Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
The structure was elucidated in this paperNCBI PubMed ID: 23000215Publication DOI: 10.1016/j.carres.2012.05.015Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: S. Drouillard sophie.drouillard
cermav.cnrs.fr
Institutions: Centre de recherche sur Macromolecules Vegetales (CERMAV-CNRS), BP 53, F-38041 Grenoble Cedex 9, France
The α-Gal epitope is a carbohydrate structure, Galα-3Galβ-4GlcNAc-R, expressed on glycoconjuguates in many mammals, but not in humans. Species that do not express this epitope have present in their serum large amounts of natural anti-Gal antibodies, which contribute to organ hyperacute rejection during xenotransplantation. We first describe the efficient conversion of lactose into isoglobotriaose (Galα-3Galβ-4Glc) using high cell density cultures of a genetically engineered Escherichia coli strain expressing the bovine gene for α-1,3-galactosyltransferase. Attempts to produce the Galili pentasaccharide (Galα-3Galβ-4GlcNAcβ-3Galβ-4Glc) by additionally expressing the Neisseria meningitis lgtA gene for β-1,3-N-acetylglucosaminyltransferase and the Helicobacter pylori gene for β-1,4-galactosyltransferase were unsuccessful and led to the formation of a series of long chain oligosaccharides formed by the repeated addition of the trisaccharide motif [Galβ-4GlcNAcβ-3Galα-3] onto a lacto-N-neotetraose primer. The replacement of LgtA by a more specific β-1,3-N-acetylglucosaminyltransferase from H. pylori, which was unable to glycosylate α-galactosides, prevented the formation of these unwanted compounds and allowed the successful formation of the Galili pentasaccharide and longer α-Gal epitopes.
Escherichia coli, Oligosaccharides, fermentation, Metabolic engineering, α-Gal epitope, a-Gal epitope
Structure type: oligomer
Location inside paper: p.88, (8)
Contained glycoepitopes: IEDB_115013,IEDB_130645,IEDB_135813,IEDB_136044,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_138950,IEDB_141495,IEDB_141794,IEDB_141807,IEDB_142487,IEDB_142488,IEDB_146664,IEDB_149558,IEDB_151528,IEDB_151531,IEDB_190606,IEDB_742249,IEDB_918314,IEDB_983931,SB_165,SB_166,SB_173,SB_187,SB_192,SB_195,SB_6,SB_7,SB_87,SB_88
Methods: 13C NMR, 1H NMR, TLC, ESI-MS, chemical methods, genetic methods
Enzymes that release or process the structure: α-1,3-galactosyltransferase (recombinant α-3GalT), lgtA
Synthetic data: enzymatic
Comments, role: genetically engineered strain Escherichia coli
Related record ID(s): 29178, 29490, 29491, 29492, 29493, 29494, 29495, 29496, 29497, 29498, 29499, 29500, 29501, 29502, 29504, 29505, 29506, 29507, 29508, 29509, 29510, 29511, 29512, 29513, 29514
NCBI Taxonomy refs (TaxIDs): 562Reference(s) to other database(s): GTC:G92830YQ
Show glycosyltransferases
NMR conditions: in D2O at 293 K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
4,3,3,2 Ac 176.24 23.43
4,3,3 bDGlcpN 103.98 56.96 ? ? ? 62.25
4,3 aDGalp 96.87 ? 80.24 70.18 71.64 62.02
4 bDGalp 103.98 ? 78.39 66.08 ? 61.70
bDGlcp 97.01 75.01 ? 79.63 ? 61.36
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
4,3,3,2 Ac - 2.06
4,3,3 bDGlcpN 4.73 3.78 ? ? ? ?
4,3 aDGalp 5.14 3.94 4.05 4.26 4.23 ?
4 bDGalp 4.55 3.68 3.81 4.20 ? ?
bDGlcp 4.69 3.31 ? 3.71 ? ?
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
4,3,3,2 Ac 23.43/2.06
4,3,3 bDGlcpN 103.98/4.73 56.96/3.78 ?/? ?/? ?/? 62.25/?
4,3 aDGalp 96.87/5.14 ?/3.94 80.24/4.05 70.18/4.26 71.64/4.23 62.02/?
4 bDGalp 103.98/4.55 ?/3.68 78.39/3.81 66.08/4.20 ?/? 61.70/?
bDGlcp 97.01/4.69 75.01/3.31 ?/? 79.63/3.71 ?/? 61.36/?
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 |
|---|
| 4,3,3,2 | Ac |
| 2.06 | |
| 4,3,3 | bDGlcpN | 4.73 | 3.78 | ? | ? | ? | ? |
| 4,3 | aDGalp | 5.14 | 3.94 | 4.05 | 4.26 | 4.23 | ? |
| 4 | bDGalp | 4.55 | 3.68 | 3.81 | 4.20 | ? | ? |
| | bDGlcp | 4.69 | 3.31 | ? | 3.71 | ? | ? |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 |
|---|
| 4,3,3,2 | Ac | 176.24 | 23.43 | |
| 4,3,3 | bDGlcpN | 103.98 | 56.96 | ? | ? | ? | 62.25 |
| 4,3 | aDGalp | 96.87 | ? | 80.24 | 70.18 | 71.64 | 62.02 |
| 4 | bDGalp | 103.98 | ? | 78.39 | 66.08 | ? | 61.70 |
| | bDGlcp | 97.01 | 75.01 | ? | 79.63 | ? | 61.36 |
|
![](nmr_drawer.py?width=750&height=130&nucleus=C&rotate_labels=false&spectrum=[{"s":176.24,"l":"1","c":[0,0,0]},{"s":23.43,"l":"2","c":[0,0,0]},{"s":103.98,"l":"1","c":[0,0,225]},{"s":56.96,"l":"2","c":[0,0,225]},{"s":62.25,"l":"6","c":[0,0,225]},{"s":96.87,"l":"1","c":[230,0,30]},{"s":80.24,"l":"3","c":[230,0,30]},{"s":70.18,"l":"4","c":[230,0,30]},{"s":71.64,"l":"5","c":[230,0,30]},{"s":62.02,"l":"6","c":[230,0,30]},{"s":103.98,"l":"1","c":[0,165,0]},{"s":78.39,"l":"3","c":[0,165,0]},{"s":66.08,"l":"4","c":[0,165,0]},{"s":61.7,"l":"6","c":[0,165,0]},{"s":97.01,"l":"1","c":[120,0,225]},{"s":75.01,"l":"2","c":[120,0,225]},{"s":79.63,"l":"4","c":[120,0,225]},{"s":61.36,"l":"6","c":[120,0,225]}])
The spectrum also has 8 signals at unknown positions (not plotted). |
There is only one chemically distinct structure:
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Found 1 record.
Displayed record 1
