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

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

Research output: Contribution to journalArticle

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

Fingerprint

Flavobacterium
Hexosaminidases
Purification
Enzymes
enzymes
beta-Fructofuranosidase
DEAE-Dextran
Acetylglucosaminidase
DEAE-Cellulose
Column chromatography
ovalbumin
Ovalbumin
Ammonium Sulfate
Durapatite
beta-fructofuranosidase
Fractionation
Electrophoresis
Gram-Negative Bacteria
Oligosaccharides
Gram-negative bacteria

All Science Journal Classification (ASJC) codes

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

Cite this

Purification and characterization of endo-β-iv-acetyl-glucosaminidase from a flavobacterium sp. / Yamamoto, Kenji; Kadowaki, Setsu; Takegawa, Kaoru; Kumagai, Hidehiko; Tochikura, Tatsurokuro.

In: Agricultural and Biological Chemistry, Vol. 50, No. 2, 01.01.1986, p. 421-429.

Research output: Contribution to journalArticle

Yamamoto, Kenji ; Kadowaki, Setsu ; Takegawa, Kaoru ; Kumagai, Hidehiko ; Tochikura, Tatsurokuro. / Purification and characterization of endo-β-iv-acetyl-glucosaminidase from a flavobacterium sp. In: Agricultural and Biological Chemistry. 1986 ; Vol. 50, No. 2. pp. 421-429.
@article{24e841f83d9643a08c14b0e4322d7df2,
title = "Purification and characterization of endo-β-iv-acetyl-glucosaminidase from a flavobacterium sp.",
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 ribonucl{\'e}ase B and a yeast invertase.",
author = "Kenji Yamamoto and Setsu Kadowaki and Kaoru Takegawa and Hidehiko Kumagai and Tatsurokuro Tochikura",
year = "1986",
month = "1",
day = "1",
doi = "10.1080/00021369.1986.10867399",
language = "English",
volume = "50",
pages = "421--429",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "2",

}

TY - JOUR

T1 - Purification and characterization of endo-β-iv-acetyl-glucosaminidase from a flavobacterium sp.

AU - Yamamoto, Kenji

AU - Kadowaki, Setsu

AU - Takegawa, Kaoru

AU - Kumagai, Hidehiko

AU - Tochikura, Tatsurokuro

PY - 1986/1/1

Y1 - 1986/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0009743199&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0009743199&partnerID=8YFLogxK

U2 - 10.1080/00021369.1986.10867399

DO - 10.1080/00021369.1986.10867399

M3 - Article

AN - SCOPUS:0009743199

VL - 50

SP - 421

EP - 429

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 2

ER -