Bacterial proteases are considered potential candidates for industrial and biotechnological applications. In the present study, the influence of various parameters on an in-house isolated alkaline protease was investigated through central composite design from a well-known response surface methodology technique. A high level of alkaline protease productivity up to 8.9-folds was achieved from a novel strain of Bacillus subtilis isolated from local Tannery, Lahore Distt. (Pakistan). The purification profile revealed 8.48-fold increase to homogeneity by ammonium sulfate fractionation and Sephadex G-100-based gel filtration chromatography. The purified fraction was identified as a monomeric with apparent molecular weight of 15 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The catalytic characterization was performed with various substrates using Michaelis-Menten equation. The alkaline protease remained optimally active within a broader pH range of 8-12 having a half-life of 37.8 min at 87 A degrees C, along with a maximum velocity (V-max) of 200 IU/mL/min and 0.090 mg/mL Michaelis constants. Moreover, a complete de-staining and dehairing were recorded within the short incubation period. In conclusion, the data thus obtained in this study suggest a high potential for enzymatic treatment in an industrial setting.