Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Klebsiella pneumoniae [ICD11:
XN741 
]
The structure was elucidated in this paperNCBI PubMed ID: 34973762Publication DOI: 10.1016/j.carbpol.2021.118944Journal NLM ID: 8307156Publisher: Elsevier
Correspondence: Shih-Hsiung Wu <shwu
gate.sinica.edu.tw>
Institutions: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan, Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, Department of Physics, Chemistry, and Biology, Linkoping University, Sweden, Department of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan, Doctoral Degree Program in Marine Biotechnology, National Sun Yat-sen University, Kaohsiung, Taiwan, Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan
Klebsiella pneumoniae serotype KN2 is a carbapenem-resistant strain and leads to the health care-associated infections, such as bloodstream infections. Its capsular polysaccharide (CPS) was isolated and cleaved by a specific enzyme from a bacteriophage into a hexasaccharide-repeating unit. With GC-MS, NMR, and Mass analyses, the structure of KN2 CPS was determined to be {→3)-β-D-Glcp-(1→3)-[α-D-GlcpA-(1→4)-β-D-Glcp-(1→6)]-α-D-Galp-(1→6)-β-D-Galp-(1→3)-β-D-Galp-(1→}n. We demonstrated that 1 μg/mL CPS could stimulate J774A.1 murine macrophages to release tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in vitro. Also, we proved that KN2 CPS induced the immune response through Toll-like receptor 4 (TLR4) in the human embryonic kidney (HEK)-293 cells. Strikingly, the hexasaccharide alone shows the same immune response as the CPS, suggesting that the hexasaccharide can shape the adaptive immunity to be a potential vaccine adjuvant. The glucuronic acid (GlcA) on other polysaccharides can affect the immune response, but the GlcA-reduced KN2 CPS and hexasaccharide still maintain their immunomodulatory activities.
NMR, capsular polysaccharide, Toll-like receptor 4, bacteriophage glycosidase, K.pneumoniae
Structure type: polymer chemical repeating unit ; >1000000
Location inside paper: abstract, Fig. 3(e), 3(f)
Compound class: CPS
Contained glycoepitopes: IEDB_115136,IEDB_134624,IEDB_136044,IEDB_136906,IEDB_137472,IEDB_140630,IEDB_141794,IEDB_142488,IEDB_146664,IEDB_151528,IEDB_190606,IEDB_742248,IEDB_983931,SB_163,SB_165,SB_166,SB_187,SB_192,SB_195,SB_7,SB_88
Methods: 13C NMR, 1H NMR, NMR-2D, GC-MS, sugar analysis, ESI-MS, carboxyl reduction, cytokine production, enzymatic activity assay, Quanti-Blue assay, Ciucanu/Kerek methylation, Ms/MS
Related record ID(s): 21055
NCBI Taxonomy refs (TaxIDs): 573
Show glycosyltransferases
There is only one chemically distinct structure:
Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Klebsiella pneumoniae [ICD11:
XN741 ]
The structure was elucidated in this paperNCBI PubMed ID: 34973762Publication DOI: 10.1016/j.carbpol.2021.118944Journal NLM ID: 8307156Publisher: Elsevier
Correspondence: Shih-Hsiung Wu <shwu
gate.sinica.edu.tw>
Institutions: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan, Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, Department of Physics, Chemistry, and Biology, Linkoping University, Sweden, Department of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan, Doctoral Degree Program in Marine Biotechnology, National Sun Yat-sen University, Kaohsiung, Taiwan, Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan
Klebsiella pneumoniae serotype KN2 is a carbapenem-resistant strain and leads to the health care-associated infections, such as bloodstream infections. Its capsular polysaccharide (CPS) was isolated and cleaved by a specific enzyme from a bacteriophage into a hexasaccharide-repeating unit. With GC-MS, NMR, and Mass analyses, the structure of KN2 CPS was determined to be {→3)-β-D-Glcp-(1→3)-[α-D-GlcpA-(1→4)-β-D-Glcp-(1→6)]-α-D-Galp-(1→6)-β-D-Galp-(1→3)-β-D-Galp-(1→}n. We demonstrated that 1 μg/mL CPS could stimulate J774A.1 murine macrophages to release tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in vitro. Also, we proved that KN2 CPS induced the immune response through Toll-like receptor 4 (TLR4) in the human embryonic kidney (HEK)-293 cells. Strikingly, the hexasaccharide alone shows the same immune response as the CPS, suggesting that the hexasaccharide can shape the adaptive immunity to be a potential vaccine adjuvant. The glucuronic acid (GlcA) on other polysaccharides can affect the immune response, but the GlcA-reduced KN2 CPS and hexasaccharide still maintain their immunomodulatory activities.
