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C L Wittenberger

Publications and source records attributed to C L Wittenberger.

11 recordsLinked to original sources
PubMed1979

Characterization of a phosphoenolpyruvate-dependent sucrose phosphotransferase system in Streptococcus mutans.

A phosphoenolpyruvate-dependent sucrose phosphotransferase system has been identified in Streptococcus mutans. Sucrose phosphotransferase activity was inducible by sucrose and had an apparent Km for sucrose of 70 microM. The product of the sucrose phosphotransferase reaction was isolated and identified as sucrose phosphate. Additional analysis revealed that the phosphate group was on the glucose moiety. Mutants unable to grow in media containing low concentrations of sucrose were isolated and found to be missing either sucrose phosphotransferase activity or the ability to hydrolyze sucrose phosphate.

Enzyme Induction↗
PubMed1979

Regulation and function of sucrose 6-phosphate hydrolase in Streptococcus mutans.

Sucrose catabolism by Streptococcus mutans is initiated by a phosphoenolpyruvate-dependent sucrose phosphotransferase reaction that produces sucrose 6-phosphate the latter is then cleaved by a sucrose 6-phosphate hydrolase reaction that yields glucose 6-phosphate and fructose. We have examined the regulation of the sucrose 6-phosphate hydrolase and found that it was synthesized constitutively whereas sucrose phosphotransferase activity was inducible. However, the levels of both sucrose phosphotransferase and sucrose 6-phosphate hydrolase were repressed when fructose was used as a growth substrate. The specific activity of sucrose 6-phosphate hydrolase in permeabilized cells was approximately 30 mmol/min per mg (dry weight of cells), and it had an apparent Km for sucrose 6-phosphate of 0.3 mM. analysis of a mutant that was missing sucrose 6-phosphate hydrolase activity revealed that its ability to hydrolyze sucrose was reduced.

Glucosephosphates↗
PubMed1978

Rapid method for identification and enumeration of oral Actinomyces.

Serotype-specific antisera prepared against whole cells of Actinomyces viscosus, A. naeslundii, and A. israeli were labeled with fluorescein dye and used to detect and quantitate antigenically related microorganisms in human dental plaque. By relating the DNA content of the dental plaque microflora to the number of Actinomyces present in the plaque samples, a reproducible method was developed for specifically enumerating five serotypic representatives of this genus found in human plaque.

Actinomyces↗
PubMed1978

Tween 80 effect on glucosyltransferase synthesis by Streptococcus salivarius.

Streptococcus salivarius (ATCC 25975) produced very low or nondetectable amounts of the extracellular enzyme glucosyltransferase (GTase) when grown in a chemically defined medium. The addition of Tween 80 to this medium resulted in the production of markedly enhanced levels of the enzyme. Oleic acid, the methyl ester of oleic acid, and sucrose each could not substitute for Tween 80 in this regard. The surfactant had no direct activating effect on performed enzyme activity. Tween 80 also stimulated the production of GTase by concentrated cells suspended in defined medium during a time when no measurable growth occurred. Under these conditions, the stimulatory effect of Tween 80 was blocked by chloramphenicol. It was further found that the surfactant dramatically stimulated the differential rate of GTase synthesis. These and other data strongly suggest that Tween 80 stimulates the production of extracellular GTase by acting either directly or indirectly at the level of enzyme synthesis.

Chloramphenicol↗
PubMed1977

Purification, resolution, and interaction of the glucosyltransferases of Streptococcus mutans 6715.

The extracellular glucosyltransferase produced by Streptococcus mutans 6715 was purified from culture supernatant fluids to a specific activity of 9.6 IU/mg of protein, with an overall recovery of 87%. The purified enzyme preparation, designated unfractionated enzyme (UFE), synthesized only water-insoluble glucans from sucrose during the initial stages of the reaction, although some water-soluble polymers accumulated after extended periods of incubation. It was free from measurable fructosyltransferase activity. The UFE preparation was resolved into two different catalytically active components by ethanol fractionation. One fraction (designated insoluble product enzyme [IPE]) synthesized water-insoluble glucans, whereas the other (designated soluble product enzyme [SPE]) produced primarily water-soluble glucans. The difference between the insoluble glucans made by the UFE preparation and those made by the IPE fraction appeared to be due to interaction of the SPE and IPE components in the UFE preparation. Addition of commercial dextrans or enzymatically prepared glucans to the glucosyltransferase assay altered the amounts of soluble and insoluble glucans synthesized by the UFE preparation. The molecular weights of the major enzymatically active proteins producing insoluble and soluble glucans were estimated by gel filtration chromatography to be 150,000 and 175,000, respectively.

Dextranase↗
PubMed1976

Multicomponent nature of the glucosyltransferase system of Streptococcus mutans.

The glucosyltransferases of S mutans 6715 were resolved into two major fractions. One fraction synthesized water-soluble glucans and the other made water-insoluble glucans. Each fraction was found by polyacrylamide gel electrophoresis to be composed of several catalytically active species, apparently glycoprotein in nature. Treatment of the glucosyltransferases with dextranase in the absence of sucrose caused an interconversion of enzyme forms concomitant with a time-dependent loss of enzyme activity, but did not appear to remove significant amounts of the carbohydrate associated with the enzymes. Comparison of enzyme activity patterns on polyacrylamide gels of the five different S mutans serotypes further emphasizes the complexity of the glucosyltransferase system from this group of microorganisms.

Chemical Phenomena↗
PubMed1975

Purification and properties of an NADP-specific 6-phosphogluconate dehydrogenase from Streptococcus faecalis.

A procedure is described for the purification of 6-phosphogluconate dehydrogenase (6-phospho-D-gluconate:NADP oxidoreductase (decarboxylating) EC 1.1.1.44) from cell extracts of Streptococcus gaecalis. A 180-fold purification was achieved with an over-all yield of about 12% and an average specific activity of 14. The enzyme was homogeneous as determined by polyacrylamide gel electrophoresis, immunoelectrophoresis, and sedimentation equilibrium, studies. Its weight average molecular weight, as measured by sedimentation equilibrium, was 108,000 +/- 3,600. Other methods employed for molecular weight determinations gave values that ranged between 106,000 and 115,000. An analysis of the enzyme by sodium dodecyl sulfate polyacrylamide gel electrophoresis showed it to be a dimer composed of subunits having equal molecular weight. The amino acid composition of the streptococcal enzyme is reported. The apparent Km values for NADP and 6-phosphogluconate were calculated from kinetic data and found to be 0.015 mM and 0.024 mM, respectively. Kinetic studies also indicated that the binding of one substrate did not affect the apparent affinity of the enzyme for the other substrate.

Amino Acids↗
PubMed1975

Mannitol transport in Streptococcus mutans.

A hexitol-inducible, phosphoenolpyruvate-dependent phosphotransferase system was demonstrated in Streptococcus mutans. Cell-free extracts obtained from mannitol-grown cells from a representative strain of each of the five S. mutans serotypes (AHT, BHT, C-67-1, 6715, and LM7) were capable of converting mannitol to mannitol-1-phosphate by a reaction which required phosphoenolpyruvate and Mg2+. Mannitol and sorbitol phosphotransferase activities were found in cell-free extracts prepared from cells grown on the respective substrate, but neither hexitol phosphotransferase activity was present in extracts obtained from cells grown on other substrates examined. A heat-stable, low-molecular-weight component was partially purified from glucose-grown cells and found to stimulate the mannitol phosphotransferase system. Divalent cations Mn2+ and Ca2+ partially replaced Mg2+, while Zn2+ was found to be highly inhibitory.

Alcohol Oxidoreductases↗