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Preparation of glucosyl transferase ec 2.4.1.5 from streptococcus mutans by evolution from water insoluble poly saccharide with a dissociating solvent


, : Preparation of glucosyl transferase ec 2.4.1.5 from streptococcus mutans by evolution from water insoluble poly saccharide with a dissociating solvent. Infection and Immunity 23(2): 446-452

Glucosyltransferase [GTF] (EC 2.4.1.5) was obtained by dissociation from water-insoluble polysaccharide in the presence of 6 M guanidine-hydrochloride. Water-insoluble polysaccharide was synthesized by cell-free culture supernatants from S. mutans strain 6715. Gel filtration of the GTF on a column of 8% agarose in phosphate buffer, followed by filtration on a column of 4% cross-linked agarose in 6 M guanidine-hydrochloride, gave a 23-fold enrichment of the enzyme. The enriched GTF preparation contained 22% carbohydrate and eluted at a position corresponding to a MW of 422,000. Polyacrylamide gel (5%) electrophoresis of this preparation revealed 2 regions which stained for protein, formed water-insoluble polysaccharide in the presence of sucrose, and precipitated with [hamster] antisera directed to crude GTF preparations. The guanidine-eluted enzyme could be primed by 5 .times. 10-5 M dextran T10 (MW 10,000). High-MW glucan, a possible glucan-binding protein and GTF were obtained after the final gel filtration step (4% cross-linked agarose). [The ability to obtain GTF free of other antigens (Ag) in a yield sufficient for immunization experiments may help resolve the role of GTF as a protective Ag in vaccines.].

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Related references

Smith, D.J.; Taubman, M.A.; Ebersole, J.L., 1979: Preparation of glucosyltransferase from Streptococcus mutans by elution from water-insoluble polysaccharide with a dissociating solvent. Glucosyltransferase (EC 2.4.1.5) was obtained by dissociation from water-insoluble polysaccharide in the presence of 6 M guanidine-hydrochloride. Water-insoluble polysaccharide was synthesized by cell-free culture supernatants from Streptococcus m...

Ciardi J.E.; Wittenberger C.L., 1974: Glucosyl transferase and poly saccharide production by streptococcus mutans. Federation Proceedings 33(5 PART 2): 1435

Evans R.T.; Emmings F.G.; Genco R.J., 1975: Preparation characterization and immunogenicity of insoluble poly saccharide antigen from cariogenic streptococcus mutans. Abstracts of the Annual Meeting of the American Society for Microbiology 75: 62

Figures, W.R.; Edwards, J.R., 1981: D glucosyl transferase ec 2.4.1.5 of streptococcus mutans isolation of 2 forms of the enzyme that bind to insoluble dextran. The D-glucosyltransferase from S. mutans 6715 was separated into 3 enzymic fractions that differ in their binding to dextran and in their synthesis of dextran from sucrose. One enzymic fraction (AFF-I) does not bind to insoluble dextran and it pro...

Smith E.E.; Ono K., 1981: Oligo saccharide precursors in the de novo synthesis of alpha glucan by a glucosyl transferase of streptococcus mutans. Federation Proceedings 40(6): 1597

Sato, S.; Koga, T.; Yakushiji, T.; Nagasawa, S.; Inoue, M., 1982: Effects of exogenous soluble dextrans on insoluble glucan synthesis by streptococcus mutans glucosyl transferase ec 2.4.1.5. Production of water-insoluble glucan (ISG) from sucrose by cell-free S. mutans strain AHT glucosyltransferase (GTF; EC 2.4.1.5), first rapidly increased, and then sharply declined, as the amounts of water-soluble Dextrans T20-T500 present, were in...

Anonymous, 1982: Synthesis of adherent insoluble glucan by the concerted action of the 2 glucosyl transferase ec 2415 components of streptococcus mutans

Anonymous, 1979: Purification of streptococcus mutans glucosyl transferase ec 2415 by poly ethylene glycol precipitation

Fukushima K.; Motoda R.; Takada K.; Ikeda T., 1981: Resolution of streptococcus mutans glucosyl transferases into 2 components essential to water insoluble glucan synthesis. Febs Letters 128(2): 213-216

Kuramitsu, H.K.; Ingersoll, L., 1978: Antigenic relationships between the glucosyl transferase activities of streptococcus mutans ssp mutans. Antibodies, directed against the partially-purified glucosyltransferase-A (synthesizing primarily insoluble glucans) and purified glucosyltransferase-B (producing predominantly soluble glucans) of S. mutans GS5 (serotype c), were used to assess th...