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http://dx.doi.org/10.4014/mbl.1607.07006

Purification and Characterization of a Chitinolytic Enzyme Produced by Bacillus licheniformis GA9  

Hwang, Dong Ho (Division of Biological Science and Technology, Yonsei University)
Hong, Sung Wook (World Institute of Kimchi)
Hwang, Hyung seo (School of Integrated Oriental Medical Bioscience, Semyung University)
Chung, Kun Sub (Division of Biological Science and Technology, Yonsei University)
Publication Information
Microbiology and Biotechnology Letters / v.44, no.4, 2016 , pp. 470-478 More about this Journal
Abstract
A bacterium producing a large amount of chitinolytic enzyme was isolated from the intestinal tract of earthworm. The isolate was identified as Bacillus licheniformis by 16S ribosomal RNA analysis and designated as B. licheniformis GA9. The enzyme was purified by 40-60% ammonium sulfate precipitation, diethyl-aminoethyl groups exchange chromatography, and gel filtration chromatography. The molecular weight was estimated to be 52.1 kDa and the N-terminal amino acid sequence was D-S-G-K-N-G-K-I-I-R-Y-YP-I-R. The optimum activity of the purified chitinolytic enzyme was shown at pH 5.0 and $40^{\circ}C$, and the enzyme was stable in the ranges of $20-50^{\circ}C$ and pH 5.0-6.0. Enzyme activity was increased by $Co^{2+}$, while it was inhibited by $Cu^{2+}$ and $Fe^{2+}$. But it was recovered by chelating metals with ethylenediaminetetraacetic acid. The $K_m$ and $V_{max}$ values of the purified enzyme were 4.02 mg/ml and 0.52 mg/min, respectively. The chitinolytic enzyme characterized in this study has potential applications in areas such as biotechnology, biomedicine, agriculture, and nutrition.
Keywords
Purification; chitinolytic enzyme; Bacillus licheniformis; GA9;
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