Skip to main content

Characterization of β-N-Acetylhexosaminidase from rhizostomous jellyfish, Rhopilema asamushi, Mesogloea

Abstract

β-N-Acetylhexosaminidase (EC 3.2.1.52) was purified from rhizostomous jellyfish mesogloea and characterized. Using two purification steps, this enzyme was purified up to 27.4-fold with a recovery rate of 46% compared with crude extract. The molecular weight of the enzyme was estimated to be about 136 kDa, composed of subunit molecular weights of 68 kDa. The enzyme activity was inhibited by SH-reagents, indicating that it contains a SH-group in its active site. The enzyme has a high affinity for pNPGlcNAc with Km value of 0.021 mM. The rate of hydrolysis of N-acetylchito-oligosaccharides tended to decrease with increasing degree of polymerization of the substrate. The parameters of k cat were 92.0 s−1 for pNPGlcNAc, 38.2 s−1 for GlcNAc2, 14.0 s−1 for GlcNAc3, 4.1 s−1 for GlcNAc4, 1.6 s−1 for GlcNAc5, 0.9 s−1 for GlcNAc6, respectively. These results suggest that this β-N-acetylhexosaminidase is an exo-fashion hydrolytic enzyme involved in chitin degradation.

This is a preview of subscription content, access via your institution.

References

  • Bahl, O.P. and Agrawal, K.M.L. 1969. Glycosidases of Aspergillus niger. I. Purification and characterization of α-and β-galactosidase and β-N-acetylglucosaminidase. J. Biol. Chem. 244: 2970–2978.

    PubMed  CAS  Google Scholar 

  • Eriquez, L.A. and Pisano, M.A. 1979. Purification and characterization of an extracellular β-N-acetylhexosaminidase from Paecilomyces persicinus. J. Bacteriol. 137: 620–626.

    PubMed  CAS  Google Scholar 

  • Koga, D., Hoshika, H., Matsushita, M., Tanaka, A., Ide, A. and Kono, M. 1996. Purification and characterization of β-N-acetylhexosaminidase from the liver of a prawn, Penaeus japonicus. Biosci. Biotech. Biochem. 60: 194–199.

    CAS  Google Scholar 

  • Koga, D., Nakashima M., Matsukura, T., Kimura, S. and Ide, A. 1986. Purification and properties of β-N-acetyl-Dhexosaminidase from alimentary canal of the silkworm, Bombyx mori. Agric. Biol. Chem. 50: 2357–2368.

    CAS  Google Scholar 

  • Koga, K., Iwamoto, Y., Sakamoto, H., Hatano, K., Sano, M. and Kato, I. 1991. Purification and characterization of β-N-acetylhexosaminidase from Trichoderma harzianum. Agric. Biol. Chem. 55: 2817–2823.

    PubMed  CAS  Google Scholar 

  • Kramer, K.J. and Koga, D. 1986. Insect chitin. Physical state, synthesis degradation and metabolic regulation. Insect Biochem. 16: 851–877.

    Article  CAS  Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  PubMed  CAS  Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265–275.

    PubMed  CAS  Google Scholar 

  • Nagai, T., Hamada, M., Kai, N., Tanoue, Y., Ueno, S. and Nagayama, F. 1997a. Existence of chitinase and β-Nacetylglucosaminidase in several jellyfish. Fish. Sci. 63: 157–158.

    CAS  Google Scholar 

  • Nagai, T., Hamada, M., Kai, N., Tanoue, Y., Ueno, S. and Nagayama, F. 1997b.Comparison of characteristics of â-Nacetylhexosaminidase from moon jellyfish Aurelia aurita with those from other species. Fish. Sci. 63: 449–452.

    CAS  Google Scholar 

  • Nakagawa, H., Enomoto, N. and Asakawa, M. 1987. Purification and enzymatic properties of β-N-acetylhexosaminidase from common octopus liver. Nippon Suisan Gakkaishi 53: 1025–1031.

    CAS  Google Scholar 

  • Nanjo, F., Ishikawa, M., Katsumi, R. and Sakai, K. 1990. Purification, properties, and transglycosylation reaction of β-Nacetylhexosaminidase from Nocardia orientalis. Agric. Biol. Chem. 54: 899–906.

    CAS  Google Scholar 

  • Reissig, J.L., Strominger, J.L. and Leloir, L.F. 1955. A modified colorimetric method for the estimation of N-acetylamino sugars. J. Biol. Chem. 217: 959–966.

    PubMed  CAS  Google Scholar 

  • Roberts, R.L. and Cabib, E. 1982. Serratia marcescens chitinase: one-step purification and use for the determination of chitin. Anal. Biochem. 127: 402–412.

    Article  PubMed  CAS  Google Scholar 

  • Sakai, T., Nakanishi, Y. and Kato, I. 1993. Purification and characterization of â-N-acetyl-D-hexosaminidase from the midgut gland of scallop (Patinopecten yessoensis). Biosci. Biotech. Biochem. 57: 965–968.

    CAS  Google Scholar 

  • Schlumbaum, A., Mauch, F., Uögeli, U. and Boller, T. 1986. Plant chitinases are potent inhibitors of fungal growth. Nature 324: 365–367.

    Article  CAS  Google Scholar 

  • Sekiguchi, J., Matsumiya, M. and Mochizuki, A. 1995. Distribution of chitinolytic enzymes in seaweeds. Fish. Sci. 61: 876–881.

    CAS  Google Scholar 

  • Ueda, M. and Arai, M. 1992. Purification and some properties of β-N-acetylglucosaminidase from Aeromonas sp. 10S-24. Biosci. Biotech. Biochem. 56: 1204–1207.

    Article  CAS  Google Scholar 

  • Yamamoto, K., Lee, K.M., Kumagai, H. and Tochikura, T. 1985. Purification and characterization of â-N-acetylhexosaminidase from Penicillium oxalicum. Agric. Biol. Chem. 49: 611–619.

    CAS  Google Scholar 

Download references

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nagai, T., Watarai, Y. & Suzuki, N. Characterization of β-N-Acetylhexosaminidase from rhizostomous jellyfish, Rhopilema asamushi, Mesogloea. Fish Physiology and Biochemistry 25, 53–59 (2001). https://doi.org/10.1023/A:1019707816787

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1019707816787

  • characteristics
  • comparison
  • jellyfish
  • life cycle
  • purification
  • β GlcNAcase