Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Host organism: Ictalurus punctatus
Associated disease: motile aeromonad septicemia (MAS)
The structure was elucidated in this paperNCBI PubMed ID: 31683071Publication DOI: 10.1016/j.carres.2019.107858Journal NLM ID: 0043535Publisher: Elsevier
Correspondence: C. Heiss <cheiss
uga.edu>
Institutions: Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, Athens, GA, 30602, USA, Department of Biological Sciences, Auburn University, 120 West Samford Avenue, Auburn, AL, 36849, USA, Department of Biological Chemistry, University of Michigan, 1150 W. Medical Center Drive, Ann Arbor, MI, 48109, USA
A hypervirulent A. hydrophila (vAh) pathotype has been identified as the etiologic agent responsible for disease outbreaks in farmed carp species and channel catfish (Ictalurus punctatus) in China and the Southeastern United States, respectively. The possible route of infection has previously been unknown; however, virulence is believed to be multifactorial, involving the production/secretion of several virulence factors, including a high molecular weight group 4 capsular polysaccharide. Here we present chemical structural evidence of a novel capsule- and LPS-associated O-antigen found present in vAh isolated during these disease outbreaks. In this study, the chemical structure of the vAh O-antigen was determined by chemical analysis, Smith degradation, mass spectrometry, and 2D proton and carbon nuclear magnetic resonance (NMR) spectroscopy and found to be unique among described bacterial O-antigens. The O-antigen consists of hexasaccharide repeating units featuring a 4)-α-l-Fucp-(1-3)-β-d-GlcpNAc-(1-4)-α-l-Fucp-(1-4)-β-d-Glcp-(1- backbone, substituted with single residue side chains of α-d-Glcp and α-d-Quip3NAc linked to O-3 of the two fucose residues. The polysaccharide is partially O-acetylated on O-6 of the 4-substituted β-Glcp residue.
Lipopolysaccharide, NMR, polysaccharide structure, Aeromonas hydrophila, Fish pathogen, O-antigen capsule
Structure type: polymer chemical repeating unit
Location inside paper: fig.1B, table 3, OPS-D
Compound class: O-antigen
Contained glycoepitopes: IEDB_135813,IEDB_136045,IEDB_137340,IEDB_141807,IEDB_142488,IEDB_142489,IEDB_144562,IEDB_144998,IEDB_145669,IEDB_146664,IEDB_150092,IEDB_151531,IEDB_152214,IEDB_174333,IEDB_983931,SB_192,SB_86
Methods: 13C NMR, 1H NMR, gel filtration, NMR-2D, GC-MS, mild acid hydrolysis, GC, Smith degradation, MALDI-TOF MS, de-O-acetylation, composition analysis, SEC, HILIC
Comments, role: de-O-acetylated OPS (OPS-D).
Related record ID(s): 767, 768, 769, 770, 1026
NCBI Taxonomy refs (TaxIDs): 1288394Reference(s) to other database(s): GTC:G64964JD
Show glycosyltransferases
NMR conditions: in D2O at 323 K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
4,4,3,3 aDGlcp 102.0 75.0 74.4 71.6 74.4 63.2
4,4,3 aLFucp 102.2 71.2 76.0 81.6 70.3 17.8
4,4,2 Ac ? 25.2
4,4 bDGlcpN 103.6 58.3 82.7 71.7 78.5 63.5
4,3,3 Ac ? 25.1
4,3 aDQuip3N 101.6 73.2 57.0 76.3 71.1 19.5
4 aLFucp 102.9 71.2 76.3 81.1 70.0 18.5
bDGlcp 105.5 76.7 78.6 80.8 78.9 63.6
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
4,4,3,3 aDGlcp 5.35 3.58 3.85 3.49 3.97 3.83-3.90
4,4,3 aLFucp 5.05 4.02 4.04 4.15 4.37 1.27
4,4,2 Ac - 2.03
4,4 bDGlcpN 4.74 3.89 3.77 3.52 3.48 3.77-3.91
4,3,3 Ac - 2.06
4,3 aDQuip3N 5.23 3.60 4.09 3.17 3.98 1.26
4 aLFucp 4.91 4.11 4.01 4.14 4.42 1.25
bDGlcp 4.55 3.36 3.49 3.51 3.39 3.85-4.00
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
4,4,3,3 aDGlcp 102.0/5.35 75.0/3.58 74.4/3.85 71.6/3.49 74.4/3.97 63.2/3.83-3.90
4,4,3 aLFucp 102.2/5.05 71.2/4.02 76.0/4.04 81.6/4.15 70.3/4.37 17.8/1.27
4,4,2 Ac 25.2/2.03
4,4 bDGlcpN 103.6/4.74 58.3/3.89 82.7/3.77 71.7/3.52 78.5/3.48 63.5/3.77-3.91
4,3,3 Ac 25.1/2.06
4,3 aDQuip3N 101.6/5.23 73.2/3.60 57.0/4.09 76.3/3.17 71.1/3.98 19.5/1.26
4 aLFucp 102.9/4.91 71.2/4.11 76.3/4.01 81.1/4.14 70.0/4.42 18.5/1.25
bDGlcp 105.5/4.55 76.7/3.36 78.6/3.49 80.8/3.51 78.9/3.39 63.6/3.85-4.00
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 |
|---|
| 4,4,3,3 | aDGlcp | 5.35 | 3.58 | 3.85 | 3.49 | 3.97 | 3.83
3.90 |
| 4,4,3 | aLFucp | 5.05 | 4.02 | 4.04 | 4.15 | 4.37 | 1.27 |
| 4,4,2 | Ac |
| 2.03 | |
| 4,4 | bDGlcpN | 4.74 | 3.89 | 3.77 | 3.52 | 3.48 | 3.77
3.91 |
| 4,3,3 | Ac |
| 2.06 | |
| 4,3 | aDQuip3N | 5.23 | 3.60 | 4.09 | 3.17 | 3.98 | 1.26 |
| 4 | aLFucp | 4.91 | 4.11 | 4.01 | 4.14 | 4.42 | 1.25 |
| | bDGlcp | 4.55 | 3.36 | 3.49 | 3.51 | 3.39 | 3.85
4.00 |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 |
|---|
| 4,4,3,3 | aDGlcp | 102.0 | 75.0 | 74.4 | 71.6 | 74.4 | 63.2 |
| 4,4,3 | aLFucp | 102.2 | 71.2 | 76.0 | 81.6 | 70.3 | 17.8 |
| 4,4,2 | Ac | ? | 25.2 | |
| 4,4 | bDGlcpN | 103.6 | 58.3 | 82.7 | 71.7 | 78.5 | 63.5 |
| 4,3,3 | Ac | ? | 25.1 | |
| 4,3 | aDQuip3N | 101.6 | 73.2 | 57.0 | 76.3 | 71.1 | 19.5 |
| 4 | aLFucp | 102.9 | 71.2 | 76.3 | 81.1 | 70.0 | 18.5 |
| | bDGlcp | 105.5 | 76.7 | 78.6 | 80.8 | 78.9 | 63.6 |
|
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The spectrum also has 2 signals at unknown positions (not plotted). |
There is only one chemically distinct structure:
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