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Taxonomic group: bacteria / Proteobacteria (Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11: XN6P4

]

NCBI PubMed ID: 18487334
Publication DOI: 10.1128/JB.00160-08
Journal NLM ID: 2985120R
Publisher: American Society for Microbiology
Correspondence: fenglu63

nankai.edu.cn
Institutions: Department of Medicine, Department of Biochemistry, Queen's University, Kingston Ontario K7L 3N6, Canada, TEDA School of Biological Sciences and Biotechnology, Nankai University, Hongda Street, TEDA, Tianjin 300457, P.R.China, Tianjin Key Laboratory of Microbial Functional Genomics, P. R. China, Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, P. R. China, Department of Chemistry, Queen's University, Kingston Ontario K7L 3N6, Canada

The O antigens of outer membrane-bound lipopolysaccharides (LPS) in Gram-negative bacteria are oligosaccharides consisting of repeating units with various structures and antigenicities. The O56 and O152 antigens of Escherichia coli both contain a Glc-β 1-3-GlcNAc linkage within the repeating unit. We have cloned and identified the genes (wfaP in O56 and wfgD in O152) within the two O antigen gene clusters that encode glucosyltransferases involved in the synthesis of this linkage. A synthetic substrate analog of the natural acceptor substrate undecaprenol-pyrophosphate-lipid (GlcNAc-PP-PhU) was used as an acceptor and UDP-Glc as a donor substrate to demonstrate that both wfgD and wfaP encode glucosyltransferases. Enzyme products from both glucosyltransferases were isolated by HPLC and analyzed by NMR. The spectra showed the expected Glc-β 1-3-GlcNAc linkage in the products, confirming that both WfaP and WfgD are UDP-Glc: GlcNAc-pyrophosphate-lipid β-1,3-glucosyltransferases. Both WfaP and WfgD have a DxD sequence which is proposed to interact with phosphate groups of the nucleotide donor through the coordination of a metal cation, and a short hydrophobic sequence at the C terminus that may help to associate the enzymes with the inner membrane. We showed that the enzymes have similar properties and substrate recognition. They both require divalent cation (Mn(2+) or Mg(2+)) for activity, are deactivated by detergents, have a broad pH optimum, and require the pyrophosphate-sugar linkage in the acceptor substrate for full activity. Substrates lacking phosphate or pyrophosphate linked to GlcNAc were inactive. The length of the aliphatic chain of acceptor substrates also contributes to the activity.

O-antigen, gene cluster, Glucosyltransferases, enzymatic syntheses, Escherichia coli O56, Escherichia coli O152

Structure type: suggested polymer biological repeating unit
Location inside paper: p.4923, fig.1
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_135813,IEDB_136794,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_146100,IEDB_146664,IEDB_149174,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_983931,SB_170,SB_171,SB_172,SB_192,SB_7,SB_84

Methods: 13C NMR, 1H NMR, genetic methods, biochemical methods, HPLC
Biosynthesis and genetic data: genetic data
Synthetic data: enzymatic

Related record ID(s): 8382, 20438, 20659, 22850, 22851, 22852, 25691, 108697, 124618
NCBI Taxonomy refs (TaxIDs): 562
Reference(s) to other database(s): GTC:G32692RT, GlycomeDB:6888
Show glycosyltransferases

There is only one chemically distinct structure:

*O[C@@H]([C@@H]1O[C@@](O[C@H]2[C@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O)[C@@H](CO)O[C@@H](*)[C@@H]3NC(C)=O)[C@@H]2O[C@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)(C(=O)O)C[C@H](O)[C@H]1NC(C)=O)[C@H](O)CO

SVG MW SMILES 3D mol Ionize