Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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)
  • Published : 1999.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

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%.

Keywords

References

  1. Science v.212 Chitin: New faces of research Austin, P. R.;C. J. Brinel;J. E. Castle;J. P. Zikakis
  2. J. Gen. Microbiol. v.138 Chitinase and chitin synthase 1: Counterbalancing activities in cell separation of Saccharomyces cerevisiae Cabib, E.;S. J. Silverman;J. A. Shaw
  3. Int. J. Biol. Macromol. v.8 New method for the quaternization of chitosan Domard, A.;M. Rinaudo;C. Terrassin
  4. J. Biochem. v.105 Purification and characterization of chitinase produced by Streptomyces erythraeus Hara, S.;Y. Yamamura;Y. Fujii;T. Mega;T. Ikenaka
  5. Bergey's Manual of Determinative Bacteriology(9th ed.) Holt, J. G.;N. R. Krieg;P. H. A. Sneath;J. T. Staley;S. T. Williams
  6. Agr. Biol. Chem. v.35 A simple activity measurement of lysozyme Imoto, T.;K. Yagishita
  7. Meth. Enzymol. v.8 Chitinases Jeuniaux, C.
  8. EMBO J v.5 Isolation and characterization of genes encoding two chitinase enzymes from Serratia marcescens Jones, J. D. G.;K. L. Grady;T. V. Suslow;J. R. Bedbook
  9. Food Technol. v.38 Use of chitinous polymers in food - A challenge for food research and development Knorr, D.
  10. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U. K.
  11. J. Microbiol. Biotechnol. v.7 Purification and characterization of antifungal chitinase from Pseduomonas sp. YHS-A2 Lee, H.-S.;H.-J. Lee;S.-W. Choi;S. Her;D-H. Oh
  12. J. Biol. Chem. v.193 Protein measurement with the folin phenol reagent Lowry, O. H.;N. J. Rosebrough;A. L. Farr;R. J. Randall
  13. Biosci. Biotech. Biochem. v.56 Purification and characterization of a novel type of chitinase from Vibrio alginolyticus TK-11 Murao, S.;T. Kawada;H. Itoh;H. Oyama;T. Shin
  14. Chitin Muzzarelli, R. A. A.
  15. J.Agr. Food Chem. v.37 Crawfish chitosan as a coagulant in recovery of organic compounds from seafood processing streams No, H. K.;S. P. Meyers
  16. J. Bacteriol. v.174 Chitinase of Streptomyces olivaceoviridis and significance of processing for multiplicity Romaguera, A.;U. Menge;R. Breves;H. Diekmann
  17. Biochim. Biophys. Acta v.83 The hydrolysis of chitin by concentrated hydrochloric acid, and the preparation of low-molecular-weight substrates for lysozyme Rupley, J. A.
  18. Appl. Environ. Microbiol. v.60 Purification and characterization of thermostable β-N-acetylhexosaminidase of Bacillus stearothermophilus CH-4 isolated from chitin-containing compost Sakai, K.;M. Narihara;Y. Kasama;M. Wakayama;M. Moriguchi
  19. J. Microbiol. Biotechnol. v.5 Isolation and characterization of Acinetobacter sp. WC-17 producing chitinase Shin, W.-C.;D.-S. Lee;T.-H. Kim;J-H. Woo;J.-M. Lee;J.-G. Kim;S.-D. Hong
  20. Bergey's Manual of Systematic Bacteriology Stanley, J. T.;M. P. Bryant;N. Pfennig;J. G. Holt
  21. Carbohyd. Polym. v.24 Chitinases of Streptomyces kurssanovii: Purication and some properties Stoyachenko, I. A.;V. P. Varlamov;V. A. Davankov
  22. Biochim. Biophys. Acta v.1078 Isolation and characterization of thermostable chitinases from Bacillus licheniformis X-7u Takayanagi, T.;K. Ajisaka;Y. Takiguchi;K. Shimahara
  23. Chitin. Rev. Pure Appl. Chem. v.7 Tracey, M. V.
  24. Appl. Environ. Microbiol. v.59 Purification and properties of a thermostable chitinase from Streptomyces thermoviolaceus OPC-520 Tsujibo, H.;K. Minoura;K. Miyamoto;H. Endo;M. Moriwaki;Y. Inamori
  25. Can. J. Microbiol. v.38 Purification, properties, and partial amino acid sequence of chitinase from a marine Alteromonas sp. strain O-7 Tsujibo, H.;Y. Yoshida;K. Miyamoto;C. Imada;Y. Okami;Y. Inamori
  26. Biosci. Biotechnol. Microbiol. v.59 Purification and some properties of six chitinases from Aeromonas sp. No. 10S-24 Ueda, M.;A. Fujiwara;T. Kawaguchi;M. Arai.
  27. J. gen. Microbiol. v.137 Regulation of chitinase synthesis in Trichoderma harzianum Ulhoa, C. J.;J. F. Peberdy
  28. J. Biol. Chem. v.265 Gene cloning of chitinase A1 from Bacillus circulans WL-12 revealed its evolutionary relationship to Serratia chitinase and to the type III homology units of fibronectin Watanabe, T.;K. Suzuki;W. Oyanagi;K. Ohinshi;H. Tanaka
  29. J. Gen. Appl. Microbiol. v.32 Purication and characterization of chitinase and chitinase and chitobiase produced by Aeromonas hydrophila subsp. anaerogenes A52 Yabuki, M.;K. Mizushina;T. Amatatsu;A. Ando;T. Fujii;M. Shimada;M. Yamashita
  30. J. Microbiol. Biotechnol. v.8 Cloning and expression of a chitinase gene from Thermoactinomyces vugaris KFB-C100 Yoon, H.-G.;H.-Y. Kim;Y.-H. Lim;H.-Y. Cho
  31. J. Microbiol. Biotechnol. v.8 Cloning and expression of a chitinase gene from Thermoactinomyces vulgaris KFB-C100 Yoon,H.G.;H.Y.Kim;Y.H.Lim;H.Y.Cho