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Archives of Microbiology

, Volume 147, Issue 3, pp 207–212 | Cite as

Electrophoretic studies of extracellular glucosyltransferases and fructosyltransferases from seventeen strains of Streptococcus mutans

  • S. Kametaka
  • S. Hayashi
  • Y. Miyake
  • H. Suginaka
Original Papers

Abstract

Streptococcus mutans was classified by the electrophoretic properties of glucosyltransferases (GTases) and fructosyltransferases (FTases). The cells of serotypes a, d and g did not release extracellular FTases, although those from other serotypes did. The enzymes from cells of serotypes d and g synthesized a good deal of insoluble polysaccharide compared with other serotypes. The enzymes were applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and polyacrylamide gel-isoelectric focussing (PAG-IEF). Gels were stained for their activity and protein content. Enzymes belonging to the same serotype gave the same specific pattern on both gels. The seven serotypes could be classified into the following four groups: serotypes d and g, serotype a, serotypes c, e and f, and serotype b. The results agree well with some previous reports based on other methods. The molecular weights of three GTase bands were 156K, 146K and 135K, and of four kinds of FTase bands were 108K, 95K, 80K and 76K. The isoelectric points of main enzymes were 4.25, 4.60, 5.00, 5.55 and 5.70. Those of FTases were 4.25 and 4.60.

Key words

Streptococcus mutans Glucosyltransferase Fructosyltransferase Electrophoresis Isoelectric focussing 

