1887

Abstract

Extracellular glucosyltransferases (sucrose: 1,6-α--glucan 3-α-and 6-α-glucosyltransferase) of HS6 (serotype ) were purified from the culture supernatant by DEAE-Sepharose chromatography, ConA-Sepharose chromatography and chromatofocusing. The enzymes I and II with specific activities of 6·20 and 5·86 i.u. mg, respectively, exhibited slightly different isoelectric points (pI 4·5 and 4·2) and the molecular weights were estimated to be 161000 and 174000, respectively, by SDS-PAGE. The enzymes had the same optimum pH of 5·5 and the same values of 1·3 m for sucrose and of 83 μ-glucose equivalent for dextran T10. By double immunodiffusion test on agar, these enzymes were immunologically identical to each other. Analysis by GLC of the glucans synthesized from sucrose by the enzymes (I and II) established that they were 1,6-α--glucans with 20 and 24·5 mol% 1,3,6-branch points, respectively. Both are therefore Afunctional enzymes.

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1983-10-01
2024-04-23
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References

  1. Carlsson J. 1970; A levansucrase from streptococcus mutans. Caries Research 4:97–113
    [Google Scholar]
  2. Chaiet L., Kempf A.J., Harman R., Kackzka E., Weston R., Nollstadt K., Wolf F.J. 1970; Isolation of a pure dextranase from Penicillium funiculosum. Applied Microbiology 20:421–426
    [Google Scholar]
  3. Chludzinski A.M., Germaine G.R., Shachatele C.F. 1976; Streptococcus mutansdextransu- crase: purification, properties, and requirement for primer dextran. Journal of Dental Research 55: (Special Issue C) C75–C86
    [Google Scholar]
  4. Ciardi J.E., Hageage G.J.Jr Wittenberger C.L. 1976; Multicomponent nature of the glucosyltransferase system of Streptococcus mutans. Journal of Dental Research 55: (Special Issue C) C87–C96
    [Google Scholar]
  5. Dubois M.L., Gilles K.A., Hamilton J.K., Rebers P.A., Smith F. 1956; Colorimetric methods for determination of sugars and related substances. Analytical Chemistry 28:350–356
    [Google Scholar]
  6. Eisenthal R., Cornish-Bowden A. 1974; The direct linear plot.A new graphical procedure for estimating enzyme kinetic parameters. Biochemical Journal 139:715–720
    [Google Scholar]
  7. Figures W.R., Edwards J.R. 1979; Resolution of the glycosyltransferase activities from two strains of Streptococcus mutans by polyacrylamide gel electrophoresis in the presence of Tween 80. Biochimica et biophysica acta 577:142–146
    [Google Scholar]
  8. Fujiwara S., Kobayashi S., Nakayama H. 1978; Development of a minimal medium for Streptococcus mutans. Archives of Oral Biology 23:601–602
    [Google Scholar]
  9. Fukui K., Fukui Y., Moriyama T. 1974a; Purification and properties of dextransucrase and invertase from Streptococcus mutans. Journal of Bacteriology 118:796–804
    [Google Scholar]
  10. Fukui K., Fukui Y., Moriyama T. 1974b; Some immunochemical properties of dextransucrase and invertase from Streptococcus mutans. Infection and Immunity 10:985–990
    [Google Scholar]
  11. Fukui K., Moriyama T., Miyake Y., Mizutani K., Tanaka O. 1982; Purification and properties of glucosyltransferase responsible for water-insoluble glucan synthesis from Streptococcus mutans. Infection and Immunity 37:1–9
    [Google Scholar]
  12. Fukushima K., Motoda R., Takada K., Ikeda T. 1981; Resolution of Streptococcus mutansglucosyltransferases into two components essential to water-insoluble glucan synthesis. FEBS Letters 128:213–216
    [Google Scholar]
  13. Germaine G.R., Harlander S.K., Leung W.L.S., Schachtele C.F. 1977; Streptococcus mutansdextransucrase: functioning of primer dextran and endogenous dextranase in water-soluble and water-insoluble glucan synthesis. In fection and Immunity 16:637–648
    [Google Scholar]
  14. Gibbons R.J., Nygaard M. 1968; Synthesis of insoluble dextran and its significance in the formation of gelatinous deposits by plaque-forming streptococci. Archives of Oral Biology 13:1249–1262
    [Google Scholar]
  15. Hakomori S. 1964; A rapid permethylation of glycolipid, and polysaccharide catalyzed by methylsulfinylcarbanion in dimethyl sulfoxide. Journal of Biochemistry 55:205–208
