The aim of this study is to prepare immobilized protease with high hydrolytic activity forbiotechnological applications. Local Bean Seeds (Dolichos Lablab L.) were used for the extraction of acidprotease using tris-buffer, pH 4.5 as enzyme solvent. Free acidic protease was immobilized on entrapment in calcium alginate gel (in situ activated) by covalent binding method. Their activity and immobilization efficiency for hemoglobin hydrolysis was investigated. Temperature and pH maxima of the immobilized protease showed no changes before and after immobilization. The immobilized protease exhibited good thermal stability and re-usability.