Journal of Pure and Applied MicrobiologyVol. 7 No. 1

Production, Purification and Characterization of b-glucosidase from Bacillus subtilis strain PS Isolated from Sugarcane Bagasse

A.K. Verma*, Soniya Saini, S. Nishad, Vinod Kumar, Shalini Singh and Ashutosh Dubey

Department of Biochemistry, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar- 263 145, India.

Received on 21 July 2012 and accepted on 11 September 2012

 

ABSTRACT

The effect of various carbon sources on extracellular b-glucosidase (BGL) production and its catalytic activity with different substrate was analyzed in Bacillus subtilis strain PS isolated from sugarcane bagasse. The highest extracellular enzyme activity was obtained in minimal medium with supplement of glucose was higher 0.42 U/ml, compared to other carbon sources such as maltose, cellobiose, lactose and xylose. Extracellular BGL was partially purified to 18.04 fold with specific activity 17.32 U/mg using Sephadex G-75 gel filtration chromatography after dialysis of 70% ammonium sulfate precipitation. In gel filteration fractions, three peaks having b-glucosidase activity and represents isozymes of b-glucosidase with molecular weight of 193 kDa, 64 kDa and 42 kDa, respectively. Purified BGL showed catalytic activity with lactose, cellobiose, pNPG and mellibiose.

Keywords : b-Glucosidase production; Gel filteration; Carbon source; Bacillus subtilis.