    [Google Scholar]
  16. Hare M.D., Svensson S., Walker G.J. 1978; Characterization of the extracellular, water-insoluble a-D-glucans of oral streptococci by methylation analysis, and by enzymic synthesis and degradation. Carbohydrate Research 66:245–264
    [Google Scholar]
  17. Kapitany R.A., Zebrowski E.J. 1973; A high resolution PAS stain for polyacrylamide gel electrophoresis. Analytical Biochemistry 56:361–369
    [Google Scholar]
  18. Meyer T.S., Lamberts B.L., Egan R.S. 1978; Structural studies of extracellular glucans of Streptococcus mutans by proton magnetic resonance. Carbohydrate Research 66:33–42
    [Google Scholar]
  19. Mukasa H., Slade H.D. 1973a; Extraction, purification and chemical and immunological properties of the Streptococcus mutans group a polysaccharide cell wall antigen. Infection and Immunity 8:190–198
    [Google Scholar]
  20. Mukasa H., Slade H.D. 1973b; Mechanism of adherence of Streptococcus mutans to smooth surfaces. I. Roles of insoluble dextran-levansynthetase enzymes and cell wall polysaccharide antigen in plaque formation. In fection and Immunity 8:555–562
    [Google Scholar]
  21. Mukasa H., Slade H.D. 1974; Mechanism of adherence of Streptococcus mutans to smooth surfaces. III. Purification and properties of the enzyme complex responsible for adherence. In fection and Immunity 10:1135–1145
    [Google Scholar]
  22. Mukasa H., Shimamura A., Tsumori H. 1979; Effect of salts on water-insoluble glucan formation by glucosyltransferase of Streptococcus mutans. Infection and Immunity 23:564–570
    [Google Scholar]
  23. Mukasa H., Shimamura A., Tsumori H. 1982a; Direct activity stains for glycosidase and glucosyltransferase after isoelectric focusing in horizontal polyacrylamide gel layers. Analytical Biochemistry 123:276–284
    [Google Scholar]
  24. Mukasa H., Shimamura A., Tsumori H. 1982b; Purification and characterization of basic glucosyltransferase from Streptococcus mutans serotype c. Biochimica et biophysica acta 719:81–89
    [Google Scholar]
  25. Ouchterlony O. 1958; Diffusion in gel methods for immunological analysis. Progress in Allergy 5:1–9
    [Google Scholar]
  26. Russell R.R.B. 1979a; Glycosyltransferases of Streptococcus mutans strain Ingbritt. Microbios 23:135–146
    [Google Scholar]
  27. Russell R.R.B. 1979b; Use of Triton X-100 to overcome the inhibition of fructosyltransferase by SDS. Analytical Biochemistry 91:173–175
    [Google Scholar]
  28. Schachtele C.F., Staat R.H., Harlander S.K. 1975; Dextranases from oral bacteria: inhibition of water-insoluble glucan production and adherence to smooth surfaces by Streptococcus mutans. Infection and Immunity 12:309–317
    [Google Scholar]
  29. Shimamura A., Tsumori H., Mukasa H. 1982; Purification and properties of Streptococcus mutansextracellular glucosyltransferase. Biochimica et biophysica acta 702:72–80
    [Google Scholar]
  30. Smith D.J., Taubman M.A. 1977; Antigenic relatedness of glucosyltransferase enzymes from Streptococcus mutans. Infection and Immunity 15:91–103
    [Google Scholar]
  31. Somogyi M. 1945; A new reagent for the determination of sugars. Journal of Biological Chemistry 160:61–73
    [Google Scholar]
  32. Tsumori H., Mukasa H., Zinnaka Y. 1982; Synthesis of glucan on the cell surface of Streptococcus mutans: chemical and scanning electron microscopic studies. Microbiology and Immunology 26:677–688
    [Google Scholar]
  33. Usui T., Sugiyama H., Seto S., Araya S., Nisizawa T., Imai S., Kosaka K. 1975; Structural determination of glucans from Streptococcus mutansJC-2 (dental caries bacterium) by carbon-13 nuclear magnetic resonance. Journal of Biochemistry 78:225–227
    [Google Scholar]
  34. Walker G.J., Hare M.D. 1977; Metabolism of the polysaccharides of human dental plaque. Part II. Purification and properties of Cladosporium resinae(l-3)-α-d-glucanase, and the enzymic hydrolysis of glucans synthesized by extracellular D-glucosyltransferases of oral streptococci. Carbohydrate Research 58:415–432
    [Google Scholar]
  35. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
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