Journal of Microbiology and Biotechnology

  • 제9권6호
  • /
  • Pages.839-846
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  • 1999
  • /
  • 1017-7825(pISSN)
  • /
  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Purification and Characterization of a Chitinase from Cytophaga sp. HJ Isolated from Sea Sand

  • Lee, Dong-Mi (The Research Institute of Pharmaceutical Sciences and College of Pharmacy, Ewha Womans University) ;
  • Noh, Hee-Jung (The Research Institute of Pharmaceutical Sciences and College of Pharmacy, Ewha Womans University) ;
  • Lee, Kang-Man (The Research Institute of Pharmaceutical Sciences and College of Pharmacy, Ewha Womans University)
  • 발행 : 1999.12.01
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초록

An extracellular chitinase-producing bacterial strain induced by colloidal chitin was isolated from sea sand and was identified to be a member of the genus Cytophaga. The chitinase was purified successively by 30-60% ammonium sulfate fractionation, and DEAE-Bio gel A column, Octyl-Sepharose CL-4B column, and DEAE-Bio gel A column chromatographies. The enzyme had a molecular mass of 59.75 kDa, and the amino terminal amino acid sequence was ATPNAPVISW MPTDXXLQNXS. The enzyme acted better on colloidal chitin as a substrate than on chitosan. For colloidal chitin and chitosan (Degree of Acetylation, 15-25%), $K_{cat}$ values were 0.60U/mg and 0.08U/mg, respectively. HPLC analysis of the enzymatic reaction products showed that the chitinase produced mostly N-acetyl-D-glucosarnine and di-N-acetylchitobiose. The optimum temperature and pH for the enzyme were $50^{\circ}C$ and 4.0, respectively. N-Bromosuccinimide and $Hg^{2+}$ inhibited the chitinase activity as much as 90%, and $Sb^{3+}$, diethylpyrocarbonate, and $Ag^{+}$ inhibited it by 50-70%.

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