Adherence of
            <i>Veillonella</i>
            Species Mediated by Extracellular Glucosyltransferase from
            <i>Streptococcus salivarius</i>

McCabe, R.M.; Donkersloot, Jacob A.

doi:10.1128/iai.18.3.726-734.1977

Cited by 45 publications

(19 citation statements)

References 44 publications

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“…A significant correlation was found between the population of S. salivarius and the water-insoluble ^'^G-G'-polysaccharides. It is therefore possible that GTF's from this strain may be of clinical importance, as has been proposed previously (11,21). Subjects highly infected with S. mutans appeared to possess particularly high salivary transferase activity.…”

Section: Discussionsupporting

confidence: 63%

Polysaccharide production by cell free transferases in saliva in relation to salivary microflora

Scheie

Rölla

1984

European J Oral Sciences

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Scheie AA, Rolla G: Polysaccharide production by cell free transferases in saliva in relation to salivary microflora. Scand J Dent Res 1984; 92: 43-9. Abstract -The supernatant of centrifuged whole saliva was incubated with radiolabeled sucrose to measure polysaccharide production by cell free transferases and to examine whether waterinsoluble polysaccharides were produced. Amounts of polysaccharides were considered to reflect the level of cell free transferases in saliva. Plating samples on blood agar, MS and MSB plates gave salivary counts of total CFU, total streptococci, S. salivarius and S. mutans. The capability ofthe cell free portion of saliva to produce polymers was confirmed and it appeared that the cell free transferases were able to produce water-insoluble polysaccharides. Significant correlations were found between the total and the insoluble polysaccharides from '*C-G'-sucrose and total CFU, total streptococci, S. salivarius and S. mutans, respectively. Heavily S. mutans infected subjects seemed to produce particularly large amounts of water-insoluble polysaccharides from '*C-G'sucrose. The apparently water-insoluble '*C-F-polysaccharides correlated significantly to the number of S. salivarius. It was thus concluded that the constitution of the oral microflora and particularly the levels of S. mutans and S. salivarius were of importance for the level of cell free transferase activity.

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Section: Discussionsupporting

confidence: 63%

Polysaccharide production by cell free transferases in saliva in relation to salivary microflora

Scheie

Rölla

1984

European J Oral Sciences

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“…Streptococcus mutans produces at least three Gtfs: GtfB, which synthesizes mostly insoluble glucan (a1,3-linked); GtfC, which synthesizes a mixture of insoluble and soluble glucan (a1,6-linked); and GtfD, which synthesizes soluble glucan (as reviewed in Loesche 1986). The glucosyltransferases (Gtfs) secreted by this bacterium bind avidly to the pellicle formed on the tooth surface and to bacterial surfaces in an active form (McCabe and Donkersloot 1977;Schilling and Bowen 1988;Vacca-Smith and Bowen 1998;Hannig et al 2008). The glucans synthesized in situ promote the binding and accumulation of microorganisms on the apatitic surface and to each other (Schilling and Bowen 1992;Cross et al 2007) and are essential for the formation of cariogenic biofilms (Tanzer et al 1985;Yamashita et al 1993;Marsh 2003).…”

Section: Introductionmentioning

confidence: 99%

Structural organization and dynamics of exopolysaccharide matrix and microcolonies formation byStreptococcus mutansin biofilms

Xiao

Koo

2009

Journal of Applied Microbiology

101

126

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Aims: To investigate the structural organization and dynamics of exopolysaccharides (EPS) matrix and microcolonies formation by Streptococcus mutans during the biofilm development process. Methods and Results: Biofilms of Strep. mutans were formed on saliva‐coated hydroxyapatite (sHA) discs in the presence of glucose or sucrose (alone or mixed with starch). At specific time points, biofilms were subjected to confocal fluorescence imaging and computational analysis. EPS matrix was steadily formed on sHA surface in the presence of sucrose during the first 8 h followed by a threefold biomass increase between 8 and 30 h of biofilm development. The initial formation and further development of three‐dimensional microcolony structure occurred concomitantly with EPS matrix synthesis. Tridimensional renderings showed EPS closely associated with microcolonies throughout the biofilm development process forming four distinct domains (i) between sHA surface and microcolonies, (ii) within, (iii) covering and (iv) filling the spaces between microcolonies. The combination of starch and sucrose resulted in rapid formation of elevated amounts of EPS matrix and faster assembly of microcolonies by Strep. mutans, which altered their structural organization and susceptibility of the biofilm to acid killing (vs sucrose‐grown biofilms; P < 0·05). Conclusions: Our data indicate that EPS modulate the development, sequence of assembly and spatial distribution of microcolonies by Strep. mutans. Significance and Impact of the Study: Simultaneous visualization and analysis of EPS matrix and microcolonies provide a more precise examination of the structural organization of biofilms than labelling bacteria alone, which could be a useful approach to elucidate the exact mechanisms by which Strep. mutans influences oral biofilm formation and possibly identify novel targets for effective antibiofilm therapies.

