Abstract: Following the enrichment technique in total 25 microbial colonies were isolated, purified, preserved and tested for their proteolytic ability. Among the bacterial isolate D1 showed the maximum ability to degrade egg albumin, milk casein and gelatin. On the basis of cultural, morphological, physiological and biochemical characteristics the isolate D1 was identified as Bacillus fastidiosus den Dooren de Jong. The isolate (D1) produced maximum protease at 37 ° C temperature and pH 7.0. Cellobiose (carbon source) and (NH4)2HPO4 (nitrogen source) were found to induce the protease production of the isolate and showed highest proteolytic activity at temperature 35 ° C and pH 6.5.