Isolation and Purfication of Xylanase from Bacillus Cereus by Submerged State Fermentation

Sivasankari P¹*, Flory Shobana M^2
1Department of Biotechnology, PGP College of Arts & Science, Namakkal, Tamil Nadu, India
²Department of Biotechnology, Sree Narayana Guru College, Coimbatore, Tamil Nadu, India

ABSTRACT
Enzymes are important classes of globular proteins of biological origin that act as biochemical catalysts. Xylanase catalyses the hydrolysis of complex sugars (xylan) to simpler sugars (xylose) and also important in bioconversion of hemicellulose into the constituents sugar. Xylanases shows great potential for industrial applications mainly for the bioconversion of lignocelluloses to sugars, ethanol and other substances, clarification of juices and wines, improving the nutritional quality of silage and green feed and the deinking process of waste papers. The soil samples were collected and the Bacillus cereus was isolated by spread plate method. The isolated organism was screened by xylan agar medium. The fermentation media was characterized based on effect of substrate feeding, effect of temperature, effect of initial pH, effect of incubation time and effect of inoculums. At optimised medium the pure culture was inoculated and shake flask and bioreactor were carried out. The germinated crude extract was collected and the enzyme activity was estimated. The crude enzyme was purified by ammonium sulphate precipitation, dialysis and ion exchange chromatography and the molecular mass of enzyme was determined by SDS PAGE. The purified enzyme was immobilized by sodium alginate entrapment method.
Keywords: Bacillus cereus, Xylanases, SDS PAGE