Plate assay for endo-β-N-acetylglucosaminidase activity using a chromogenic substrate synthesized by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase

Kiyotaka Fujita, Yasuhiko Asada, Kenji Yamamoto, Kaoru Takegawa

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The transglycosylation activity of Arthrobacter endo-β-N-acetylglucosaminidase (Endo-A) was used for the enzymatic synthesis of a novel oligosaccharide, Man6GlcNAc-5-bromo-4-chloro-3-indolyl-β-glucoside (Man6GlcNAc-Glc-β-X). Various endo-β-N-acetylglucosaminidases hydrolyzed this oligosaccharide, producing Man6GlcNAc and Glc-β-X. The E. coli strains coexpressing Endo-A and β-glucosidase formed blue colonies in the presence of Man6GlcNAc-Glc-β-X. Therefore, endo-β-N-acetylglucosaminidase activity could be directly detected by the plate assay. This simple plate assay is useful for screening microorganisms for endo-β-N-acetylglucosaminidase activity.

Original languageEnglish
Pages (from-to)462-464
Number of pages3
JournalJournal of Bioscience and Bioengineering
Volume90
Issue number4
DOIs
Publication statusPublished - Jan 1 2000

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Chromogenics
Arthrobacter
Chromogenic Compounds
Acetylglucosaminidase
Oligosaccharides
Assays
Substrates
Microorganisms
Escherichia coli
Screening
Glucosidases
Glucosides
endo-alpha-sialidase

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Plate assay for endo-β-N-acetylglucosaminidase activity using a chromogenic substrate synthesized by transglycosylation with Arthrobacter endo-β-N-acetylglucosaminidase. / Fujita, Kiyotaka; Asada, Yasuhiko; Yamamoto, Kenji; Takegawa, Kaoru.

In: Journal of Bioscience and Bioengineering, Vol. 90, No. 4, 01.01.2000, p. 462-464.

Research output: Contribution to journalArticle

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abstract = "The transglycosylation activity of Arthrobacter endo-β-N-acetylglucosaminidase (Endo-A) was used for the enzymatic synthesis of a novel oligosaccharide, Man6GlcNAc-5-bromo-4-chloro-3-indolyl-β-glucoside (Man6GlcNAc-Glc-β-X). Various endo-β-N-acetylglucosaminidases hydrolyzed this oligosaccharide, producing Man6GlcNAc and Glc-β-X. The E. coli strains coexpressing Endo-A and β-glucosidase formed blue colonies in the presence of Man6GlcNAc-Glc-β-X. Therefore, endo-β-N-acetylglucosaminidase activity could be directly detected by the plate assay. This simple plate assay is useful for screening microorganisms for endo-β-N-acetylglucosaminidase activity.",
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AB - The transglycosylation activity of Arthrobacter endo-β-N-acetylglucosaminidase (Endo-A) was used for the enzymatic synthesis of a novel oligosaccharide, Man6GlcNAc-5-bromo-4-chloro-3-indolyl-β-glucoside (Man6GlcNAc-Glc-β-X). Various endo-β-N-acetylglucosaminidases hydrolyzed this oligosaccharide, producing Man6GlcNAc and Glc-β-X. The E. coli strains coexpressing Endo-A and β-glucosidase formed blue colonies in the presence of Man6GlcNAc-Glc-β-X. Therefore, endo-β-N-acetylglucosaminidase activity could be directly detected by the plate assay. This simple plate assay is useful for screening microorganisms for endo-β-N-acetylglucosaminidase activity.

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