Purification and Characterization of Endo-β-N-acetylglucosaminidase from a Flavobacterium sp

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

doi:10.1271/bbb1961.50.421

Purification and Characterization of Endo-β-N-acetylglucosaminidase from a Flavobacterium sp

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

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

A gram negative bacterium isolated from soil was found to produce a high level of Endo-β-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 ribonuclease B and a yeast invertase.

Original language	English
Pages (from-to)	421-429
Number of pages	9
Journal	Agricultural and Biological Chemistry
Volume	50
Issue number	2
DOIs	https://doi.org/10.1271/bbb1961.50.421
Publication status	Published - 1986
Externally published	Yes

All Science Journal Classification (ASJC) codes

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

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title = "Purification and Characterization of Endo-β-N-acetylglucosaminidase from a Flavobacterium sp",

abstract = "A gram negative bacterium isolated from soil was found to produce a high level of Endo-β-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 ribonuclease B and a yeast invertase.",

author = "Kenji Yamamoto and Setsu Kadowaki and Kaoru Takegawa and Hidehiko Kumagai and Tatsurokuro Tochikura",

year = "1986",

doi = "10.1271/bbb1961.50.421",

language = "English",

volume = "50",

pages = "421--429",

journal = "Agricultural and Biological Chemistry",

issn = "0002-1369",

publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",

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T1 - Purification and Characterization of Endo-β-N-acetylglucosaminidase from a Flavobacterium sp

AU - Yamamoto, Kenji

AU - Kadowaki, Setsu

AU - Takegawa, Kaoru

AU - Kumagai, Hidehiko

AU - Tochikura, Tatsurokuro

PY - 1986

Y1 - 1986

N2 - A gram negative bacterium isolated from soil was found to produce a high level of Endo-β-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 ribonuclease B and a yeast invertase.

AB - A gram negative bacterium isolated from soil was found to produce a high level of Endo-β-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 ribonuclease B and a yeast invertase.

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DO - 10.1271/bbb1961.50.421

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JO - Agricultural and Biological Chemistry

JF - Agricultural and Biological Chemistry

IS - 2

ER -

Purification and Characterization of Endo-β-N-acetylglucosaminidase from a Flavobacterium sp

Abstract

All Science Journal Classification (ASJC) codes

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