NMR, capsular polysaccharide, Toll-like receptor 4, bacteriophage glycosidase, K.pneumoniae
Structure type: oligomer ; 1027 [M+Na]+
Location inside paper: Fig. 1, Fig. 3(e), 3(f), table 2
Trivial name: repeat unit
Compound class: CPS
Contained glycoepitopes: IEDB_115136,IEDB_134624,IEDB_136044,IEDB_136906,IEDB_137472,IEDB_140630,IEDB_141794,IEDB_142488,IEDB_146664,IEDB_151528,IEDB_190606,IEDB_742248,IEDB_983931,SB_163,SB_165,SB_166,SB_187,SB_192,SB_195,SB_7,SB_88
Methods: 13C NMR, 1H NMR, NMR-2D, GC-MS, sugar analysis, ESI-MS, carboxyl reduction, cytokine production, enzymatic activity assay, Quanti-Blue assay, Ciucanu/Kerek methylation, Ms/MS
Comments, role: The oligosaccharide obtained by treatment of the KN2 CPS with KN2 enzyme; NMR -3)bDGalp residue, 1H-1/C-1: 4.63/98.8 ? ? ? ? ?. Published NMR chemical shift of C5 #3,6,6,4_aDGlcpA (79.1) seemed erroneous.
Related record ID(s): 8498
NCBI Taxonomy refs (TaxIDs): 573
Show glycosyltransferases
NMR conditions: in D2O at 298 K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
3,6,3 bDGlcp 106.4 75.9 78.7 74.4 75.3 63.5
3,6,6,4 aDGlcpA 101.1 74.5 75.1 72.4 ? 178.0
3,6,6 bDGlcp 105.3 75.9 78.4 78.6 71.9 62.7
3,6 aDGalp 101.2 69.7 82.3 71.6 72.6 72.0
3 bDGalp 106.8 73.7 75.2 69.8 73.6 69.5
aDGalp 94.9 70.1 81.7 71.9 72.8 63.8
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
3,6,3 bDGlcp 4.67 3.44 3.77 3.68 3.70 3.73-3.92
3,6,6,4 aDGlcpA 5.46 3.51 3.69 3.40 3.79 -
3,6,6 bDGlcp 4.50 3.30 3.49 3.40 3.71 3.79-3.79
3,6 aDGalp 5.01 3.99 3.91 4.23 4.11 3.86-4.04
3 bDGalp 4.65 3.61 3.69 3.99 3.66 3.72-3.91
aDGalp 5.28 3.98 3.99 4.22 4.09 3.70-3.73
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
3,6,3 bDGlcp 106.4/4.67 75.9/3.44 78.7/3.77 74.4/3.68 75.3/3.70 63.5/3.73-3.92
3,6,6,4 aDGlcpA 101.1/5.46 74.5/3.51 75.1/3.69 72.4/3.40 ?/3.79
3,6,6 bDGlcp 105.3/4.50 75.9/3.30 78.4/3.49 78.6/3.40 71.9/3.71 62.7/3.79-3.79
3,6 aDGalp 101.2/5.01 69.7/3.99 82.3/3.91 71.6/4.23 72.6/4.11 72.0/3.86-4.04
3 bDGalp 106.8/4.65 73.7/3.61 75.2/3.69 69.8/3.99 73.6/3.66 69.5/3.72-3.91
aDGalp 94.9/5.28 70.1/3.98 81.7/3.99 71.9/4.22 72.8/4.09 63.8/3.70-3.73
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 |
|---|
| 3,6,3 | bDGlcp | 4.67 | 3.44 | 3.77 | 3.68 | 3.70 | 3.73
3.92 |
| 3,6,6,4 | aDGlcpA | 5.46 | 3.51 | 3.69 | 3.40 | 3.79 |
|
| 3,6,6 | bDGlcp | 4.50 | 3.30 | 3.49 | 3.40 | 3.71 | 3.79
3.79 |
| 3,6 | aDGalp | 5.01 | 3.99 | 3.91 | 4.23 | 4.11 | 3.86
4.04 |
| 3 | bDGalp | 4.65 | 3.61 | 3.69 | 3.99 | 3.66 | 3.72
3.91 |
| | aDGalp | 5.28 | 3.98 | 3.99 | 4.22 | 4.09 | 3.70
3.73 |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 |
|---|
| 3,6,3 | bDGlcp | 106.4 | 75.9 | 78.7 | 74.4 | 75.3 | 63.5 |
| 3,6,6,4 | aDGlcpA | 101.1 | 74.5 | 75.1 | 72.4 | ? | 178.0 |
| 3,6,6 | bDGlcp | 105.3 | 75.9 | 78.4 | 78.6 | 71.9 | 62.7 |
| 3,6 | aDGalp | 101.2 | 69.7 | 82.3 | 71.6 | 72.6 | 72.0 |
| 3 | bDGalp | 106.8 | 73.7 | 75.2 | 69.8 | 73.6 | 69.5 |
| | aDGalp | 94.9 | 70.1 | 81.7 | 71.9 | 72.8 | 63.8 |
|
The spectrum also has 1 signal at unknown position (not plotted). |
There is only one chemically distinct structure:
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