Secondary cell wall polysaccharides (SCWPs) are important structural components of the Bacillus cell wall and contribute to the array of antigens presented by these organisms in both spore and vegetative forms. We previously found that antisera raised to Bacillus anthracis spore preparations cross-reacted with SCWPs isolated from several strains of pathogenic B. cereus, but did not react with other phylogenetically related but nonpathogenic Bacilli, suggesting that the SCWP from B. anthracis and pathogenic B. cereus strains share specific structural features. In this study, SCWPs from three strains of B. cereus causing severe or fatal pneumonia (G9241, 03BB87 and 03BB102) were isolated and subjected to structural analysis and their structures were compared to SCWPs from B. anthracis. Complete structural analysis was performed for the B. cereus G9241 SCWP using NMR spectroscopy, mass spectrometry and derivatization methods. The analyses show that SCWPs from B. cereus G9241 has a glycosyl backbone identical to that of B. anthracis SCWP, consisting of multiple trisaccharide repeats of: →6)-α-D-GlcpNAc-(1→4)-β-D-ManpNAc-(1→4)-β-D-GlcpNAc-(1→. Both the B. anthracis and pathogenic B. cereus SCWPs are highly substituted at all GlcNAc residues with α- and β-Gal residues, however, only the SCWPs from B. cereus G9241 and 03BB87 carry an additional α-Gal substitution at O-3 of ManNAc residues, a feature lacking in the B. anthracis SCWPs. Both the B. anthracis and B. cereus SCWPs are pyruvylated, with an approximate molecular mass of approximately 12,000 Da. The implications of these findings regarding pathogenicity and cell wall structure are discussed.
13C NMR, 1H NMR, NMR-2D, GC-MS, HF solvolysis, MALDI-TOF MS, composition analysis, immunofluorescence analyses
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
4,4,6,4,4,2 Ac 176.20 20.99
4,4,6,4,4,3 aDGalp 99.40 69.76 70.19 69.76 71.70 61.6
4,4,6,4,4,4 bDGalp 103.48 71.70 73.30 69.33 76.21 61.8
4,4,6,4,4 aDGlcpN 96.61 52.79 77.30 77.70 69.98 67.6
4,4,6,4,2 Ac 175.97 22.93
4,4,6,4,3 aDGalp 101.56 68.68 70.19 69.97 72.13 62.46
4,4,6,4 bDManpN 99.40 54.51 80.28 77.27 75.56 60.7
4,4,6,2 Ac 176.20 20.99
4,4,6,3 aDGalp 98.33 69.54 70.19 69.76 71.70 61.4
4,4,6 bDGlcpN 101.54 55.15 75.98 77.31 75.99 60.7
4,4,2 Ac 174.46 22.93
4,4,3 aDGalp 99.62 69.76 70.19 69.76 71.70 61.6
4,4,4 bDGalp 103.48 71.70 73.30 69.33 76.21 61.8
4,4 aDGlcpN 98.76 53.86 71.69 79.86 71.69 67.6
4,2 Ac 175.97 22.93
4 bDManpN 100.04 54.51 73.41 76.20 75.35 61.2
2 Ac 174.46 22.93
3 aDGalp 97.91 69.54 70.19 69.76 71.70 61.4
bDGlcpN 101.54 55.15 75.98 77.31 75.99 60.7
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
4,4,6,4,4,2 Ac - 2.08
4,4,6,4,4,3 aDGalp 5.55 3.76 3.72 3.99 3.83 3.74-3.83
4,4,6,4,4,4 bDGalp 4.44 3.54 3.64 3.94 3.66 3.75-3.82
4,4,6,4,4 aDGlcpN 5.76 4.16 4.00 4.10 3.81 3.98-3.98
4,4,6,4,2 Ac - 2.03
4,4,6,4,3 aDGalp 5.05 3.77 3.65 3.93 4.15 3.73-3.73
4,4,6,4 bDManpN 4.91 4.65 4.24 4.10 3.52 3.89-3.91
4,4,6,2 Ac - 2.08
4,4,6,3 aDGalp 5.63 3.80 3.72 3.99 3.81 3.72-3.72
4,4,6 bDGlcpN 4.65 3.90 3.90 4.10 3.52 3.74-3.92
4,4,2 Ac - 2.08
4,4,3 aDGalp 5.52 3.76 3.71 3.98 3.86 3.74-3.83
4,4,4 bDGalp 4.44 3.54 3.64 3.94 3.66 3.75-3.82
4,4 aDGlcpN 5.24 3.91 3.86 3.63 3.77 3.93-3.95
4,2 Ac - 2.03
4 bDManpN 4.90 4.50 4.05 3.67 3.51 3.82-3.91
2 Ac - 2.08
3 aDGalp 5.68 3.80 3.71 3.98 3.81 3.72-3.72
bDGlcpN 4.65 3.90 3.90 4.10 3.52 3.74-3.92
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
4,4,6,4,4,2 Ac 20.99/2.08
4,4,6,4,4,3 aDGalp 99.40/5.55 69.76/3.76 70.19/3.72 69.76/3.99 71.70/3.83 61.6/3.74-3.83
4,4,6,4,4,4 bDGalp 103.48/4.44 71.70/3.54 73.30/3.64 69.33/3.94 76.21/3.66 61.8/3.75-3.82
4,4,6,4,4 aDGlcpN 96.61/5.76 52.79/4.16 77.30/4.00 77.70/4.10 69.98/3.81 67.6/3.98-3.98
4,4,6,4,2 Ac 22.93/2.03
4,4,6,4,3 aDGalp 101.56/5.05 68.68/3.77 70.19/3.65 69.97/3.93 72.13/4.15 62.46/3.73-3.73
4,4,6,4 bDManpN 99.40/4.91 54.51/4.65 80.28/4.24 77.27/4.10 75.56/3.52 60.7/3.89-3.91
4,4,6,2 Ac 20.99/2.08
4,4,6,3 aDGalp 98.33/5.63 69.54/3.80 70.19/3.72 69.76/3.99 71.70/3.81 61.4/3.72-3.72
4,4,6 bDGlcpN 101.54/4.65 55.15/3.90 75.98/3.90 77.31/4.10 75.99/3.52 60.7/3.74-3.92
4,4,2 Ac 22.93/2.08
4,4,3 aDGalp 99.62/5.52 69.76/3.76 70.19/3.71 69.76/3.98 71.70/3.86 61.6/3.74-3.83
4,4,4 bDGalp 103.48/4.44 71.70/3.54 73.30/3.64 69.33/3.94 76.21/3.66 61.8/3.75-3.82
4,4 aDGlcpN 98.76/5.24 53.86/3.91 71.69/3.86 79.86/3.63 71.69/3.77 67.6/3.93-3.95
4,2 Ac 22.93/2.03
4 bDManpN 100.04/4.90 54.51/4.50 73.41/4.05 76.20/3.67 75.35/3.51 61.2/3.82-3.91
2 Ac 22.93/2.08
3 aDGalp 97.91/5.68 69.54/3.80 70.19/3.71 69.76/3.98 71.70/3.81 61.4/3.72-3.72
bDGlcpN 101.54/4.65 55.15/3.90 75.98/3.90 77.31/4.10 75.99/3.52 60.7/3.74-3.92
report error
Found 1 record.
Displayed record 1
]; septicemia [ICD11: MA15.Y 
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