Production, purification and enzyme kinetics of amylase from bacteria isolated from a soil of Solapur, Maharashtra state, India

Author(s): Sunita Bansod*, Deepak Thigale, Priyanka Dure, Harshada Sohani

Abstract

Amylase is the enzyme which is used for various purposes. So our aim was to isolate the amylase producing Bacillus subtilis spp. from soil. In present work we isolated various Bacillus spp. from soil and studied the physical parameters like pH, temperature, and substrate concentration of amylase enzyme. We got four Bacillus isolates from soil, which produce amylase. One isolate was selected which shows larger zone of starch hydrolysis. Amylase production was studied by using iodine solution and partial characterization of bacillus was carried out by studying the morphological characterization. As amylase is extra cellular enzyme it can easily extracted by centrifugation and purification was carried out by salt precipitation and dialysis technique. The purified amylase enzyme activity was determined and also the different parameters of enzyme such as pH, temperature and substrate concentration were studied, the activity of amylase enzyme isolated from “A” is 480 μl/ml of enzyme/min. It’s shown maximum activity at pH 6.5 and shown maximum activity at 40°C. Also conclude that the rate of reaction increases with increasing substrate concentration up to 0.19 in which the rate of reaction remains constant. The point of saturation is obtained at 0.19 optical density, hence the value of Vmax and Km value from graph is Vmax = 0.19 O.D, Km = 9.5 O.D (concentration- 0.7mg).

Share this article

International Journal of Bioassays is a member of the Publishers International Linking Association, Inc. (PILA), CROSSREF and CROSSMARK (USA). Digital Object Identifier (DOI) will be assigned to all its published content.