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A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
Published on: November 7, 2012
Characterization and application of a detergent-stable alkaline α-amylase from Bacillus subtilis strain AS-S01a
Jetendra K Roy1, Sudhir K Rai, Ashis K Mukherjee
1Department of Molecular Biology and Biotechnology, School of Science and Technology, Tezpur University, Tezpur, Assam, India.
Abstract:
A strain AS-S01a, capable of producing high-titer alkaline α-amylase, was isolated from a soil sample of Assam, India and was taxonomically identified as Bacillus subtilis strain AS-S01a. Optimized α-amylase yield by response surface method (RSM) was obtained as 799.0 U with a specific activity of 201.0 U/mg in a process control bioreactor. A 21.0 kDa alkaline α-amylase purified from this strain showed optimum activity at 55°C and pH 9.0, and it produced high molecular weight oligosaccharides including small amount of glucose from starch as the end product. The K(m) and V(max) values for this enzyme towards starch were determined as 1.9 mg/ml and 198.21 μmol/min/mg, respectively. The purified α-amylase retained its activity in presence of oxidant, surfactants, EDTA and various commercial laundry detergents, thus advocating its suitability for various industrial applications.
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