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Taxonomic group: bacteria / Proteobacteria (Phylum: Proteobacteria)
Host organism: Homo sapiens
Associated disease: acute haemorrhagic diarrhea [ICD11: 1A03

, ICD11: 1A03.3

]; hemolytic-uremic syndrome (HUS) [ICD11: 3A21.2

]; infection due to Escherichia coli [ICD11: XN6P4

]

NCBI PubMed ID: 26703456
Publication DOI: 10.1093/glycob/cwv169
Journal NLM ID: 9104124
Publisher: IRL Press at Oxford University Press
Correspondence: lli22

gsu.edu (L. Li); pwang11

gsu.edu (P.G. Wang)
Institutions: College of Life Science, Hebei Normal University, Shijiazhuang, Hebei 050024, China, State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 30071, China, Department of Chemistry and Center for Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA

Colitose, also known as 3,6-dideoxy-l-galactose or 3-deoxy-l-fucose, is one of the only five naturally occurred 3,6-dideoxyhexoses. Colitose was found in lipopolysaccharide of a number of infectious bacteria, including Escherichia coli O55 & O111 and Vibrio cholera O22 & O139. To date, no colitosyltransferase (ColT) has been characterized, probably due to the inaccessibility of the sugar donor, GDP-colitose. In this study, starting with chemically prepared colitose, 94.6 mg of GDP-colitose was prepared via a facile and efficient one-pot two-enzyme system involving an l-fucokinase/GDP-l-Fuc pyrophosphorylase and an inorganic pyrophosphatase (EcPpA). WbgN, a putative ColT from E. coli O55:H5 was then cloned, overexpressed, purified and biochemically characterized by using GDP-colitose as a sugar donor. Activity assay and structural identification of the synthetic product clearly demonstrated that wbgN encodes an α1,2-ColT. Biophysical study showed that WbgN does not require metal ion, and is highly active at pH 7.5-9.0. In addition, acceptor specificity study indicated that WbgN exclusively recognize lacto-N-biose (Gal β1,3-GlcNAc). Most interestingly, it was found that WbgN exhibits similar activity toward GDP-l-Fuc (kcat/Km = 9.2 min-1 mM-1) as that toward GDP-colitose (kcat/Km = 12 min-1 mM-1). Finally, taking advantage of this, type 1 H-antigen was successfully synthesized in preparative scale.

synthesis, colitosyltransferase, Escherichia coli O55, GDP-colitose, H-antigen

Structure type: suggested polymer biological repeating unit
Location inside paper: fig.2
Compound class: O-polysaccharide
Contained glycoepitopes: IEDB_130648,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_137473,IEDB_1391962,IEDB_141794,IEDB_141807,IEDB_142078,IEDB_143794,IEDB_144990,IEDB_150899,IEDB_151528,IEDB_151531,IEDB_167071,IEDB_190606,SB_137,SB_156,SB_165,SB_166,SB_173,SB_187,SB_195,SB_29,SB_7,SB_88

Methods: 13C NMR, 1H NMR, SDS-PAGE, kinetics assays, TLC, Western blotting, biophysical methods, genetic methods, biochemical methods, enzyme assay, enzymatic synthesis, chemoenzymatic synthesis, HR-ESI-MS
Biological activity: biochemically characterization of WbgN colitosyltransferase
Biosynthesis and genetic data: cloning wbgN gene

Related record ID(s): 5554, 7239, 8618, 20658, 28398, 30374, 108629, 113595, 113759
NCBI Taxonomy refs (TaxIDs): 244320
Reference(s) to other database(s): GTC:G96674TM
Show glycosyltransferases

There are 2 chemically distinct structures. Please, select:

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

SVG MW SMILES 3D mol Ionize