Production, Optimization, Purification and Characterization of Mannanase Produced by Bacillus Subtilis

Abstract

Endo-β-1,4-D-mannanase (β-mannanase; EC 3.2.1.78) catalyses the random hydrolysis of mannoglycosidic bonds in mannan-based polysaccharides. These enyzmes are commonly found in nature and are located within the structure of mannans and heteromannans (galactomannan, glucomannan and galactoglucomannan) in the hemicellulose fraction of trees with soft tissues and hard tissues, locust bean seed.  Most β-mannanases degrade mannooligosaccharides down to a degree of polymerization of four. In this study mannanase was produced using a submerged fermentation method from Bacillus subtilis. The effect of growth of the organism and processing parameters on the production of mannanase were determined after which optimization studies were carried out. The enzyme was partially purified using ammonium sulphate, dialysis and gel filtration. The partially purified enzyme was characterized. The result of the study showed that mannanase enzyme from Bacillus subtilis was optimally produced in a medium comprising of galactose, peptone, sugarcane bagasse at a pH of 6.0 and a temperature of 45 oC. The enzyme was most stable and active at pH 10.0 and at a temperature of 40 oC.  and 60 respectively.