Purification and properties of dextransucrase from Streptococcus sanguis

Carlsson, Jan; Newbrun, Ernest; Krasse, Bo

doi:10.1016/0003-9969(69)90140-x

Cited by 126 publications

(71 citation statements)

References 19 publications

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“…3). Our results concerning solution phase GtfSs are in agreement with those determined by Carlsson et al [1969], who found the temperature optimum for this enzyme in solution to be 45°C.…”

Section: Effect Of Temperature On the Activity Of Gtfss In Solution Asupporting

confidence: 82%

“…In previous studies by Carlsson et al [1969], the pH optimum for this enzyme in solution was found to be between 5.2 and 7.0. We measured the pH optimum of GtfSs in solution and on the surface of sHA beads.…”

Section: Effect Of Ph On the Activity Of Gtfss In Solution And Adsorbmentioning

confidence: 99%

“…Much of the recent information available concerning the role of glucan in the formation of experimental dental plaque biofilms has been obtained using Gtfs from Streptococcus mutans and Streptococcus sobrinus, and scant attention has been paid to the possible role of Gtf from Streptococcus sanguis (GtfSs). However, earlier studies were performed in which the GtfSs was purified and characterized in solution [Carlsson et al, 1969;Newbrun, 1971;Sharma et al, 1974;Beeley and Black, 1977]. These studies revealed that in solution, the GtfSs had a pH optimum between 5.2 and 7.0, a K m of 4.8 mmol/l sucrose, optimal activity at 45°C, and an isoelectric point at pH 7.9, and produced glucan with both α-1,6 and α-1,3 linkages.…”

Section: Saturation Of Sha Beads By Gtfssmentioning

confidence: 99%

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Studies Concerning the Glucosyltransferase of <i>Streptococcus sanguis</i>

et al. 2000

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We have shown in previous studies that the glucosyltransferase (Gtf) enzymes of Streptococcus mutans have distinct properties when adsorbed to a surface. In the present study, we compared the activity of Gtf from Streptococcus sanguis, designated GtfSs, in solution and on the surface of saliva–coated hydroxyapatite (sHA) beads, and determined the ability of its product glucan to support the adherence of oral microorganisms. Gtf from S. sanguis 804 NCTC 10904 was purified from culture supernatant fluids by means of hydroxyapatite chromatography. Enzyme and the substrate were prepared in buffers at pH values from 3.5 to 7.5. Maximum activity of GtfSs occurred between pH 5.5 and pH 6.5, whether in solution or adsorbed onto a surface. The solubilized and insolubilized enzymes showed highest activity at 40°C; activity was reduced by 50(±2)% at 20 and 30°C. The enzyme did not form glucans in either phase at 10 or 60°C. The K_m, determined from Lineweaver–Burk plots, for the enzyme in solution was 4.3(±0.4) mmol/l sucrose, and the K_m for the enzyme on sHA beads was 5.0(±1.0) mmol/l sucrose. The ability of the GtfSs glucan synthesized on the surface of sHA beads to support the adherence of oral bacteria was investigated. ³H–thymidine–labeled bacteria (S. mutans GS–5, S. sobrinus 6715, S. sobrinus 6716, S. sanguis 10904, Actinomyces viscosus OMZ105E, A. viscosus 2085, and A. viscosus 2086) were incubated with sHA beads coated with GtfSs glucan. S. mutans GS–5 displayed the highest level of binding numerically. These results show that the GtfSs of S. sanguis is active on sHA beads, that the pH optimum for activity on a surface differs slightly from that in solution, and that its product glucan can support the adherence of oral microorganisms.

