Isolation, Screening of Alkaline Protease Producing Bacteria and Physio Chemical Characterization of Alkaline Protease

Proteases are a group of enzymes, whose catalytic function is to hydrolyze peptide bonds of proteins and break them down into polypeptides or free amino acids. Large number of bacteria, fungi, Protozoa, Yeast shows proteolytic activities. Soil and waste water sample collected from milk federation effluent area were screened for the presence of alkalophilic bacteria. Isolates were found having proteolytic activity on skimmed milk agar plates. Isolates forming larger zones, as a result of casein hydrolysis were further studied for production of extracellular alkaline protease activity in the shake flask studies. Time course studies indicated that isolate B-5 had the highest protease activity (70.87 µg/ml/min) after 144 h of incubation. The enzyme exhibits maximal activity at 370C, and pH 9 of isolate B-5. The enzyme was purified by ammonium sulphate, dialysis and ion-exchange chromatography using DEAE cellulose column. The isolate B-5 was identified on the basis of morphological, cultural and biochemical Characteristics and was found to be Bacillus subtilis