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

Authors
Fujita, K Asada, Y Yamamoto, K Takegawa, K
Citation
K. Fujita et al., Plate assay for endo-beta-N-acetylglucosaminidase activity using a chromogenic substrate synthesized by transglycosylation with Arthrobacter endo-beta-N-acetylglucosaminidase, J BIOSCI BI, 90(4), 2000, pp. 462-464
Citations number
15
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
ISSN journal
13891723 → ACNP
Volume
90
Issue
4
Year of publication
2000
Pages
462 - 464
Database
ISI
SICI code
1389-1723(200010)90:4<462:PAFEAU>2.0.ZU;2-R
Abstract
The transglycosylation activity of Arthrobacter endo-beta -N-acetylglucosam inidase (Endo-A) was used for the enzymatic synthesis of a novel oligosacch aride, Man(6)GlcNAc-5-bromo-4-chloro-3-indolyl-beta -glucoside (Man(6)GlcNA c-Glc-beta -X). Various endo-beta -N-acetylglucosaminidases hydrolyzed this oligosaccharide, producing Man(6)GlcNAc and Glc-beta -X. The E. coli strai ns coexpressing Endo-A and beta -glucosidase formed blue colonies in the pr esence of Man(6)GlcNAc-Glc-beta -X. Therefore, endo-beta -N-acetylglucosami nidase activity could be directly detected by the plate assay. This simple plate assay is useful for screening microorganisms for endo-beta -N-acetylg lucosaminidase activity.