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 journalArticlepeer-review

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
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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