The cell envelope of Gram-positive bacteria is decorated with a variety of polysaccharides. In this study wall teichoic acid (WTA) and neutral polysaccharides were isolated from the cell envelope of bovine mastitis Streptococcus uberis. The polysaccharides were released by lysozyme treatment, and purified by hydrophobic interaction chromatography. Further separation was achieved utilizing anion-exchange chromatography which yielded two products, that is, a neutral polysaccharide with a high content of Rha and less Glc (rhamnan) and an anionic phosphate-rich one containing glycerol and Glc (WTA). The structures of these molecules were elucidated applying 1D and 2D nuclear magnetic resonance experiments as well as chemical analyses. In the rhamnan sample two independent molecules were identified, that is, a glucorhamnan with the structure →2)-α-L-Rhap-(1→3)-[α-D-Glcp-(1→2)-]α-L-Rhap-(1→, and a homopolymeric rhamnan →2)-α-L-Rhap-(1→3)-α-L-Rhap-(1→. The WTA comprised a polyphosphoglycerol chain substituted nonstoichiometrically with β-Glcp.

NMR analysis, rhamnan, WTA structure, mastitis, Streptococcus uberis, SCWPs

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
0,3,0,2	%bDGlcp	103.3	74.4	76.7	70.8	76.9	61.9
0,3,0	x?Gro	66.1	78.5	66.1
0,3	P
0,3,0,3,0	x?Gro	67.8	71.6	63.2
0,3,0,3	P
0,2	%bDGlcp
0	x?Gro
	P


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
0,3,0,2	%bDGlcp	4.47	3.31	3.51	3.40	3.46	3.77-3.90
0,3,0	x?Gro	4.03-4.11	4.22	4.03-4.11
0,3	P
0,3,0,3,0	x?Gro	3.90-3.96	3.91	3.60-3.68
0,3,0,3	P
0,2	%bDGlcp
0	x?Gro
	P


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
0,3,0,2	%bDGlcp	103.3/4.47	74.4/3.31	76.7/3.51	70.8/3.40	76.9/3.46	61.9/3.77-3.90
0,3,0	x?Gro	66.1/4.03-4.11	78.5/4.22	66.1/4.03-4.11
0,3	P
0,3,0,3,0	x?Gro	67.8/3.90-3.96	71.6/3.91	63.2/3.60-3.68
0,3,0,3	P
0,2	%bDGlcp
0	x?Gro
	P

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

Warning = sub-repeat ending at xXP?: unknown unit count was assumed to be 3
O=P(O)(O)OCC(COP(=O)(O)OCC(COP(=O)(O)OCC(COP(=O)(O)OCC(COP(=O)(O)OCC(O)CO)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O

1436.869 g/mol (C₃₉H₇₇O₄₆P₅)