Abstract

Present study dealt with the production, purification and characterization of xylanase from Bacillus subtilis- BS05 grown in submerged fermentation at 37^oC for 48h using sugarcane bagasse as a substrate. Xylanase enzyme was purified to homogeneity by ammonium sulphate fractionation followed by sephadex G-100 chromatography. The molecular mass of xylanase enzyme was found to be 23kDa by SDS-PAGE. The optimum pH and temperature of xylanase enzyme was 5 and 50^oC with stability range of 5-6 and 30^oC -50^oC respectively. The enzyme had half life of 1386-1155 minutes at 30^oC -50^oC respectively. Metal profile of the enzyme showed that Mn²⁺ (127.4 %) and Fe²⁺ (115%) were the activators while Hg²⁺ (14%) and EDTA (19%) were the inhibitors. Kinetic parameters revealed that the enzyme is specific to birchwood xylan with K_m , K_cat , V_max and K_cat/K_m value of 1.15 mg/ml, 850 s^-1, 117.64 U/mg and 739.13 s^-1mg^-1.mL respectively.

Keywords: Xylanase, purification, characterization, Bacillus sp.