Structure and functions of linkage unit intermediates in the biosynthesis of ribitol teichoic acids in Staphylococcus aureus H and Bacillus subtilis W23

The stepwise formation and characterization of linkage unit intermediates and their functions in ribitol teichoic acid biosynthesis were studied with membranes obtained from Staphylococcus aureus H and Bacillus subtilis W23. The formation of labeled polymer from CDP‐[ 14 C]ribitol and CDP‐glycerol in each membrane system was markedly stimulated by the addition of N ‐acetylmannosaminyl(β1→4) N ‐acetylglucosamine (ManNAc‐GlcNAc) linked to pyrophosphorylpolyisoprenol. Whereas incubation of S. aureus membranes with CDP‐glycerol and ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol led to synthesis of (glycerol phosphate) 1–3 ‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol, incubation of B. subtilis membranes with the same substrates yielded (glycerol phosphate) 1–2 ‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol. In S. aureus membranes, (glycerol phosphate) 2 ‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol as well as (glycerol phosphate) 3 ‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol served as an acceptor for ribitol phosphate units, but (glycerol phosphate)‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol did not. In B. subtilis W23 membranes, (glycerol phosphate)‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol served as a better acceptor for ribitol phosphate units than (glycerol phosphate) 2 ‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol. In this membrane system (ribitol phosphate)‐(glycerol phosphate)‐ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol was formed from ManNAc‐[ 14 C]GlcNAc‐ P P ‐prenol, CDP‐glycerol and CDP‐ribitol. The results indicate that (glycerol phosphate) 1–3 ‐ManNAc‐GlcNAc‐ P P ‐prenol and (glycerol phosphate) 1–2 ‐ManNAc‐GlcNAc‐ P P ‐prenol are involved in the pathway for the synthesis of wall ribitol teichoic acids in S. aureus H and B. subtilis W23 respectively.