1887

Abstract

The four recombinant glucosyltransferases (GTFs), GtfJ, GtfK, GtfL and GtfM, that had previously been cloned from ATCC 25975, were individually expressed in and their glucan products and kinetic properties were analysed. GtfJ was a primer-dependent GTF which synthesized an insoluble glucan composed mainly of α-(13)-linked glucosyl residues in the presence of dextran T-10. GtfK was primer-stimulated, and produced a linear soluble dextran without any detectable branch points both in the absence and in the presence of dextran T-10. GtfL was primer-independent and produced a mixed-linkage insoluble glucan composed of approximately equal proportions of α-(13)-and α-(16)-linked glucosyl residues. GtfL was inhibited by dextran T-10. GtfM was primer-independent and produced a soluble dextran with approximately 5% α-(13)-linked glucosyl residues. GtfM was essentially unaffected by the presence of dextran T-10. The results confirmed that each enzyme represented one of the four possible combinations of primer-dependency and product solubility and that each possessed unique biosynthetic properties. The soluble dextrans formed by GtfK and GtfM, as well as the mixed-linkage insoluble glucan formed by GtfL, were also capable of acting as primers for the primer-dependent GtfJ and the primer-stimulated GtfK. Unexpectedly, the linear dextran produced by GtfK was by far the least effective either at priming itself or at activating and priming the primer-dependent GtfJ.

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1995-06-01
2021-05-12
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References

  1. Carlsson J. 1969; A numerical taxonomic study of human oral streptococci.. Odont Revy 19:137–159
    [Google Scholar]
  2. Cheetham N.W.H., Walker G.J., Pearce B.J., Fiala-Beer E., Taylor C. 1991; Structures of water-soluble a-D-glucans synthesized from sucrose by glucosyltransferases isolated from Streptococcus sobrinus culture filtrates.. Carbohydr Polym 14:3–16
    [Google Scholar]
  3. Colson P., Jennings H.J., Smith I.C.P. 1974; Composition, sequence and conformation of polymers and oligomers of glucose as revealed by carbon 13 nuclear magnetic resonance.. J Am Chem Soc 96:8081–8087
    [Google Scholar]
  4. Drucker D.B., Shakespeare A.P., Green R.M. 1984; The production of dental plaque and caries by the bacterium Streptococcus salivarius in gnotobiotic WAG/RIJ rats.. Arch Oral Biol 29:437–443
    [Google Scholar]
  5. Eifuku H., Yoshimitsu-Narita A., Sato S., Yakushui T., Inoue M. 1989; Production and partial characterization of the extracellular polysaccharides from oral Streptococcus salivarius. . Carbohydr Res 194:247–260
    [Google Scholar]
  6. 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. . Infect Immun 37:1–9
    [Google Scholar]
  7. Giffard P.M., Simpson C.L., Milward C.P., Jacques N.A. 1991; Molecular characterization of a cluster of at least two glucosyltransferase genes in Streptococcus salivarius ATCC 25975.. J Gen Microbiol 137:2577–2593
    [Google Scholar]
  8. Giffard P.M., Allen D.M., Milward C.P., Simpson C.L., Jacques N.A. 1993; Sequence of the gtfK gene of Streptococcus salivarius ATCC 25975 and evolution of the gtf genes of oral streptococci.. J Gen Microbiol 139:1511–1522
    [Google Scholar]
  9. Gilmore K.S., Russell R.R.B., Ferretti J.J. 1993; Expression of gtfS is essential for normal insoluble glucan synthesis by Streptococcus downei. . Infect Immun 61:1246–1250
    [Google Scholar]
  10. Gilpin M.L, Russell R.R.B., Morrissey P. 1985; Cloning and expression of two Streptococcus mutans glucosyltransferases in Escherichia coli K12.. Infect Immun 49:414–416
    [Google Scholar]
  11. Gorin P.J.A. 1981; Carbon-13 nuclear magnetic resonance spectroscopy of polysaccharides.. Adv Carbohydr Chem Biochem 38:13–104
    [Google Scholar]
