PJB-2020-283
PRODUCTION AND PURIFICATION OF AN EXTRACELLULAR PULLULANASE ENZYME FROM BACILLUS THURINGIENSIS
ASMA ZAFAR
Abstract
In the present study production and purification of an extracellular pullulanase enzyme from Bacillus thuringiensis was carried out for use in various industrial processes. To get the maximum production of pullulanase enzyme, culture conditions like, pH of medium, incubation temperature, time of incubation, media composition and size of inoculum for B. thuringiensis were optimized. Maximum production of pullulanase enzyme (5.71 U/ml/min) was obtained in the medium containing tryptone as carbon and energy source having pH 6.0 when it was inoculated with 3% overnight grown inoculum and incubated at 37°C for 24 hours. Optimal conditions for pullalanse enzyme activity were also determined and maximum enzyme activity (8.584 U/ml/min) was found with 4% pullulan as a substrate in phosphate buffer of pH 7.0 at 50°C after 20 minutes of incubation. Purification of pullulanase enzyme was achieved to homogeneity by ammonium sulphate precipitation as well as by ion exchange chromatography and a distinct band of 97 kDa was analysed by SDS-PAGE. Purification fold of purified enzyme was calculated as 16.83 with 37% yield and 45.45 U/mg specific activity. These results are useful for large scale production of pullulanase enzyme from Bacillus thuringiensis for use in industrial applications.
To Cite this article:
Download
