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.
Similar content being viewed by others
Abbreviations
- GTase:
-
glucosyltransferase
- FTase:
-
fructosyltransferase
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- PAG-IEF:
-
polyacrylamide gel-isoelectric focussing
- PAS:
-
periodic acid-Schiff
References
Bratthall D (1970) Demonstration of five serological groups of streptococcal strains resembling Streptococcus mutans. Odont Revy 21:143–152
Brown AT, Patterson CE (1972) Heterogeneity of Streptococcus mutans strains based on their mannitol-1-phosphate dehydrogenases: criterion for rapid classification. Infect Immun 6:422–424
Carlsson J (1970) A levansucrase from Streptococcus mutans. Caries Res 4:97–113
Ciardi JE, Hageage Jr GJ, Wittenberger CL (1976) Multicomponent nature of glucosyltransferase system of Streptococcus mutans. J Dent Res 55:C87-C96
Coykendall AL (1977) Proposal to elevate subspecies of Streptococcus mutans to species stature, based on their molecular composition. Int J Syst Bacteriol 27:26–30
Evans RT, Genco RJ (1973) Inhibition of glucosyltransferase activity by antisera to know serotypes of Streptococcus mutans. Infect Immun 7:237–241
Fitzgerald RJ, Keyes PH (1960) Demonstration of the etiologic role of streptococci in experimental caries in the hamster. J Am Dent Assoc 61:9–19
Fukui K, Fukui Y, Moriyama T (1974) Purification and properties of dextransucrase and invertase from Streptococcus mutans. J Bacteriol 118:796–804
Fukui K, Kokeguchi S, Kato K, Miyaka Y, Nogami R, Moriyama T (1983) Immunochemical properties of glucosyltransferases from Streptococcus mutans. Infect Immun 39:762–766
Guggenheim B (1970) Extracellular polysaccharides and microbial plaque. Int Dent J 20:675–678
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 11:1252–1260
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 23:61–70
Hojo S, Higuchi M (1981) The role of insoluble glucan as a diffusion barrier in dental plaque. Jpn J Oral Biol 23:527–533
Keyes PH (1960) The infection and transmissible nature of experimental dental caries. Arch Oral Biol 1:304–320
Kuramitsu HK (1975) Characterization of extracellular glucosyltransferase activity of Streptococcus mutans. Infect Immun 12:738–749
Kuramitsu HK, Ingersoll L (1976) Immunological relationship between glucosyltransferases from Streptococcus mutans serotypes. Infect Immun 14:636–644
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 2:235–239
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Littleton NW, Kakehashi S, Fitzgerald RJ (1970) Recovery of specific “caries-inducing” streptococci from carious lesion in the teeth of children. Arch Oral Biol 15:461–463
Loesche WJ, Rowan J, Staffon LH, Loos PJ (1975) Association of Streptococcus mutans with human dental decay. Infect Immun 11:1252–1260
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 10:1135–1145
Oakley BR, Kirsch DR, Morris NR (1980) A simple ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem 105:361–363
Osborne RM, Lamberts BL, Meyer TS, Roush AH (1976) Acrylamide gel electrophoretic studies of extracellular sucrose-metabolizing enzymes of Streptococcus mutans. J Dent Res 55:77–84
Perch B, Kjems E, Ravn T (1974) Biochemical and serological properties of Streptococcus mutans from various human and animal sources. Acta Pathol Microbiol Scand 82:357–370
Russell RRB (1976) Classification of Streptococcus mutans strains by SDS gel electrophoresis. Microbios Lett 2:55–59
Russell RRB (1979) Use of Triton X-100 to overcome the inhibition of fructosyltransferase by SDS. Anal Biochem 97:173–175
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 201–210
Smith DJ, Taubman MA (1977) Antigenic relatedness of glucosyltransferase enzymes from Streptococcus mutans. Infect Immun 15:91–103
Suginaka H, Wada H, Kotani S (1975) Initial products in utilization of sucrose by Actinomyces viscosus. Biken J 18:271–274
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 95–97
Tanzer JM, Brown AT, MacInerney MF, Woodiel FN (1977) Comparative study of invertases of Streptococcus mutans. Infect Immun 16:318–327
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: 91–109
Tsumori H, Shimamura A, Mukasa H (1983) Comparative study of Streptococcus mutans extracellular glycosyltransferases by isoelectric focusing. J Gen Microbiol 129:3261–3269
Zacharius RM, Zell TE, Morrison JH, Wordlock JJ (1969) Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem 30:148–152
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kametaka, S., Hayashi, S., Miyake, Y. et al. Electrophoretic studies of extracellular glucosyltransferases and fructosyltransferases from seventeen strains of Streptococcus mutans . Arch. Microbiol. 147, 207–212 (1987). https://doi.org/10.1007/BF00463476
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00463476