Purification and Characterization of Bacterial Chitinase isolated from Crustacean Shells

Four Chitin degrading bacterial strains vibrio aestuarianus, Flavobacterium, Shewenella and Exiguobacterium were isolated from crustacean shells. The strains were confirmed by Biochemical analysis, FAME-GC analysis and 16s rDNA sequencing. The chitinase was purified by a two step chromatographic method and characterized. The enzyme was purified to homogeneity by 6.75 fold with 46% recovery after ion exchange chromatography followed by gel filtration chromatography. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular mass of 24 kDa. It showed an optimum pH at 6.0. The optimum temperature for enzyme activity was 40 °C. The maximum activity was observed with a 2% substrate concentration of colloidal chitin. The enzyme was strongly inhibited by Fe2+ and K+ while enhanced by Zn2+ and Ca2+. Thus the purification of microbial chitinase from shell waste could be effectively utilized for the manufacturing of many chitin derived products.