Purification and Production Optimization of Alpha Amylases from Bacillus Species

Abstract

Amylases are the digestive enzymes belong to the family of glycoside hydrolase capable of hydrolysing starch and polysaccharides. These are the recent breakthroughs in food, feed, pulp and pharma industries. Our aim of this study is to purify amylases from Bacillus subtilis and Bacillus megaterium and optimize the growth conditions to achieve the cost-effective production of amylases from bacillus species. Extensive research on amylases from different sources is essential in producing enzymes in large scale and in optimizing the growth conditions and minimize the production cost to meet the increased demand of these industrially important enzymes. Amylases were produced in the current study using submerged fermentation and purified the extracellular enzymes from bacillus species using ultracentrifugation, Ammonium sulphate precipitation and gel filtration chromatography. There was no effect on amylase production from bacillus species when the initial substrate concentration is increased from 1% to 5%. Increase in inoculum size from 1% to 5% and the addition of yeast extract to broth had significantly increased the amylase production. The study observed significantly higher levels of amylase production using rice starch when compared with wheat starch also higher levels of amylase production observed with B. megaterium when compared with B. subtilis. The study concludes 1% rice starch and 5% B. megaterium inoculum can be used for cost effective production of amylases. In the current study, cost effective amylase production was evaluated and able to optimize the yield of extracellular amylases from Bacillus megaterium to a concentration level of 120 IU/ml of fermentation broth.