Proteases are the most useful industrial enzymes. The protease has versatile applications including extensive uses in confectionary, fermentation process, leathering, detergent industry,etc. In this research Enterococcushirae, a gram positive bacteriawas enriched and immobilized in various matrices to study their protease producing efficiency. The immobilization was carried out by three different matrices under two different concentrations of 3% and 4%. Repeated batch fermentation using the immobilized cells were carried out in boiling tube with Skim Milk as the substrate and their protease producing efficiency were studied. The results showed that the 4% calcium alginate had the maximum enzyme activity of  283.2 (U/ml) and also maintained a good level of enzyme production throughout the entire 10 days cycle. The 4% calcium alginate also showed consistent protease production with the enzyme activity consistently maintained above 150 (U/ml). The average protease production by the microorganism is 176.64 (U/ml). The peak activities for other methods areas follows: 3% calcium alginate – 249.6 (U/ml), 3% agar-agar – 235.2 (U/ml), 4% agar-agar – 230.4 (U/ml), 3% Gelatin – 235.2 (U/ml) and 4% Gelatin – 174.4 (U/ml). The entrapment of cells in 4% calcium alginate was found to be the optimal method for immobilizing Enterococcus hirae.