Cloning and expression of Recombinant antifungal chitinase enzyme of Bacillus pumilus in Bacillus subtilis 168

  Introduction: Chitin, a linear polysaccharide and is one of the most abundant natural polysaccharides is the main component in numerous of organisms. Chitinase enzymes are essential enzymes catalyzing the conversion of chitin to its monomeric or oligomeric components, therefore, chitinase can used as a natural antifungal agent. The purpose of this study is cloning and expression of recombinant chitinse gene in Bacillus subtilis in the treatment offungal infection effects and replacement with carcinogenic and synthetic products.   Materials and Methods : Chitinase gene of Bacillus.pumilus was cloned to integrate vector Pdh5 Through designed the appropriate pimers.built construct using dual crossing over for transferred into the amylase gene (amyE) of Bacillus subtilis168 and its expression was induced by IPTG and its function was evaluated.   Results : Recombinant construct containing chitinase gene was produced. Gene expression by creating halo around the recombinant bacteria colonies in minimal media including chitin as carbon source was observed.   Conclusion: Create a halo containing chitin in the minimal media represents the performance of recombinant chitinase enzyme to catalyze the chitin. Considering antifungal property can be effectively treated as a matter of human fungal infection and safe substance to excretion biological pest and pollutants.