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A study on partial purification and characterization of extracellular amylases from Bacillus subtilis

Jahir Alam Khan and Ruchika Priya

Soil collected from kitchen waste decomposing plots of the restaurant, Lucknow was enrinched in laboratory by adding chopped potatoes to enhance the Amylase producing microorganisms. The bacterial isolate isolated from the enriched soil was screened for amylase production, and the isolate showing maximum hydrolysis identified as Bacillus subtilis. The production was carried out by submerged fermentation. Maximum Amylase activity (0.0115 U/ml/min) was obtained at 37°C after 72 h of incubation. The enzyme was purified using ammonium sulphate precipitation and showed a molecular weight of 65Kda by SDS- PAGE. The enzyme was relatively stable between pH 7 to pH 9 and at temperature ranging from 37°C to 50°C. Moreover, activity was enhanced by using metal ions such as Ca2+, Mg2+ and Zn2+. The decline in activity was observed by adding SDS and EDTA.