  12. Gough J.A., Murray N.E. 1983; Sequence diversity among related genes for recognition of specific targets in DNA molecules.. J Mol Biol 166:1–19
    [Google Scholar]
  13. Hanada N., Kuramitsu H.K. 1989; Isolation and characterization of the Streptococcus mutans gtfD gene, coding for primer-dependent soluble glucan synthesis.. Infect Immun 57:2079–2085
    [Google Scholar]
  14. Hanada N., Takehara T. 1987a; Comparison of different water- soluble glucan synthases from Streptococcus mutans serotype g. . Microbios 50:147–152
    [Google Scholar]
  15. Hanada N., Takehara T. 1987b; (1 → 3)-ɑ-D-glucan synthase from Streptococcus mutans AHT (serotype g) does not synthesise glucan without primer.. Carbohydr Res 168:120–124
    [Google Scholar]
  16. Hanada N., Takehara T. 1991; Substrate specificity of hydrolase activity of the primer-dependent glucosyltransferases from Streptococcus sobrinus. . Microbios 66:21–25
    [Google Scholar]
  17. Hanada N., Katayama T., Kunimori A., Yamashita Y., Takehara T. 1993; Four different types of glucans synthesised by glucosyltransferases from Streptococcus sobrinus. . Microbios 73:23–35
    [Google Scholar]
  18. Hare M.D., Svensson S., Walker G.J. 1978; Characterization of the extracellular, water-insoluble ɑ-D-glucans of oral streptococci by methylation analysis, and by enzymic synthesis and degradation.. Carbohydr Res 66:254–264
    [Google Scholar]
  19. Jacques N.A. 1983; Membrane perturbation by cerulenin modulates glucosyltransferase secretion and acetate uptake by Streptococcus salivarius. . J Gen Microbiol 129:3293–3302
    [Google Scholar]
  20. McCabe M.M. 1985; Purification and characterization of a primer-independent glucosyltransferase from Streptococcus mutans 6715-13 mutant 27.. Infect Immun 50:771–777
    [Google Scholar]
  21. Mayer R.M., Matthews M.M., Futerman C.L., Parnaik V.K., Jung S.M. 1981; Dextransucrase: acceptor substrate reactions.. Arch Biochem Biophys 208:278–287
    [Google Scholar]
  22. Miller J.H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. Milnes A.R., Bowden G. H., Gates D., Tate R. 1993a; Predominant cultivable microorganisms on the tongue of preschool children.. Microb Ecol Health Dis 6:229–235
    [Google Scholar]
  24. Milnes A. R., Bowden G.H., Gates D., Tate R. 1993b; Normal microbiota on the teeth of preschool children.. Microb Ecol Health Dis 6:213–227
    [Google Scholar]
  25. Munro C., Michalek S.M., Macrina F.L. 1991; Cariogenicity of Streptococcus mutans V403 glucosyltransferase and fructosyl- transferase mutants constructed by allellic exchange.. Infect Immun 59:2316–2323
    [Google Scholar]
  26. Murray N.E., Brammar W.H., Murray K. 1977; Lambdoid phages that simplify the recovery of in vitro recombinants.. Mol & Gen Genet 150:53–61
    [Google Scholar]
  27. Nakano Y.J., Kuramitsu H.K. 1992; Mechanism of Streptococcus mutans glucosyltransferases: hybrid-enzyme analysis.. J Bacteriol 174:5639–5646
    [Google Scholar]
  28. Pitty LJ., Giffard P.M., Gilpin M.L., Russell R.R.B., Jacques N.A. 1989; Cloning and expression of glycosyltransferase activities from Streptococcus salivarius. . J Dent Ris 68:1681–1682
    [Google Scholar]
  29. Russell R.R.B., Gilpin M.L., Mukasa H., Dougan G. 1987; Characterization of glucosyltransferase expressed from a Streptococcus sobrinus gene cloned in Escherichia coli. . J Gen Microbiol 133:935–944
    [Google Scholar]
  30. Russell R.R.B., Gilpin M.L, Hanada N., Yamashita Y., Shibata Y., Takehara T. 1990; Characterization of the product of the gtfS gene of Streptococcus downei a primer-independent enzyme synthesizing oligo-isomaltosaccharides.. J Gen Microbiol 136:1631–1637
    [Google Scholar]
  31. Sato S., Inoue M. 1991; Partial characterization of the glucosyltransferases of an oral Streptococcus salivarius strain.. Microbios 68:179–188