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“…Cultures were grown overnight in Jordan broth with 5 mM glucose (11) or in the chemically defined FMC medium of Terleckyj et al (38) with 0.2% glucose, 0.2% Na2CO3, and 0.05% (vol/vol) Tween 80 (39,40). After centrifugation at 15,000 x g, supernatant fluids were adjusted to pH 6.5 and assayed for insolubleand soluble-glucan synthesis and fructosyltransferase (FT) with specifically labeled sucrose as described previously (28). Because the linkages in the polymers synthesized by these enzymes have not been characterized, we prefer not to identify the LM-7 GT and FT activities as dextransucrase (EC 2.4.1.5) and levansucrase (EC 2.4.1.10), respectively (40).…”

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confidence: 99%

“…The amount of product formed was proportional to the amount of supernatant fluid added. Assay results were converted to international units (28). For cell-associated GT assays, cells were washed twice with 0.9% NaCl, resuspended in 0.05 M potassium phosphate buffer (pH 6.5) at an apparent absorbance (600 nm) of 1.0, and assayed for insoluble-glucan synthesis (28) in the presence of 10 mM NaF.…”

mentioning

confidence: 99%

“…Assay results were converted to international units (28). For cell-associated GT assays, cells were washed twice with 0.9% NaCl, resuspended in 0.05 M potassium phosphate buffer (pH 6.5) at an apparent absorbance (600 nm) of 1.0, and assayed for insoluble-glucan synthesis (28) in the presence of 10 mM NaF. To study sucrose-induced agglutination of the washed cells, sucrose and sodium azide were added to final concentrations of 1 and 0.04%, respectively, and the suspensions were inspected visually for agglutination during incubation at 350C.…”

mentioning

confidence: 99%

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Characterization of glucosyltransferase-deficient, plasmid-containing mutants of Streptococcus mutans LM-7

Donkersloot¹,

Flatow²,

Gibson³

et al. 1978

Infect Immun

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The possibility that glucosyltransferase (GT)-mediated insoluble-glucan synthesis from sucrose is controlled by the 3-megadalton plasmid pAM7 in Streptococcus mutans LM-7 has been examined. A low-sucrose agar medium was developed to readily detect and quantitate presumptive GT-negative mutants. Such mutants were isolated from Todd-Hewitt broth cultures grown either with or without sodium dodecyl sulfate (10 [Lg/ml) or acriflavine (0.5 ig/ml) at frequencies ranging from about 0.01 to 1%. Independently isolated mutants had the following characteristics: (i) cells were virtually devoid of cell-associated GT and did not aggregate upon addition of sucrose; (ii) cell-free culture fluids synthesized 1OX less insoluble glucan than those of the parent; and (iii) cultures grown with sucrose did not form adherent deposits on the wall of the culture tube, as is typical of S. mutans. Both parent and mutants formed relatively little soluble glucan in 1-h assays. Three independently isolated mutants and the parent were found to contain similar amounts of plasmid DNA. Analysis by sucrose density gradient centrifugation and agarose gel electrophoresis did not reveal a size difference between the plasmids from parent and mutants. These results show that (i) S. mutans LM-7 generates GT-deficient mutants at relatively high frequency that still contain a 3-megadalton plasmid; (ii) both cell-associated and extracellular GT levels are depressed in the mutants, which suggests that these activities are directly or indirectly controlled by the same gene or by genes that segregate as a unit.About 10 years ago, several investigators demonstrated that the smooth-surface cariogenicity of Streptococcus mutans in animal model systems is associated with the formation of an adherent and water-insoluble glucan containing primarily a-1-6-and a-1, 3-linkages (16, 19). This concept has since been amply confirmed. The glucosyltransferase (GT) activity responsible for insoluble-glucan synthesis from sucrose has been relatively refractile to biochemical and genetic characterization. This appears to be at least partly due to its tendency to form complexes with itself and other sucrose-and dextran-metabolizing enzymes with apparent molecular weights greater than 106 (3,14,26,31). However, at least three major and several minor forms of the enzyme have been isolated from strain OMZ-176 (18), and electrophoretically distinct activities with molecular weights ranging from about 160,000 to 225,000 have since been reported from the genetically and biochemically related strain 6715 (3). It is not known whether the observed enzyme multiplicity is due to multiple GT genes or to posttranscriptional modification of a single GT gene product. The location of the gene(s) controlling insoluble-glucan synthesis is also open to question. Whereas it was first proposed that this trait is associated with an inducible prophage (17, 24), later evidence suggested that GT synthesis is controlled by a plasmid because plasmid-curing agents induced mutants at high frequency that s...

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Adherence of Veillonella Species Mediated by Extracellular Glucosyltransferase from Streptococcus salivarius

Cited by 45 publications

References 44 publications

Polysaccharide production by cell free transferases in saliva in relation to salivary microflora

Polysaccharide production by cell free transferases in saliva in relation to salivary microflora

Structural organization and dynamics of exopolysaccharide matrix and microcolonies formation byStreptococcus mutansin biofilms

Characterization of glucosyltransferase-deficient, plasmid-containing mutants of Streptococcus mutans LM-7

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