Isolation and identification of Bacillus producing thermophilic alpha amylase: production optimization and investigation of the activity and stability of enzyme.

Abstract

Alpha-amylases are the most important amylases used in industry. Among them, thermophilic alpha-amylases are of particular importance, which is due to their activity and stability in high temperatures. These enzymes produced by thermophile micro-organisms including bacteria. These thermophilic alpha-amylases are used in various industries such as processing of starch as well as production of detergents and biofuels. In this research, the bacteria which produce the thermophilic alpha-amylases were isolated and characterized in hot springs of Gorooh village in Kerman province. According to the results of screening on the specific liquid and solid media, AT59 was selected as the best strain. Morphological and biochemical characterization of the isolated strain indicated that it belonged to Bacillus sp. and was gram-positive, catalase positive, casein hydrolyzing and acid producing from lactose and sucrose. The results obtained from the optimization of the enzyme production medium showed that among the carbon, nitrogen and ion sources investigated, starch (1 gr/l), gelatin (2 g/l) and magnesium sulfate (1 g/l) had the most increasing effect on the production of AT59 alpha-amylase. Moreover, the highest enzyme production was obtained at pH 5. This enzyme also demonstrated the highest degree of activity and stability in 80 and 70 ℃, respectively. These findings suggested that this enzyme has a considerable potential for use in starch industry.

Keywords

• hot spring
• production
• screening
• thermophilic enzymes