Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of the $Exo-{\beta}-D-Glucosaminidase$ from Aspergillus flavus IAM2044

  • Ji, Jae-Hoon (Devision of Appiied Life Science, Graduate School, Gyeongsang National University) ;
  • Yang, Ju-Seok (Devision of Appiied Life Science, Graduate School, Gyeongsang National University) ;
  • Hur, Jong-Wha (Institute of Agriculture and Science, Gyeongsang National University)
  • Published : 2003.04.01
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Abstract

Chitosan-degrading activity induced by chitosan was founf in culture filtrate of Aspergillus flavus IAM2044. Aspergillus flavus IAM2044 had a higher level of chitosanolytic activity when chitosan was used as a carbon source, and yeast extract and peptone were supplemented as nitrogen sources. One of the chitosan-degrading enzymes was purified to homogeneity by ammonium sulfate precipitation followed by cation-exchange and gel filtration chromatographies. The enzyme was monomeric, and its molecular mass was 45 kDa. The optimum pH and temperature of the enzyme were 5.0 and $50^{\circ}C$, respectively. The activity was stable in the pH range of 3.5 to 7.0 and at a temperature below $50^{\circ}C$. Reaction products analyzed by the viscosimetric assay and thin layer chromatography clearly indicated that the enzyme was an exe-type chitosanase, $exo-{\beta}-D-glucosaminidase$, that released GlcN from the nonreducing ends of the oligosaccharide chains.

Keywords

References

  1. FEMS Microbiol. Lett. v.95 Purification and properties of two endochitosanases from Mucor rouxii implicated in its cell wall degradation Alfonso, C.;M. J. Martinez;F. Reyes https://doi.org/10.1111/j.1574-6968.1992.tb05364.x
  2. Proceedings of the First International Conference on Chitin and Chitosan Chitin, an important natural polymer Allan, G. G.;J. R. Fox;N. Kong;R. A. A. Muzzarelli(ed.);E. R. Parsier(ed.)
  3. Annu. Rev. Microbiol. v.22 Cell wall chemisty, morphogenesis, and taxonomy of fungi Bartnicki-Garcia;S. https://doi.org/10.1146/annurev.mi.22.100168.000511
  4. Appl. Microbiol. Biotechnol. v.38 Purification and characterization of a chitosanase from Streptomyces N174 Boucher, I.;A. Dupuy;P. Vidal;W. A. Neugebauer;R. Brzezinski
  5. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the pronciple of protein-dye binding Bradford, M.M. https://doi.org/10.1016/0003-2697(76)90527-3
  6. Eur. J. Biochem. v.1 A protein sequenator Edman, P.;G. Begg https://doi.org/10.1111/j.1432-1033.1967.tb00047.x
  7. J. Gen. Microbiol/ v.126 Purification and mode of action of a chitosanase from Penicillium islandicum Fenton, D. M.;D. E. Eveleigh
  8. Adv. Microb. Ecol. v.11 The ecology of chitin degradation Gooday, G. W. https://doi.org/10.1007/978-1-4684-7612-5_10
  9. Agric. Biol. Chem. v.35 A simple activity measurement of lysozyme Imoto, T.;K. Yagishita https://doi.org/10.1271/bbb1961.35.1154
  10. J. Microbiol. Biotechnol. v.12 Antitumor activity of chitosan oligosaccharides produced in ultrafiltration membrane reactor system Jeon, Y.T.;S.K. Kim
  11. J. Microbiol. Biotechnol. v.11 Effect of antimicrobial activity by chitosan oligosaccharide N-conjugated with asparagines Jeon, Y.T.;S.K. Kim
  12. J. Microbiol. Biotechnol. v.11 Detection of Aspergillus and Penicillium genera by enzyme-linked immunosorbent assay using a monoblonal antibody Kwak, B. Y.;D. H. Shon;B. J. Kwon;C. H. Kweon;K. H. Lee
  13. Nature(London) v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U. K. https://doi.org/10.1038/227680a0
  14. J. Biol. Chem. v.265 Purification and characterization of an exo-β-D-glucosaminidase, a novel type of enzyme, from Nocardia orientalis Nanjo, F.;R. Katsumi;K. Sakai
  15. Appl. Environ. Microbiol. v.64 Purification and characterization of exo-β-D-glucosaminidase from a cellulolytic fungus, Trichoderma reesei PC-3-7 Nogawa, M.;H. Takahashi;A. Kashiwagi;K. Ohshima;H. Okada;Y. Morikawa
  16. Methods Enzymol v.161 Viscometric assay for chitinase Ohtakara, A. https://doi.org/10.1016/0076-6879(88)61051-2
  17. Appl. Environ. Microbiol. v.56 Purification and characterization of three chitosanase activities from Bacillus megaterium P1 Pelletier, A.;J. Syguseh
  18. Biochim. Biophys. Acta. v.1079-65-72 Purification and hydrolytic action of chitosanase, from Nocardia orientalis Sakai, K.;R. Katsumi;A. Isobe;F. Nanjo
  19. Methods Enzymol. v.161 Preparation of crustacean chitin Shimahara, K.;Y. Takiguchi https://doi.org/10.1016/0076-6879(88)61049-4
  20. Appl. Microbiol. Biotechnol. v.54 Molecular cloning and caaracterization of a chitosannase from the chitosanolytic bacterium Burkolderia gladiolo strain CHB101 Shimosaka, M.;Y. Fuikumori;X. Y. Zhang;N. J. He;R. Kodaira;M. Okazaki https://doi.org/10.1007/s002530000388
  21. Biosci. Biotechnol. Biochem. v.57 Chitosanase from the plant parhogenic fungus Fusarium solani f. sp. Phaseoli-purification and some properties Shimosaka, M.;M. Nogawa;Y. Ohno;M. Okazaki https://doi.org/10.1271/bbb.57.231
  22. Appl. Environ. Microbiol. v.61 Production of two chitosanases from a chitosan-assimilatin bacterium, Acinetobavter sp. strain CHB101 Shimosaka, M.;M. Nogawa;X. Y. Wang;M. Kumeara;M. Okazaki
  23. Anal. Biochem. v.178 Detection of chitinase activity after polyacrylamide gel electroporesis Trudel, J.;A. Asslin https://doi.org/10.1016/0003-2697(89)90653-2
  24. Biosci. Biotechnol. Biochem. v.64 Purification and characterization of chitosanase and exo-β-D-glucosaminidase from a koji mold, Aspergillus oryzae IAM2660 Zhang, X. Y.;A.L. Dai;X. K. Zhang;K. Kuroiwa;R. Kodaira;M. Shimosaka;M. Okazaki https://doi.org/10.1271/bbb.64.1896
  25. Chitin, Chitosan. and Related Enzymes Zikakis, J.P.(ed.)