Purification and characterization of Streptococcus mutans group d cell wall polysaccharide antigen

Abstract

The Streptococcus mutans group d antigen of strain B13 has been purified and characterized with respect to chemical composition and immunochemical properties. The antigen was extracted from lyophilized cells or cell walls by using 5% trichloroacetic acid at 5 C for 16 h. The antigen could also be extracted with water or 0.01 N HCl at 100 C for 20 min. The antigen was purified by ion-exchange and gel chromatography and was found to contain 96% carbohydrate, 1.6% protein, and 0.3% phosphorus. Characterization by gas chromatography indicated that the polysaccharide was composed of galactose and glucose in a 2:1 ratio. The antigen contained two serologically active sites: one site specific for group d and a second site common to both group d and group a strains. Agar diffusion and immunoelectrophoresis indicated that the two sites existed on a single molecule. The immunological specificity of the group d polysaccharide site depended on a terminal d-galactose. The purified B13 antigen did not react with antisera specific for the glycerol teichoic acid from streptococci. Anti-d serum rapidly agglutinated whole cells, indicating that the antibody receptor sites of the polysaccharide antigen were at the surface of the streptococcal cell.