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	bDGalp	104.8	72.2	73.2	69.6	74.9	65.7
0	x?Gro	72.0	70.8	67.8
	P


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
0,1	bDGalp	4.43	3.56	3.67	3.99	3.85	4.05-4.11
0	x?Gro	3.72-4.01	4.03	3.91-3.99
	P


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
0,1	bDGalp	104.8/4.43	72.2/3.56	73.2/3.67	69.6/3.99	74.9/3.85	65.7/4.05-4.11
0	x?Gro	72.0/3.72-4.01	70.8/4.03	67.8/3.91-3.99
	P

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

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

316.199 g/mol (C₉H₁₇R₂O₁₀P, R = next and previous repeats, not counted in mol weight)