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“…3). Our results concerning solution phase GtfSs are in agreement with those determined by Carlsson et al [1969], who found the temperature optimum for this enzyme in solution to be 45°C.…”

Section: Effect Of Temperature On the Activity Of Gtfss In Solution Asupporting

confidence: 82%

Section: Effect Of Ph On the Activity Of Gtfss In Solution And Adsorbmentioning

confidence: 99%

Section: Saturation Of Sha Beads By Gtfssmentioning

confidence: 99%

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Studies Concerning the Glucosyltransferase of <i>Streptococcus sanguis</i>

et al. 2000

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“…Indigenous strains of Streptococcus mutans serotype c isolated from the volunteers were grown in dialysate medium (14), and 100-mg samples of lyophilized killed bacteria were packed into gelatin capsules (11). These were enterically coated with cellulose acetate phthalate (9 g) dissolved in 90 ml of acetone/2 ml of triethyl citrate/3 ml of distilled water.…”

Section: Methodsmentioning

confidence: 99%

IgA antibody-producing cells in peripheral blood after antigen ingestion: evidence for a common mucosal immune system in humans.

Czerkinsky¹,

Prince²,

Michalek³

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

233

115

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The finding that ingestion of antigens results in the selective induction of IgA antibodies in external secretions suggests that antigen sensitizes Peyer's patch lymphoid cells, which migrate to mucosal sites and generate local secretory IgA (S-IgA) antibody responses. Evidence for a common mucosal immune system in humans has been scanty because of the difficulty in demonstrating migratory behavior of Peyer's patch cells. In the present study, peripheral blood mononuclear cells (PBMC) from human volunteers who had ingested capsules containing killed Streptococcus mutans were assayed for spontaneous antibody-producing cells. Four of five volunteers exhibited circulating IgA-producing cells within 7 days and reached maximum responses by days 10-12. One

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“…The procedure for serotyping and DNA fingerprinting using restriction endonuclease analysis was as described before ]: (1) Colonies representing all morphological types were picked and transferred to separate tubes of dialysed tryptose yeast medium [Carlsson et al, 1969]. (2) From this medium, one portion was taken for Gram's stain control.…”

Section: Bacterial Analysismentioning

confidence: 99%

Genotypes of Mutans Streptococci Tend to Persist in Their Host for Several Years

Thornqvist

2000

Caries Res

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The aim of this study was to evaluate the consistency of the prevalence of mutans streptococci in a group of Swedish families. Eleven families, which had previously been examined for genotypes of mutans streptococci, were re–examined after 2–5 years. The families consisted of mother, father and a child (mean age 7.2 years at the follow–up examination). One father did not participate. Pooled plaque samples were obtained from buccal and occlusal surfaces. Isolates of mutans streptococci were genotyped using chromosomal DNA digested with restriction endonuclease HaeIII, separated by gel electrophoresis and visualised through UV illumination after ethidium bromid staining. Comparing the DNA fingerprints of mutans streptococci found at baseline and follow–up, 9 children harboured one or two genotypes which were similar on the two sampling occasions. Two of these children had also gained a genotype. The remaining 2 children had lost a genotype each and 1 of them had gained two new genotyes. All 21 adults showed one or two genotypes identical to those found at baseline. Nine of these 21 adults had also lost one genotype. Four of these 9 and additionally 4 of the remaining adults showed one or two new genotypes. Six mother–child pairs shared a genotype at baseline and this pattern remained for five pairs at the end of the study. The results suggest that genotypes of mutans streptococci have a fairly high degree of consistency in children between 3 and 8 years of age as well as in adults, indicating persistence of the strains. However, the results also indicate that some subjects may gain and/or lose genotypes.

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Purification and properties of dextransucrase from Streptococcus sanguis

Cited by 126 publications

References 19 publications

Studies Concerning the Glucosyltransferase of <i>Streptococcus sanguis</i>

Studies Concerning the Glucosyltransferase of <i>Streptococcus sanguis</i>

IgA antibody-producing cells in peripheral blood after antigen ingestion: evidence for a common mucosal immune system in humans.

Genotypes of Mutans Streptococci Tend to Persist in Their Host for Several Years

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