Structural studies on the linkage unit between poly(galactosylglycerol phosphate) and peptidoglycan in cell walls of Bacillus coagulans.

Structural studies were carried out on the linkage units in the teichoic-acid--glycopeptide complexes isolated from lysozyme digests of the cell walls of Bacillus coagulans AHU 1366. On treatment with 47% hydrogen fluoride, the complexes gave a disaccharide characterized as glucosyl(beta 1----4)N-acetylglucosamine together with major fragments, galactosyl(alpha 1----2)glycerol. By means of Smith degradation and partial acid hydrolysis, the teichoic acid chain was shown to be composed of the repeating units, galactosyl(alpha 1----2)glycerol-3(1)-phosphate, which were joined by phosphodiester bonds at C-6 of the galactose residues. The mild alkaline hydrolysis of teichoic-acid-linked glycan fragments yielded teichoic acid chains and disaccharide-linked glycan fragments, from which the disaccharide, glucosyl(beta 1----4)N-acetylglucosamine, was liberated by mild acid hydrolysis, whereas the same disaccharide linked to the teichoic acid chain was obtained by direct heating of the cell walls at pH 2.5. In addition, the presence of specialized glycerol phosphate units in the linkage region was shown by the isolation of tris(glycerol phosphate)3-glucosyl(beta 1----4)N-acetylglucosamine from the products of the Smith degradation of the teichoic-acid--glycopeptide complexes. Thus, it is concluded that the poly(galactosylglycerol phosphate) chain in the cell walls of B. coagulans AHU 1366 is linked to peptidoglycan through a novel linkage unit, bis(glycerol phosphate)-3-glucosyl(beta 1----4)N-acetylglucosamine.