Bacillus subtilis that has the ability to secrete an extracellular fibrinolytic enzyme that degrades fibrin. Scanning electron microscope (SEM) analysis showed proliferation of the bacterial cells entrapped inside the polymeric matrix. The immobilization process increases the production time of fibrinolytic enzyme up to 120 h instead of 96 h for the free cells. The optimum temperature of activity broadened and a significant shift in the pH optima was observed upon immobilization. The reusability of immobilized cells under repeated batch fermentation conditions was also investigated. At the optimum production conditions, immobilization of B. subtilis cells onto Starch/DMAEMA resulted in a four fold increase in enzyme activity.