Cell walls of Bacillus subtilis VKM B-760 and VKM B-764 are characterized by heterogeneous composition of teichoic acids. Polymer I with structure -6)-β-D-Galp-(1→1)-sn-Gro-(3-P-, polymer II with structure -6)-α-D-Glcp-(1→1)-sn-Gro-(3-P-, and a small amount of unsubstituted 1,3-poly(glycerol phosphate) were detected in strain VKM B-760. Strain VKM B-764 contains an analogous set of teichoic acids, but a characteristic feature of polymer II is the presence of disubstituted glycerol residue with α-glucopyranose localization in the integral chain at C-1 hydroxyl and β-glucopyranose as a side branch at C-2 hydroxyl (polymer III): -6)-α-D-Glcp-(1→1)-[β-D-Glcp-(1→2)]-sn-Gro-(3-P-. The structures of polymer I in bacilli and polymer III in Gram-positive bacteria are described for the first time. Teichoic acids were studied by chemical methods and on the basis of combined analysis of one-dimensional 1H-, 13C-, and (31)P-NMR spectra, homonuclear two-dimensional (1)H/(1)H COSY, TOCSY, and ROESY, and heteronuclear two-dimensional (1)H/(13)C gHSQC- and HMQC-TOCSY experiments. Simultaneous presence of several different structure teichoic acids in the bacillus cell walls as well as chemotaxonomical perspectives of the application of these polymers as species-specific markers for members of the Bacillus genus is discussed.

NMR spectroscopy, cell wall, teichoic acids, Bacillus subtilis

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
0,1	aDGlcp	99.9	72.9	74.4	70.5	72.2	66.1
0,2	bDGlcp	103.7	74.7	77.2	71.2	77.4	62.3
0	x?Gro	69.8	78.7	66.5
	P


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
0,1	aDGlcp	4.94	3.58	3.75	3.53	3.79	4.04-4.14
0,2	bDGlcp	4.64	3.33	3.52	3.40	3.47	3.75-3.91
0	x?Gro	3.62-3.83	4.22	4.06-4.13
	P


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
0,1	aDGlcp	99.9/4.94	72.9/3.58	74.4/3.75	70.5/3.53	72.2/3.79	66.1/4.04-4.14
0,2	bDGlcp	103.7/4.64	74.7/3.33	77.2/3.52	71.2/3.40	77.4/3.47	62.3/3.75-3.91
0	x?Gro	69.8/3.62-3.83	78.7/4.22	66.5/4.06-4.13
	P

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

*OC[C@H]1O[C@H](OCC(COP(*)(=O)O)O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2O)[C@H](O)[C@@H](O)[C@@H]1O

478.34 g/mol (C₁₅H₂₇R₂O₁₅P, R = next and previous repeats, not counted in mol weight)