Effect of sucrose in culture media on the location of glucosyltransferase of Streptococcus mutans and cell adherence to glass surfaces

Abstract

Streptococcus mutans strain B13 (serotype d) almost exclusively produced free glucosyltransferase (GTase) in the culture supernatant when grown in sucrosefree TTY broth medium, which was composed of Trypticase (Baltimore Biological Laboratory [BBL] Cockeysville, Md.), tryptose (Difco Laboratories, Detroit, Mich.), yeast extract (BBL), salts, and 1% glucose. Organisms grown in sucrosefree TTY broth retained very weak cell-associated GTase activity and did not adhere significantly to glass surfaces in the presence of exogenous sucrose. If sucrose was added to TTY broth, however, GTase was found on the cell surface where cell-bound, water-insoluble glucans were synthesized. Most commercially available products of Todd-Hewitt broth were found to contain trace amounts of sucrose, as did Trypticase soy broth (BBL), whereas brain heart infusion broth (Difco and BBL) was found to be essentially free of sucrose. Almost all detectable GTase activity was cell associated when S. mutans B13 was grown in Todd-Hewitt or trypticase soy broth. Heat-treated B13 cells grown in Todd-Hewitt broth and cell-free, water-insoluble glucans bound free GTase and produced marked adherence in the presence of sucrose. Experiments strongly suggest that the binding sites for free GTase are the surface glucans, and cell-associated and extracellular GTases are most likely alternate states of the same enzyme protein. Malmo, Sweden). Culture media. To prepare extracellular GTase of S. mutans, TH broths (Difco Laboratories, Detroit, Mich., and Baltimore Biological Laboratory [BBL], Cockeysville, Md.), and Trypticase soy (TS) broth (BBL), tryptic soy (TS) broth (Difco), and BHI broths (BBL and Difco) were used. A broth medium desig-

Serotype‐Dependent Inhibition of Glucan Synthesis and Cell Adherence of Streptococcus mutans by Antibody against Glucosyltransferase of Serotype e S. mutans

“…It is of interest that antibody directed against extracellular GTase significantly inhibited the cell-associated GTase activity which resulted in the marked reduction in adherence to a glass surface of viable S. mutans cells ( Table 2). It has been found that a portion of the extracellular GTase of S. mutans will bind to the surface of S. mutans cells (18,20,25,28,29), and therefore significant inhibition of GTase activity and cell adherence can be expected.…”

Relationship of the chemical structure and immunobiological activities of lipoteichoic acid from Streptococcus faecalis (Enterococcus hirae) ATCC 9790

“…S. faecalis (Enterococcus hirae) ATCC 9790 was grown aerobically at 37 °C for 6 h in trypticase-tryptose-yeast, extract medium [10]. Cells were harvested by centrifugation and delipidated with 0.1 M sodium acetate buffer (pH 4.5)choloroform-methanol (1 : 1 : 2, v/v).…”

Polysaccharide production by cell free transferases in saliva in relation to salivary microflora

“…In spite of this variability, the significant correlations found between specific organisms and the polysaccharide production indicate that the floral composition may be one determining factor of the salivary enzyme activity level. The transferase activity of in vitro grown organisms is strongly influenced by ecologic factors such as growth rate and substrate availabihty (17,20). ROLLA et al (2) observed marked effects of sucrose rinses on the salivary transferase activity.…”