Impact of different fluoride concentrations and pH of dentifrices on tooth erosion/abrasion in vitro.

BACKGROUND: This in vitro study analysed the effect of different fluoride concentrations in acidic or neutral liquid dentifrices in protecting enamel and dentine from erosive and abrasive wear. METHODS: Bovine enamel and dentine specimens (n = 132) were randomly allocated to 11 groups (each n = 12): experimental liquid dentifrices with 550 ppm F, 1100 ppm F, 5000 ppm F or 0 ppm F/placebo (each at pH 4.5 and pH 7.0); and commercial dentifrices with 550 ppm F (Colgate Baby, pH 7.0), 1100 ppm F (Crest, pH 7.0) and 5000 ppm F (Duraphat, pH 7.0). The specimens were subjected to erosion for 90 seconds, 4 times/day, over 7 days. Immediately after the first and last erosion, the specimens were brushed for 15 seconds using one of the dentifrices. Tooth wear was measured profilometrically (µm) and analysed by ANOVA (p < 0.05). RESULTS: All fluoridated liquid dentifrices significantly reduced enamel wear compared to the placebo and commercial dentifrices. Only liquid dentifrices with 1100 and 5000 ppm F significantly reduced dentine wear compared to placebo dentifrice. The pH had no effect, but the consistency had a significant impact on the effect of dentifrices. CONCLUSIONS: Liquid dentifrices with high F concentration appear to be a good option to prevent tooth wear.

Biosynthesis of poly-beta-hydroxybutyrate and exopolysaccharides on Azotobacter chroococcum strain 6B utilizing simple and complex carbon sources.

Coproduction of poly-beta-hydroxybutyrate (PHB) and exopolysaccharides (EPS) was investigated with Azotobacter chroococcum strain 6B isolated from soil samples. The bacterium was cultured using various carbon sources solely or with 0.1 g/L of ammonium sulfate. Ammonium addition resulted in reduced PHB and EPS production with glucose, fructose, and sucrose media, but cellular mass remained constant except for sucrose. Protein was nearly twofold higher in ammonium-grown cultures. Glucose and fructose alone biosynthesized high amounts of EPS (maximum 2.1 and 1.1 g/L, respectively, at 72 h), whereas PHB was accumulated only in glucose-grown cells. Sucrose almost did not produce EPS. Conversely, PHB content was the highest obtained from all experimented conditions (1.1 g/L at 48 h, 40% cell dry wt). When a complex carbon source such as sugar cane molasses was utilized, PHB was accumulated concomitant with EPS production from the initial time to 48 h (0.75 g/L, 37% cell dry wt and 0.6 g/L, respectively), and then PHB decayed at 72 h (0.2 g/L). On the other hand, EPS continued to be biosynthesized (1.1 g/L, 72 h). PHB fractions of total intra- and extracellular biopolymers were calculated. Sucrose-modified Burk's medium without ammonium addition is suggested as a medium capable of diverting the carbon source for the production of intracellular PHB rather than EPS with A. chroococcum 6B.

[Isolation and characterization of Azotobacter sp. for the production of poly-beta-hydroxyalkanoates]. / Aislamiento y caracterización de Azotobacter sp. para la producción de poli-beta-hidroxialcanoatos.

A microorganism (Azotobacter sp.) was isolated from soil samples from the Agronomy Faculty campus and its ability to accumulate poly-beta-hydroxyalkanoate (PHAs) polymers was analyzed. The isolated strain (named No 8) accumulated 8 micrograms PHA/m of culture media as intracytoplasmic granules. The following properties of the strain were analyzed: utilization of different carbon sources, antibiotic resistance, optimal pH and temperature, pigment production, nitrogen fixation, proteolytic activity, acid and hydrogen sulphide production, optimal growth temperature, cyst formation, growth on phenol and sodium fluoride, catalase, pleomorfism and mobility. The synthesized polymer showed valuable properties respect to organic solvents resistance.

Aislamiento y caracterización de Azotobacter sp. para la producción de poli-beta-hidroxialcanoatos / [Isolation and characterization of Azotobacter sp. for the production of poly-beta-hydroxyalkanoates]

A microorganism (Azotobacter sp.) was isolated from soil samples from the Agronomy Faculty campus and its ability to accumulate poly-beta-hydroxyalkanoate (PHAs) polymers was analyzed. The isolated strain (named No 8) accumulated 8 micrograms PHA/m of culture media as intracytoplasmic granules. The following properties of the strain were analyzed: utilization of different carbon sources, antibiotic resistance, optimal pH and temperature, pigment production, nitrogen fixation, proteolytic activity, acid and hydrogen sulphide production, optimal growth temperature, cyst formation, growth on phenol and sodium fluoride, catalase, pleomorfism and mobility. The synthesized polymer showed valuable properties respect to organic solvents resistance.

