Abstract

KINETICS AND THERMODYNAMIC STUDIES OF PARTIALLY PURIFIED ALPHA AMYLASE PRODUCED FROM BACILLUS ALTITUDINIS

Pradeep Kumar Singh, Ajay Kumar Singh and Yashab Kumar

ABSTRACT

The present investigation deals with partial purification and kinetics and thermodynamic studies of α- amylase from a strain of Bacillus altitudinis. Amylase is an extracellular enzyme that catalyzes the breakdown of starch into sugars and plays a pivotal role in a variety of areas like ethanol production, detergents, desiring of textiles, modified starches and paper recycling. In the present work, submerged fermentation for α–amylase production has been used. Bacillus altitdunis has been used for the production of α-amylase and was tested using submerged fermentation. Enzyme was extracted by centrifugation at 8,000 rpm for 20 minute. Crude enzyme assays was done by DNS method. Crude enzyme was partially purified by Ammonium sulphate precipitation. After overnight incubation precipitated enzyme was centrifuged at 8000 rpm for 20 minutes at 4oC. After precipitating with ammonium sulphate the partially purified enzyme showed specific activity 0.14 Umg-1 with a total activity of 3.9 Uml-1. In dialysis total enzyme activity was found 1.75Uml-1 specific activity was 0.19 Umg-1.The pH and thermal stability of cellulose free amylases are important issues for their potential industrial applications. In the present research work, the Vmax (8.36 mg/ml/min) and Km (5.46 mg/ml) of α-amylases were derived from the Line-weaver Burke plot. Thermodynamic parameters for soluble starch hydrolysis Ea, ΔH ΔS and ΔG of α-amylase were found to be 37.32KJ/mole, 30.09 KJ/mole,-115.9 J/mole/K and 30442.7 J/mole.

Keywords: ?-Amylase, submerged fermentation, partial purification, Bacillus altitudinis.