Keratinase are proteolytic enzymes that have gained much attention to convert keratinous wastes that cause huge environmental pollution issues. A new proteolytic Bacillus sp. was isolated from tannery effluent waste soil sample. The bacterium produces extracellular keratinase using horn meal as a sole carbon and nitrogen source. The effect of pH, temperature, and substrate concentration on extracellular keratinase production of this proteolytic strain was studied. Maximum enzyme productions were obtained 681U/mL for 72 hrs of cultivation. The optimal conditions for the proteolytic activity were determined to be pH 8.0 and temperature 40 Â°C; however the proteolytic activity showed a broad range of pH between 6-10 and temperature 30 Â°C â€“ 50 Â°C. The zymogram showed hydrolysis activities of gelatin around the 28 kD. These results indicate that this Bacillus sp. shows a high potential for keratinase production.