    [Google Scholar]
  32. Shimamura A. 1989; Use of 13C-N. M. R. spectroscopy for the quantitative estimation of 3-0- and 3,6-di-O-substituted D- glucopyranosyl residues in a-D-glucans formed by the d- glucosyltransferases of Streptococcus sobrinus. . Carbohydr Res 185:239–248
    [Google Scholar]
  33. Shimamura A., Tsumori H., Mukasa H. 1982; Purification and properties of Streptococcus mutans extracellular glucosyltransferase.. Biochim Biophys Acta 702:72–80
    [Google Scholar]
  34. Shimamura A., Tsumori H., Mukasa H. 1983; Three kinds of extracellular glucosyltransferases from Streptococcus mutans 6715 (serotype g).. FEBS Lett 157:79–84
    [Google Scholar]
  35. Shimamura A., Nakano Y.J., Mukasa H., Kuramitsu H.K. 1994; Identification of amino acid residues in Streptococcus mutans glucosyltransferases influencing the structure of the glucan product.. J Bacterial 176:4845–4850
    [Google Scholar]
  36. Simpson C.L, Giffard P. M., Jacques N. A. 1995; Streptococcus salivarius ATCC 25975 possesses at least two genes coding for primer-independent glucosyltransferases.. Infect Immun 63:609–621
    [Google Scholar]
  37. Takehara T., Hanada N., Saeki E. 1984; Interaction of glucosyltransferase isozymes on glucan synthesis by Streptococcus mutans AHT (serotype g).. Microbios Lett 27:113–120
    [Google Scholar]
  38. Takehara T., Ansai T., Yamashita Y., Itoh-Andoh M., Hanada N., Kunimori A. 1992; Mechanism of water-insoluble glucan synthesis in Streptococcus sobrinus. . Oral Microbiol Immunol 7:155–158
    [Google Scholar]
  39. Taylor C., Cheetham N. W. H., Slodki M. E., Walker G. J. 1990; Action of endo-(1→6)-α-glucanases on the soluble dextrans produced by three extracellular α-D-glucosyltransferases of Streptococcus sobrinus. . Carbobydr Polymers 13: 423–436
    [Google Scholar]
  40. Walker G.J. 1973; Preparation of isomaltose oligosaccharides labelled with 14C in the non-reducing terminal unit, and their use in studies of dextranase activity.. J Dent Res 52:1–10
    [Google Scholar]
  41. Walker G.J. 1978; Dextrans.. Int Rev Biochem 16:75–126
    [Google Scholar]
  42. Walker G.J., Jacques N.A. 1987; Polysaccharides of oral streptococci.. In Sugar Transport and Metabolism in Gram-positive Bacteria pp. 39–68 Reizer J., Peterkofsky A. Edited by Chichester: Ellis Horwood;
    [Google Scholar]
  43. Walker G.J., Schuerch C. 1986; Activity of branched dextrans in the acceptor reaction of a glucosyltransferase (GTF-I) from Streptococcus mutans OMZ176.. Carbohydr Res 146:259–270
    [Google Scholar]
  44. Walker G.J., Cheetham N.W.H., Taylor C. 1990; Productivity of four α-D-glucosyltransferases released by Streptococcus sobrinus under defined conditions in continuous culture.. Carbohydr Polym 13:399–421
    [Google Scholar]
  45. Wittenberger C.L, Beaman A. J., Lee L. N. 1978; Tween 80 effect on glucosyltransferase synthesis by Streptococcus salivarius. . J Bacteriol 133:231–239
    [Google Scholar]
  46. Yamashita Y., Hanada N., Takehara T. 1988a; A novel glucosyltransferase from Streptococcus mutans produces oligo- isomaltosaccharides.. Biochem Biophys Res Commun 150:687–693
    [Google Scholar]
  47. Yamashita Y., Shigeoka T., Hanada N., Takehara T. 1988b; Immunological properties of the primer-independent glucosyltransferase of Streptococcus mutans serotypes d and g. . J Gen Microbiol 134:1223–1227
    [Google Scholar]
  48. Yamashita Y., Hanada N., Takehara T. 1989; Purification of a fourth glucosyltransferase from Streptococcus sobrinus. . J Bacteriol 171:6265–6270
    [Google Scholar]
  49. Yamashita Y., Bowen W.H., Burne R.A., Kuramitsu H.K. 1993; Role of the Streptococcus mutans gtf genes in caries induction in the specific-pathogen-free rat model.. Infect Immun 61:3811–3817
    [Google Scholar]
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