Purification and characterization of endo-β-iv-acetyl-glucosaminidase from a flavobacterium sp.

Kenji Yamamoto, Setsu Kadowaki, Kaoru Takegawa1, Hidehiko Kumagai, Tatsurokuro Tochikura

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39 Citations (Scopus)

Abstract

A gram negative bacterium isolated from soil was found to produce a high level of endo-fi-N-acetylglucosaminidase in the culture medium. The organism was identified as a Flavobacterium sp. from various bacteriological characteristics. The enzyme from the Flavobacterium sp. was purified to homogeneity from culture broth by fractionation with ammonium sulfate and column chromatographies on DEAE-cellulose, hydroxylapatite, and Sephadex G-150 and G-100. The molecular weight of the enzyme was estimated to be 27, 000 and 30, 000 by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, and it appeared to consist of a single polypeptide chain. The optimal pH for activity was 5.0 to 6.0 and the stable pH range was 5-7. The Michaelis constant was 0.30 mM with dansyl-Asn-(GlcNAc)2(Man)6 as the substrate. The enzyme hydrolyzed oligosaccharides of native ovalbumin, bovine pancreatic ribonucléase B and a yeast invertase.

Original languageEnglish
Pages (from-to)421-429
Number of pages9
JournalAgricultural and Biological Chemistry
Volume50
Issue number2
DOIs
Publication statusPublished - Jan 1 1986
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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