9LR5

Crystal structure of Bacteroides thetaiotaomicron GH84 O-GlcNAcase D243N mutant

Summary for 9LR5

• Entry DOI: 10.2210/pdb9lr5/pdb
• Descriptor: O-GlcNAcase BT_4395, CALCIUM ION (2 entities in total)
• Functional Keywords: glycoside hydrolase, hydrolase, glycosidase
• Biological source: Bacteroides thetaiotaomicron VPI-5482
• Total number of polymer chains: 1
• Total formula weight: 83315.29

Authors

Ohnuma, T.,Takeshita, D. (deposition date: 2025-01-29, release date: 2025-04-23, Last modification date: 2025-07-30)

Primary citation

Okuno, R.,Nakada, S.,Tonomura, K.,Aso, Y.,Takeshita, D.,Ohnuma, T.,Tanaka, T.
beta 1,6-Selective Enzymatic N-Acetylglucosamination Catalyzed by the Family GH84 N-Acetyl-beta-D-glucosaminidase from Bacteroides thetaiotaomicron and its Glycosyl Acceptor Specificity.
Chem Asian J, 20:e202500142-e202500142, 2025

PubMed Abstract: The chemoenzymatic synthesis of oligosaccharides presents a highly attractive methodology with significant potential for diverse applications, particularly through using various glycosidases. In this study, the O-glycan core 6 disaccharide moiety, GlcNAcβ1-6GalNAc, was successfully synthesized via enzymatic glycosylation using an N-acetyl-β-D-glucosaminidase from Bacteroides thetaiotaomicron (BtOGA), a member of glycoside hydrolase family 84 (GH84), alongside an N-acetyl-D-glucosamine oxazoline derivative (GlcNAc-oxa) as the glycosyl donor. Furthermore, an investigation into glycosyl acceptor recognition in BtOGA-catalyzed enzymatic glycosylation indicated that the presence of an aromatic group at the anomeric position and an axial hydroxy group at the 4-position of the saccharide moiety is crucial for effective recognition of BtOGA as a glycosyl acceptor. The protecting-group-free chemoenzymatic synthesis of the core 6 disaccharide moiety was achieved by integrating the direct synthesis of GlcNAc-oxa thorough Shoda activation method using a water-soluble dehydration condensing agent in an aqueous medium, followed by BtOGA-catalyzed enzymatic glycosylation.

PubMed: 40195893
DOI: 10.1002/asia.202500142

Experimental method

X-RAY DIFFRACTION (3.099 Å)