Purification and characterization of antifungal chitinase from Bacillus safensis MBCU6 and its application for production of chito-oligosaccharides

The chitinase producing bacterial strain was isolated from the vermicompost amended site of Mehsana district of Gujarat, India, and identified as Bacillus safensis MBCU6 using 16S rDNA sequencing. The chitinase was purified by ammonium sulfate precipitation followed by diethylaminoethanol sepharose CL-6B column chromatography. The purified enzyme could be demonstrated as a single band on sodium dodecyl sulfate polyacryalamide gel electrophoresis analysis as well as clear zone on zymogram, with estimated molecular mass of 58 kDa. The optimum pH and temperature of chitinase were pH 7.0 and 60°C, respectively. The purified chitinase exhibited high degree of antifungal activity particularly against pathogenic Macrophomina phaseolina (60%) and Rhizoctonia solani (73%) by dissolving their cell wall components. The purified enzyme could hydrolyze colloidal chitin to its oligomers. It infers that the chitinase produced by Bacillus safensis may play a significant role in the activity as a biopesticide and bioactive material production.