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Distribution of chitinases and characterization of two chitinolytic enzymes from one-year-old Korean Ginseng (Panax ginseng C.A. Meyer) roots

  • Moon, Jong-Kook (Department of Molecular Science and Technology, Graduate School, Ajou University) ;
  • Han, Beom-Ku (Avicore Biotechnology Institute, Optifarm Solution Inc.) ;
  • Kim, T. Doo-Hun (Department of Molecular Science and Technology, Graduate School, Ajou University) ;
  • Jo, Do-Hyun (Department of Molecular Science and Technology, Graduate School, Ajou University)
  • Received : 2010.07.05
  • Accepted : 2010.09.11
  • Published : 2010.11.30

Abstract

We report the tissue-specific distribution of chitinolytic activity in Korean ginseng root and characterize two 31-kDa chitinolytic enzymes. These two enzymes (SBF1 and SBF2) were purified 70- and 81-fold with yields of 0.75 and 1.25%, respectively, and exhibited optimal pH and temperature ranges of 5.0-5.5 and 40-$50^{\circ}C$. With [$^3H$]-chitin as a substrate, $K_m$ and $V_{max}$ values of SBF1 were 4.6 mM and 220 mmol/mg-protein/h, respectively, while those of SBF2 were 7.14 mM and 287 mmol/mg-protein/h. The purified enzymes showed markedly less activity with p-nitrophenyl-N-acetylglucosaminide and fluorescent 4-methylumbelliferyl glycosides of D-N-acetylglucosamine oligomers than with [$^3H$]-chitin. End-product inhibition of both enzymes demonstrated that both are endochitinases with different N-acetylglucosaminidase activity. Furthermore, the $NH_2$-terminal sequence of SBF1 showed a high degree of homology with other plant chitinases whereas the $NH_2$-terminal amino acid of SBF2 was blocked.

Keywords

References

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