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Purification and Characterization of Chitinase from Paenibacillus illinoisensis KJA-424  

JUNG WOO JIN (Glucosamine Saccharide Materials-National Research Laboratory (GSM-NRL), Division of Applied Bioscience and Biotechnology Institute of Agricultural Science and Technology, Chonnam National University)
KUK JU HEE (Glucosamine Saccharide Materials-National Research Laboratory (GSM-NRL), Division of Applied Bioscience and Biotechnology Institute of Agricultural Science and Technology, Chonnam National University)
KIM KIL YONG (Glucosamine Saccharide Materials-National Research Laboratory (GSM-NRL), Division of Applied Bioscience and Biotechnology Institute of Agricultural Science and Technology, Chonnam National University)
KIM TAE HWAN (Department of Animal Science, College of Agriculture and Life Science, Chonnam National University)
PARK RO DONG (Glucosamine Saccharide Materials-National Research Laboratory (GSM-NRL), Division of Applied Bioscience and Biotechnology Institute of Agricultural Science and Technology, Chonnam National University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.2, 2005 , pp. 274-280 More about this Journal
Abstract
A chitinase was purified from the culture supernatant of Paenibacillus illinoisensis KJA-424 by protein precipitation, DEAE-Sephadex anion-exchange chromatography, and Sephadex G-150 gel filtration. The molecular weight of the purified chitinase was 54 kDa on SDS-PAGE and activity staining. Optimal pH and temperature were pH 5.0 and 60$^{circ}$C, the presence of 10 ruM Ag$^{+}$ and Hg$^{2+}$ inhibited the activity by $92.1/%$ and $97.7/%$, and the K$_{m}$ and V$_{max}$ values were 1.12 mg chitin mrl and 1.48$\mu$mol GlcNAc min$^{-1}$, respectively. The enzyme hydrolyzed tetramer to dimer, pentamer to dimer and trimer, and hexamer to dimer, trimer and tetramer, indicating an endo-splitting mechanism. The chitinase had no hydrolytic activity toward dimer and trimer. The chitinase inhibited the mycelial growth of Rhizoctonia solani, suggesting an antifungal property.
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