Streptococcus mutans dextransucrase: availability of disaggregated enzyme after growth in a chemically defined medium

Schachtele, C F; Harlander, Susan K.; Germaine, G R

doi:10.1128/iai.13.5.1522-1524.1976

Cited by 29 publications

(33 citation statements)

References 21 publications

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“…When S. mutans was grown in sucrosefree TTY broth, significantly higher GTase activity was detected in the culture supernatant as compared with several commercial broth media, except BHI (Table 2). Other investigators (3,11,16,23) also demonstrated strong free GTase activity of S. mutans grown in media similar to TTY broth or in a chemically defined medium.…”

Section: Resultsmentioning

confidence: 86%

“…Furthermore, it is of interest to note that many "purified" GTase preparations contain a tightly bound carbohydrate (probably glucan) moiety (2,4,20). However, low-molecular-weight GTase free of contaminating glucan may be obtained by growing the bacterium in a chemically defined sucrosefree medium (23).…”

Section: Resultsmentioning

confidence: 99%

“…Synthesis of the glucans is mediated by the enzymatic action of cell-free (extracellular) and/or cell-bound glucosyltransferase(s) (GTase[s]). The location of S. mutans GTase appears to be complicated (1,3,11,16,20,22,23,28) and remains to be elucidated.…”

mentioning

confidence: 99%

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

Hamada¹,

Torii²

1978

Infect Immun

136

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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-

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Section: Resultsmentioning

confidence: 86%

Section: Resultsmentioning

confidence: 99%

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

Hamada¹,

Torii²

1978

Infect Immun

136

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“…HS6 cells grown in a sucrose-free broth were also used. It has been suggested that THB might contain small amounts of sucrose (21,24). The THB used in this experiment was found to contain 1.95 mg of glucose and 0.32 mg of sucrose per ml of broth, by gas-liquid chromatographical analysis.…”

Section: Methodsmentioning

confidence: 68%

Synthesis of Glucan on the Cell Surface of Streptococcus mutans: Chemical and Scanning Electron Microscopic Studies

Tsumori

Mukasa

Zinnaka

1982

Microbiology and Immunology

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The appearance and continuing growth of extracellular material on Streptococcus mutans HS6 cells in sucrose-containing Merthiolated buffer was observed in a scanning electron microscope and was found to be related to the glucan synthesis on the cell and to adherence of the cell to a smooth surface. Cells grown in broth completely deprived of sucrose by invertase (HS6-IV) had a characteristic, slightly rugged surface structure. On incubation of HS6-IV in the sucrosecontaining buffer, a few small globular particles appeared on the surface and grew to an irregular shape (globular to fibrilar) after several hours. The increase in the total glucan content of the cells paralleled the growth of the globular material, to which ferritin-conjugated anti-dextran globulin was found to bind. On the cell surface of cells harvested from conventional broth, both small globular and irregular structures, which possibly formed from sucrose in the broth, existed originally and continued to grow during incubation, along with the material newly appearing on the surface. The accumulation of glucan on the cells resulted in their adherence to a glass surface. The inhibition of growth of the extracellular material on the cells by trypsin, dextranase or anti-glucosyltransferase corresponded to the decrease in glucan synthesis and the loss of adhering ability. These results indicated that the material growing on the cell surface was glucan synthesized by glucosyltransferases.

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“…Schachtele et al. (21) demonstrated that the GTF enzymes from strain 6715 grown in defined medium were stimulated 11-fold by soluble primer dextrans. Comparison of GTF activity of strain GS-5 at any point during growth indicated that these enzymes are not greatly stimulated by primer (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Production of Extracellular and Cell-Associated Glucosyltransferase Activity by Streptococcus mutans During Growth on Various Carbon Sources

Janda

Kuramitsu

1978

Infect Immun

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Streptococcus mutans dextransucrase: availability of disaggregated enzyme after growth in a chemically defined medium

Cited by 29 publications

References 21 publications

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

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

Synthesis of Glucan on the Cell Surface of Streptococcus mutans: Chemical and Scanning Electron Microscopic Studies

Production of Extracellular and Cell-Associated Glucosyltransferase Activity by Streptococcus mutans During Growth on Various Carbon Sources

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