Aislamiento y caracterización de Azotobacter sp. para la producción de poli-beta-hidroxialcanoatos. / [Isolation and characterization of Azotobacter sp. for the production of poly-beta-hydroxyalkanoates]

A microorganism (Azotobacter sp.) was isolated from soil samples from the Agronomy Faculty campus and its ability to accumulate poly-beta-hydroxyalkanoate (PHAs) polymers was analyzed. The isolated strain (named No 8) accumulated 8 micrograms PHA/m of culture media as intracytoplasmic granules. The following properties of the strain were analyzed: utilization of different carbon sources, antibiotic resistance, optimal pH and temperature, pigment production, nitrogen fixation, proteolytic activity, acid and hydrogen sulphide production, optimal growth temperature, cyst formation, growth on phenol and sodium fluoride, catalase, pleomorfism and mobility. The synthesized polymer showed valuable properties respect to organic solvents resistance.

L-serine production by a methylotroph and its related enzymes.

The production process of L-serine from methanol and glycine has been developed using a methylotroph with the serine pathway. Consecutive reactions of two enzymes, methanol dehydrogenase (MDH) and serine hydroxymethyltransferase (SHMT) are involved in the production. We screened a high producer, Hyphomicrobium methylovorum, which is an obligate methylotroph. With resting cells of the bacterium, 24 mg/ml of L-serine was produced from 100 mg/ml of glycine and 48 mg/ml of methanol in 3 days under optimal conditions. Next, a glycine-resistant mutant GM2 showed improved serine production (32-34 mg/ml). The mutant GM2 was found to have elevated activities of MDH and SHMT. Since there has so far been little report on the systematic characterization of enzymes of the serine pathway in methylotrophs, not only the above two enzymes but also the other three enzymes in H. methylovorum were purified and characterized: MDH, SHMT and hydroxypyruvate reductase (HPR) were crystallized; serine-glyoxylate aminotransferase (SGAT) and glycerate kinase (GK) were purified to homogeneity. As a result, all these enzymes were found to be stable against preservation and to exist abundantly in the bacterium. The gene of SHMT was cloned and its deduced amino acid sequence had homology to those of Escherichia coli (55%) and rabbit liver (44%), whereas the enzyme of the bacterium was immunochemically distinguishable from those of microorganisms other than Hyphomicrobium strains and mammalian livers.

Purification and characterization of hydroxypyruvate reductase from a serine-producing methylotroph, Hyphomicrobium methylovorum GM2.

Hydroxypyruvate reductase of a serine-producing methylotroph, Hyphomicrobium methylovorum GM2, was purified to complete homogeneity, crystallized and characterized, the first time for an enzyme from a methylotroph. The enzyme was found to be a dimer composed of identical subunits (38 kDa), the molecular mass of the enzyme being about 70 kDa. The enzyme was stable against heating at 25 degrees C for 10 min at pH values between 5 and 9. Optimal activity was observed at pH 6.8 and around 45 degrees C. The enzyme catalyzed the reduction of hydroxypyruvate with the oxidation of only NADH. Other than hydroxypyruvate, only glyoxylate served as a substrate. The Km values were found to be 0.175 mM for hydroxypyruvate and 10.8 mM for glyoxylate. Taking advantage of the high substrate specificity of this enzyme, a means of enzymatic determination of hydroxypyruvate was established.

Purification and characterization of a serine hydroxymethyltransferase from an obligate methylotroph, Hyphomicrobium methylovorum GM2.

A serine hydroxymethyltransferase was purified to complete homogeneity from a serine-producing obligate methylotroph, Hyphomicrobium methylovorum GM2. The enzyme has a molecular mass of about 98 kDa and consists of two subunits of identical molecular mass. The holoenzyme exhibits absorption maxima at 280 nm, 340 nm and 415 nm in potassium phosphate buffer, pH 7.3, the last of which shifts with a change in pH (6.0-7.5) and contains 2 mol pyridoxal phosphate/mol enzyme. The holoenzyme is converted to the apoenzyme on incubation with phenylhydrazine and reconstituted on the addition of pyridoxal phosphate. The enzyme activity was inhibited on the addition of several sulfhydryl-modifying reagents and then recovered with 2-mercaptoethanol. One sulfhydryl group per subunit was found to be responsible for the activity. Isoelectric focusing showed that the enzyme has a pI of 5.6. The Km values for glycine, L-serine and DL-beta-phenylserine are 0.046 mM, 0.15 mM and 33 mM respectively.

Crystalline serine hydroxymethyltransferase from an obligate methylotroph, Hyphomicrobium methylovorum.

Optimal culture conditions of a methylotrophic Hyphomicrobium methylovorum and improved purification of serine hydroxymethyltransferase from the bacterium were established for the large-scale preparation of the enzyme. The first crystalline serine hydroxymethyltransferase from the microbial source was obtained in the apo form and found to be homogeneous. Amino acid analysis revealed that the enzyme had higher value per subunit for acidic and neutral amino acids than that from rabbit liver. The carboxy-terminal amino acid analysis suggested the sequence -Ile-Ala-Tyr.