Abbreviations

GTase

glucosyltransferase

FTase

fructosyltransferase

SDS-PAGE

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

PAG-IEF

polyacrylamide gel-isoelectric focussing

PAS

periodic acid-Schiff

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References

  1. Bratthall D (1970) Demonstration of five serological groups of streptococcal strains resembling Streptococcus mutans. Odont Revy 21:143–152Google Scholar
  2. Brown AT, Patterson CE (1972) Heterogeneity of Streptococcus mutans strains based on their mannitol-1-phosphate dehydrogenases: criterion for rapid classification. Infect Immun 6:422–424Google Scholar
  3. Carlsson J (1970) A levansucrase from Streptococcus mutans. Caries Res 4:97–113Google Scholar
  4. Ciardi JE, Hageage Jr GJ, Wittenberger CL (1976) Multicomponent nature of glucosyltransferase system of Streptococcus mutans. J Dent Res 55:C87-C96Google Scholar
  5. Coykendall AL (1977) Proposal to elevate subspecies of Streptococcus mutans to species stature, based on their molecular composition. Int J Syst Bacteriol 27:26–30Google Scholar
  6. Evans RT, Genco RJ (1973) Inhibition of glucosyltransferase activity by antisera to know serotypes of Streptococcus mutans. Infect Immun 7:237–241Google Scholar
  7. Fitzgerald RJ, Keyes PH (1960) Demonstration of the etiologic role of streptococci in experimental caries in the hamster. J Am Dent Assoc 61:9–19Google Scholar
  8. Fukui K, Fukui Y, Moriyama T (1974) Purification and properties of dextransucrase and invertase from Streptococcus mutans. J Bacteriol 118:796–804Google Scholar
  9. Fukui K, Kokeguchi S, Kato K, Miyaka Y, Nogami R, Moriyama T (1983) Immunochemical properties of glucosyltransferases from Streptococcus mutans. Infect Immun 39:762–766Google Scholar
  10. Guggenheim B (1970) Extracellular polysaccharides and microbial plaque. Int Dent J 20:675–678Google Scholar
  11. Hamada S, Ooshima T, Torii M, Imanishi H, Masuda N, Sobue S, Kotani S (1978) Dental caries induction in experimental animals by clinical strains of Streptococcus mutans with human dental decay. Infect Immun 11:1252–1260Google Scholar
  12. Hamada S, Tai S, Slade HD (1979) Serotype-dependent inhibition of glucan synthesis and cell adherence of Streptococcus mutans by antibody against glucosyltransferase of serotype S. mutans. Microbiol Immunol 23:61–70Google Scholar
  13. Hojo S, Higuchi M (1981) The role of insoluble glucan as a diffusion barrier in dental plaque. Jpn J Oral Biol 23:527–533Google Scholar
  14. Keyes PH (1960) The infection and transmissible nature of experimental dental caries. Arch Oral Biol 1:304–320Google Scholar
  15. Kuramitsu HK (1975) Characterization of extracellular glucosyltransferase activity of Streptococcus mutans. Infect Immun 12:738–749Google Scholar
  16. Kuramitsu HK, Ingersoll L (1976) Immunological relationship between glucosyltransferases from Streptococcus mutans serotypes. Infect Immun 14:636–644Google Scholar
  17. Laas T, Olsson I (1981) pH-Gradient development and focusing speed in thin-layer polyacrylamide gel isoelectric focusing: a comparison between Pharmalyte, Ampholine, and Servalyt. Electrophoresis 2:235–239Google Scholar
  18. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  19. Littleton NW, Kakehashi S, Fitzgerald RJ (1970) Recovery of specific “caries-inducing” streptococci from carious lesion in the teeth of children. Arch Oral Biol 15:461–463Google Scholar
  20. Loesche WJ, Rowan J, Staffon LH, Loos PJ (1975) Association of Streptococcus mutans with human dental decay. Infect Immun 11:1252–1260Google Scholar
  21. Mukasa H, Slade HD (1974) Mechanism of adherence of Streptococcus mutans to smooth surfaces. III. Purification and properties of the enzyme complex responsible for adherence. Infect Immun 10:1135–1145Google Scholar
  22. Oakley BR, Kirsch DR, Morris NR (1980) A simple ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem 105:361–363Google Scholar
  23. Osborne RM, Lamberts BL, Meyer TS, Roush AH (1976) Acrylamide gel electrophoretic studies of extracellular sucrose-metabolizing enzymes of Streptococcus mutans. J Dent Res 55:77–84Google Scholar
  24. Perch B, Kjems E, Ravn T (1974) Biochemical and serological properties of Streptococcus mutans from various human and animal sources. Acta Pathol Microbiol Scand 82:357–370Google Scholar
  25. Russell RRB (1976) Classification of Streptococcus mutans strains by SDS gel electrophoresis. Microbios Lett 2:55–59Google Scholar
  26. Russell RRB (1979) Use of Triton X-100 to overcome the inhibition of fructosyltransferase by SDS. Anal Biochem 97:173–175Google Scholar
  27. Shklair IL, Keene HJ (1976) Biochemical characterization and distribution of Streptococcus mutans in three diverse populations. In: Stiles HM, Loesche WJ, O'Brien TC (eds) Microbial aspect of dental caries. Information Retrieval Inc., Washington DC, pp 201–210Google Scholar
  28. Smith DJ, Taubman MA (1977) Antigenic relatedness of glucosyltransferase enzymes from Streptococcus mutans. Infect Immun 15:91–103Google Scholar
  29. Suginaka H, Wada H, Kotani S (1975) Initial products in utilization of sucrose by Actinomyces viscosus. Biken J 18:271–274Google Scholar
  30. Sweeley CC, Wells WW, Bentley R (1966) Gas chromatography of carbohydrates. In: Neufeld EF, Ginsburg V (eds) Methods in enzymology, vol VIII. Academic Press, New York, pp 95–97Google Scholar
  31. Tanzer JM, Brown AT, MacInerney MF, Woodiel FN (1977) Comparative study of invertases of Streptococcus mutans. Infect Immun 16:318–327Google Scholar
  32. Tsumori H, Shimamura A, Mukasa H (1979) Comparison among type-specific properties of glucosyltransferases and cell wall antigens of Streptococcus mutans. Bull Natl Def Med Col 2: 91–109Google Scholar
  33. Tsumori H, Shimamura A, Mukasa H (1983) Comparative study of Streptococcus mutans extracellular glycosyltransferases by isoelectric focusing. J Gen Microbiol 129:3261–3269Google Scholar
  34. Zacharius RM, Zell TE, Morrison JH, Wordlock JJ (1969) Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem 30:148–152Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • S. Kametaka
    • 1
  • S. Hayashi
    • 1
  • Y. Miyake
    • 2
  • H. Suginaka
    • 2
  1. 1.Research and Development DivisionRohto Pharmaceutical Co., Ltd.OsakaJapan
  2. 2.Department of Microbiology, School of DentistryHiroshima UniversityHiroshimaJapan

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