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Purification and characterization of a new glucoamylopullulanase from thermotolerant alkaliphilic Bacillus subtilis DR8806 of a hot mineral spring

Paper ID Volume ID Publish Year Pages File Format Full-Text
34575 45034 2012 10 PDF Available
Title
Purification and characterization of a new glucoamylopullulanase from thermotolerant alkaliphilic Bacillus subtilis DR8806 of a hot mineral spring
Abstract

We have introduced a novel glucoamylopullulanase from thermostable alkaliphilic Bacillus subtilis DR8806 from a hot mineral spring in Iran. The enzyme was purified by ion-exchange chromatography following to ammonium sulphate precipitation. The molecular weight of the purified enzyme was estimated to be 65.5 kDa using denaturing acrylamide gel electrophoresis. The enzyme showed high activity over a wide pH range, from pH 5.0 to pH 11.0 with the optimum pH 9.5. Our results also indicated an optimum temperature of the enzyme activity at 70 °C. These features justify the characteristics of the alkaliphilic and thermostable bacterial proteins and enzymes. The enzyme did not require calcium and showed extreme stability with regard to surfactants, including SDS and Triton X-100, and oxidizing agents such as H2O2. These features of the enzyme suggest a promising potential for application in laundry industry. Furthermore, the enzyme was active on pulullan by 68% relative to normal activity on starch. Such characteristics have not already been reported for this type of enzyme, hence we propose that this is a new alkalophilic and thermostable enzyme.

► We purified a new glucoamylase from a thermostable alkaliphilic Bacillus subtilis. ► A 65.5 kDa enzyme with optimum of activity at pH 9.5 and 70 °C was characterized. ► This calcium-independent enzyme showed extreme stability against some surfactants. ► The enzyme is active on pulullan by 68% relative to normal activity on starch.

Keywords
Bacillus subtilis; Hot-spring; Glucoamylopullulanase; Purification; Alkaline-enzyme; Thermotolerant
First Page Preview
Purification and characterization of a new glucoamylopullulanase from thermotolerant alkaliphilic Bacillus subtilis DR8806 of a hot mineral spring
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 47, Issue 5, May 2012, Pages 806–815
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering