title: Plate assay for endo-beta-N-acetylglucosaminidase activity using a chromogenic substrate synthesized by transglycosylation with Arthrobacter endo-beta-N-acetylglucosaminidase.
abstract: The transglycosylation activity of Arthrobacter endo-beta-N-acetylglucosaminidase (Endo-A) was used for the enzymatic synthesis of a novel oligosaccharide, Man6GlcNAc-5-bromo-4-chloro-3-indolyl-beta-glucoside (Man6GlcNAc-Glc-beta-X). Various endo-beta-N-acetylglucosaminidases hydrolyzed this oligosaccharide, producing Man6GlcNAc and Glc-beta-X. The E. coli strains coexpressing Endo-A and beta-glucosidase formed blue colonies in the presence of Man6GlcNAc-Glc-beta-X. Therefore, endo-beta-N-acetylglucosaminidase activity could be directly detected by the plate assay. This simple plate assay is useful for screening microorganisms for endo-beta-N-acetylglucosaminidase activity.
authors.affiliation: Department of Life Sciences, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Fujita
authors.forename: Y
authors.initials: Y
authors.lastname: Asada
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: K
authors.initials: K
authors.lastname: Takegawa