Optimization of Environmental and Nutritional Conditions for the Production of Alkaline Protease by a newly Isolated Bacillus cereus from decaying organic soil

The grain positive bacterium Bacillus cereus has been studied for the production of alkaline protease, an industrially important enzyme. Fermentation was carried out in a shake flask with varying carbon sources (glucose, fructose, sucrose, lactose, mannitol) and nitrogen sources (urea, lysine, ammonium chloride, sodium nitrate, L-glutamic acid) in the broth. The protein and protease enzyme activity was determined for each source after incubation. The production broth was optimized by taking source with highest protein and protease enzyme activity and effect of pH (7, 7.5, 8, 9, 10), temperature (17 degrees C, 27 degrees C, 37 degrees C, 47 degrees C, 57 degrees C) were also optimized. After fermentation crude product was purified by ammonium salt precipitation, osmosis (dialysis) process and ion-exchange chromatography. The molecular mass was determined by gel electrophoresis. The alkaline protease was compatible with four local powdered detergent brands retaining more than 95% activity. Finally, the produced enzyme was immobilized.