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Purification and characterization of chitinase enzyme from Kurthia gibsonii Mb126

Journal of Environment & Biotechnology Research, Vol. 2, No. 1, Pages 37-44, January 2016

Mini K.Paul, K.D. Mini, Jyothis Mathew

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ABSTRACT

The aim of the study was to purify and characterize the chitinase enzyme from Kurthia gibsonii Mb126. The chitinase enzyme from K. gibsonii Mb126, the chitinolytic bacterial strain, was purified through four steps including ammonium sulphate precipitation, affinity adsorption, ion exchange chromatography and gel filtration chromatography. The chitinase was purified 16.11-fold through Sepadex G 100 gel filtration. The specific activity of the purified enzyme was 10.31 U/mg proteins after purification. The purified enzyme on analysis with Coomassie Brilliant Blue R 250 gave a single band near 40 KDa indicating the homogeneity of the preparation. The enzyme was most active at pH 6.5.  The optimum temperature for the chitinase was 40^◦ C. Impacts of various metal ions, chemicals and detergents were studied. The pH stability and thermo stability of the chitinase were also studied.  The chitinase exhibited K_m and V_max values of 11.1mg/mL and 11.12 µmoles/µg h, respectively. To the best of our knowledge, it is the first report on characterization of chitinase from Kurthia.
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