Partial purification and characterization of chitinase produced by Bacillus licheniformis B307

Abstract

The optimal conditions required for chitinase production from Bacillus licheniformis B307 strain, obtained from Syrian soil, were studied. Optimization experiments were carried out under submerged fermentation conditions, and colloidal chitin was the source of carbon. Luria broth medium supplied with 0.5% colloidal chitin was the optimum medium for chitinase production. The maximum chitinase yield was obtained at 30 °C, pH6, incubation time 14 days, and 150 rpm. The optimum chitinase activity was achieved at 60 °C and pH6. The chitinase activity with unmodified medium was 1.9 U/mL which then enhanced about eight folds to reach 14.2 U/mL under optimized submerged fermentation conditions. An extracellular chitinase of Bacillus licheniformis B307 was partially purified using ammonium sulfate precipitation followed by concentration with various sizes of concentrator tubes. The chitinase was partially purified 8.24 fold and specific enzyme activity increased 2.08 fold (2 U/mg). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of partial purified chitinase exhibited a molecular weight ( M _r) near to 36 and 42kDa. These results make it possible to invest in this strain to produce chitinase to be used as antifungal, food additives and other applications.

Abstract

Biochemistry; Biotechnology; Microbiology; Molecular biology; Plant biology; Chitinase; Bacillus licheniformis; Optimization; Submerged culture; Purification