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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.
Keywords: Chitinase, Crustacean shells, FAME-GC, ion exchange chromatography, gel filtration
chromatography, Characterisation.