Evaluation of Cotton Seeds as Environmentally Liable Source for Neutral Protease

Proteases are considered one of the most imperative groups of enzymes and are used in bioremediation processes, leather, detergents, pharmaceutical and the food industry. The foremost aim of this study was to extract, purify, and characterize the protease enzyme from cotton seeds. Purification of the neutral protease was achieved with ammonium sulphate, which gave the best results at 60% concentration with a specific activity of 51 units/mg protein and 1.52 purification fold with a percentage yield of 10.5%. The protease was active and stable at a wide range of pH from 4.0–10.0 with an optimum pH of 7.0. The highest activity of the purified enzyme was found at 20 °C. The enzyme was thermally stable and retained 25% of its activity at 50o C. The activity of cottonseeds protease Fraction-IV was enhanced by 20% with ZnCl2 and 15% with CoCl2 as enzyme samples were heated for 10 minutes. The Casein and peptone assays were also performed to check its catalytic activity. Furthermore, the maximum hydrolysis rate (Vmax) and apparent Michaelis–Menten constant (Km) values of the purified protease were 19 μmol/min and 0.08 mol/L, respectively, while activation energy was found to be 12.47 KJ/mol.