Amylase Production and Purification from Bacteria Isolated from a Waste Potato Dumpsite in District Farrukhabad U.P State India

Starch is an abundant carbon source in nature, and α-amylase (1, 4-a-D-glucanohydrolase; EC 3.2.1.1), which hydrolyzes a-1, 4-glucosidic linkage in starch-related molecules, is one of several enzymes involved in starch degradation. Alpha amylase is a hydrolytic enzyme and in recent years, interest in its microbial production has increased dramatically due to its wide spread use in food, textile, baking and detergent industries. The soil samples were obtained (10gm) from Farrukhabad district, U. P. state, India. The samples were analyzed for bacteriological study. One gram of (1.0 gm) soil sample was inoculated in to a liquid soluble starch medium generated reducing sugar with a minimum concentration of 1.35 mg/ml after 48 hours of incubation. The soluble starch amylases were characterized and it was revealed that optimum temperature of activity was 400C. The optimum pH activity was observed between 7.5-8.5. The optimum substrate concentration for bacterial growth was 0.4% w/v with activity of 10.5IU/ml. The most effective precipitation of enzyme took place at 60% w/v salt concentration. Bacillus subtilis was found to be most frequently occurring amylolytic bacteria followed by Bacillus cerus and Bacillus megaterium. The mean zone of amylolytic activity for the isolates ranged between 1.8 mm for Bacillus subtilis and 0.9 mm for Bacillus cereus. The activity of the purified enzyme was 17 IU/ml. The purpose of the current investigation was isolation, production and purification of Bacillus species isolated from soil in order to study their suitability with regard to α-amylase production.