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Volume 131, Issue 12

Purification and Properties of Extracellular Glucosyltransferase Synthesizing 1,6-, 1,3-α-D-Glucan from Streptococcus mutans Serotype a Free

Abstract

Summary: An extracellular glucosyltransferase (sucrose: 1,6-, 1,3-α-D-glucan 3-α- and 6-α-D-glucosyltransferase, EC 2.4.1.-) of HS6 (serotype ) was purified from culture supernatant by DEAE-Sepharose chromatography and preparative isoelectric focusing. The molecular weight measured by SDS-PAGE was 159000 and the isoelectric point was pH 4.9. The specific activity was 89.7 i.u. (mg protein) and the optimum pH was 6.0. The value for sucrose was 4.9 mM and the enzyme activity was not stimulated by exogenous dextran T10. Glucan was synthesized from sucrose by the purified enzyme and consisted of 49.1 mol% 1,6-α-linked glucose and 33.9 mol% 1,3-α-linked glucose, with 13.6 mol% terminal glucose and 3.3 mol% 1,3,6-α-branched glucose.

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© Society for General Microbiology 1985
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1985-12-01
2024-04-20
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References

  1. Carlsson J. 1970; A levansucrase from Streptococcus mutans . Caries Research 4:97–113
     [Google Scholar]
  2. 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]
  3. Ciardi J. E., Beaman A. J., Wittenberger C. L. 1977; Purification, resolution, and interaction of the glucosyltransferases of Streptococcus mutans 6715.. Injection and Immunity 18:237–246
     [Google Scholar]
  4. Côté G. L., Robyt J. F. 1982; Isolation and partial characterization of an extracellular glucansucrase from Leuconostoc mesenteroides NRRL B-1355 that synthesizes an alternating (1→6), (1→3)-α-d-glucan.. Carbohydrate Research 101:57–74
     [Google Scholar]
  5. Dubois M. L., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugars and related substances.. Analytical Chemistry 28:350–356
     [Google Scholar]
  6. Ebisu S., Kato K., Kotani S., Misaki A. 1975; Structural differences in fructans elaborated by Streptococcus mutans and Strep. salivarius . Journal of Biochemistry 78:879–887
     [Google Scholar]
  7. 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]
  8. Fukui K., Fukui Y., Moriyama T. 1974; Some immunochemical properties of dextransucrase and invertase from Streptococcus mutans . Infection and Immunity 10:985–990
     [Google Scholar]
  9. 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]
  10. Fukui K., Kokeguchi S., Kato K., Miyake Y., Nogami R., Moriyama T. 1983; Immunochemical properties of glucosyltransferases from Streptococcus mutans . Infection and Immunity 39:762–766
     [Google Scholar]
  11. Fukushima K., Motoda R., Takada K., Ikeda T. 1981; Resolution of Streptococcus mutans glucosyltransferases into two components essential to water-insoluble glucan synthesis.. FEBS Letters 128:213–216
     [Google Scholar]
  12. Gabriel O., Wang S. -F. 1969; Determination of enzymatic activity in polyacrylamide gels. I. Enzymes catalyzing the conversion of nonreducing substrates to reducing products.. Analytical Biochemistry 27:545–554
     [Google Scholar]
  13. 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]
  14. Hakomori S. 1964; A rapid permethylation of glycolipid, and polysaccharide catalyzed by methylsulfinyl carbanion in dimethyl sulfoxide.. Journal of Biochemistry 55:205–208
     [Google Scholar]
  15. Koga T., Sato S., Yakushiji T., Inoue M. 1983; Separation of insoluble and soluble glucan-synthesizing glucosyltransferases of Streptococcus mutans OMZ176 (serotype d).. FEMS Microbiology Letters 16:127–130
     [Google Scholar]
  16. Linzer M., Slade H. D. 1974; Purification and characterization of Streptococcus mutans group d cell wall polysaccharide antigen.. Infection and Immunity 10:361–368
     [Google Scholar]
  17. 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]
  18. Mukasa H., Slade H. D. 1973b; Mechanism of adherence of Streptococcus mutans to smooth surfaces. I. Roles of insoluble dextran-levan synthetase enzymes and cell wall polysaccharide antigen in plaque formation.. Infection and Immunity 8:555–562
     [Google Scholar]
  19. Mukasa H., Slade H. D. 1974a; Mechanism of adherence of Streptococcus mutans to smooth surfaces. II. Nature of the binding site and the adsorption of dextran-levan synthetase enzymes on the cell-wall surface of the Streptococcus.. Infection and Immunity 9:419–429
     [Google Scholar]
  20. Mukasa H., Slade H. D. 1974b; Mechanism of the adherence of Streptococcus mutans to smooth surfaces. III. Purification and properties of the enzyme complex responsible for adherence.. Infection and Immunity 10:1135–1145
     [Google Scholar]
  21. 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]
  22. 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]
  23. 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]
  24. Ouchterlony O. 1958; Diffusion in gel methods for immunological analysis.. Progress in Allergy 5:1–9
     [Google Scholar]
  25. Russell R. R. B. 1979; Use of Triton X-100 to overcome the inhibition of fructosyltransferase by SDS.. Analytical Biochemistry 97:173–175
     [Google Scholar]
  26. Scales W. R., Long L. W., Edwards J. R. 1975; Purification and characterization of a glycosyltransferase complex from the culture broth of Streptococcus mutans FA 1.. Carbohydrate Research 42:325–338
     [Google Scholar]
  27. Shimamura A., Tsumori H., Mukasa H. 1982; Purification and properties of Streptococcus mutans extracellular glucosyltransferase.. Biochimica et biophysica acta 702:72–80
     [Google Scholar]
  28. Shimamura A., Tsumori H., Mukasa H. 1983; Three kinds of extracellular glucosyltransferases from Streptococcus mutans 6715 (serotype g).. FEBS Letters 157:79–84
     [Google Scholar]
  29. Somogyi M. 1945; A new reagent for the determination of sugars.. Journal of Biological Chemistry 160:61–68
     [Google Scholar]
  30. Tsumori H., Shimamura A., Mukasa H. 1983a; Purification and properties of extracellular glucosyltransferases from Streptococcus mutans serotype a . Journal of General Microbiology 129:3251–3259
     [Google Scholar]
  31. Tsumori H., Shimamura A., Mukasa H. 1983b; Comparative study of Streptococcus mutans extracellular glycosyltransferases by isoelectric focusing.. Journal of General Microbiology 129:3261–3269
     [Google Scholar]
  32. Tsumori H., Shimamura A., Mukasa H. 1985; Purification and properties of extracellular glucosyltransferase synthesizing 1,3-α-d-glucan from Streptococcus mutans serotype a . Journal of General Microbiology 131:553–559
     [Google Scholar]
  33. Vesterberg O. 1971; Isoelectric focusing of proteins.. Methods in Enzymology 22:389–412
     [Google Scholar]
  34. Vesterberg O., Hansen L., Sjösten A. 1977; Staining of proteins after isoelectric focusing in gels by new procedures.. Biochimica et biophysica acta 491:160–166
     [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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Purification and Properties of Extracellular Glucosyltransferase Synthesizing 1,6-, 1,3-α-D-Glucan from Streptococcus mutans Serotype a
Microbiology 131, 3347 (1985); https://doi.org/10.1099/00221287-131-12-3347
/content/journal/micro/10.1099/00221287-131-12-3347
/content/journal/micro/10.1099/00221287-131-12-3347
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