Plate assay for endo-beta-N-acetylglucosaminidase activity using a chromogenic substrate synthesized by transglycosylation with Arthrobacter endo-beta-N-acetylglucosaminidase.
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.
Fujita
K
K
Department of Life Sciences, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan.
Asada
Y
Y
Yamamoto
K
K
Takegawa
K
K