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At least 19 recordsLinked to original sources

Xylitol metabolism in xylitol-sensitive and xylitol-resistant strains of streptococci.

The metabolism of xylitol in xylitol-sensitive strains (strains whose growth is inhibited by xylitol) and xylitol-resistant strains (growth not inhibited) of oral streptococci was compared. Both xylitol-sensitive and xylitol-resistant strains took up xylitol. In the sensitive cells, the xylitol was probably transported via a phosphotransferase system. This resulted in intracellular accumulation of xylitol-5-phosphate and xylulose-5-phosphate. These metabolites were not detected in the xylitol-resistant strains, which probably transported xylitol via a permease system. It appeared that the resistant strains were able to utilize xylitol as carbon and energy source in the absence of other carbohydrates.

Drug Resistance, Microbial↗

Effects of xylitol, xylitol-sorbitol, and placebo chewing gums on the plaque of habitual xylitol consumers.

Xylitol reduces plaque but the reduction mechanism is largely unknown. The main aim of the present study was to determine whether the xylitol-induced reduction in the amount of plaque and the number of mutans streptococci could be demonstrated in subjects with (presumably) high levels of xylitol-resistant (XR; not inhibited by xylitol) mutans streptococci acquired following previous xylitol consumptions. 37 healthy dental students participated in the double-blind study. All subjects had been uncontrolled, habitual consumers of xylitol-containing products for at least 1 yr before the study. A 1-month washout period was followed by a 2-week test period during which either xylitol, xylitol-sorbitol or unsweetened chewing gum base was chewed 3-5 x a day. Plaque and saliva samples were collected at baseline and at the 2-week point for determination of the amount of plaque, microbiological variables, and hydrolytic enzymes. Mixtures of xylitol and sorbitol seemed to perform equally well with respect to reduction in the amount of plaque but not the number of mutans streptococci. Thus, polyols were the active ingredients of chewing gums able to modulate the amount of plaque and its microbial composition. Xylitol reduced plaque with a mechanism which appeared not to be associated with the study-induced changes in the proportion (%) of mutans streptococci in plaque, the number of salivary mutans streptococci, the proportion of XR strains in plaque or saliva, or the hydrolytic enzyme activities of plaque.

Adult↗

Xylitol concentrations in the saliva of children after chewing xylitol gum or consuming a xylitol mixture.

Xylitol prevents otitis media when given to children regularly five times per day. To find a more convenient dosing schedule, an enzymatic assay was used to measure xylitol concentrations in the saliva of 65 children after giving them xylitol chewing gum or syrup in doses equal to those used in clinical trials. Although concentrations high enough to have an antimicrobial effect were attained, the xylitol disappeared from the saliva within 15 min. This finding indicates that high peak concentrations are more important for efficacy than the amount of time the xylitol concentration exceeds that needed to produce an antimicrobial effect. A schedule with the same single doses given less frequently could be clinically effective in preventing otitis media.

Administration, Oral↗

Emergence of multiple xylitol-resistant (fructose PTS-) mutants from human isolates of mutans streptococci during growth on dietary sugars in the presence of xylitol.

The growth inhibition of mutans streptococci is one of the proposed mechanisms of action of xylitol, a caries-preventive natural carbohydrate sweetener. Xylitol is taken up and accumulated as non-metabolizable, toxic xylitol phosphate via a constitutive fructose PTS, and selects, during in vitro growth at the expense of glucose, for natural xylitol-resistant mutants that lack constitutive fructose PTS activity. Since long-term xylitol consumption leads to the emergence of xylitol-resistant mutans populations in humans in an oral environment containing sugars of dietary origin, we wanted to test the hypothesis that xylitol-resistant cells could be selected from mutans streptococci strains during in vitro growth on fructose, sucrose, or lactose. Three laboratory strains and three fresh mutans streptococcal isolates were repeatedly transferred in trypticase-yeast extract medium supplemented with glucose, fructose, sucrose, or lactose in the presence and absence of xylitol. Depending on the growth sugar, the presence of xylitol resulted in the selection of xylitol-resistant populations for several of the six strains tested, but not necessarily in the presence of all four sugars. All six strains rapidly became xylitol-resistant when grown on glucose in the presence of xylitol. All three fresh isolates became xylitol-resistant after 9 to 16 transfers in the presence of fructose or sucrose plus xylitol, while none of the laboratory strains became xylitol-resistant after 16 transfers in the presence of these sugars. The growth rates of 12 xylitol-resistant mutants in the presence of eight sugars suggested the existence of various types of xylitol-resistant mutants. The data partially explain the occurrence of xylitol-resistant mutans populations in long-term xylitol consumers and suggest a mechanism consistent with a selection process. Since various preliminary results suggest that xylitol-resistant natural mutants may be less virulent and less cariogenic than their parent strains, this selection process may alter, for the better, the mutans streptococci population of the plaque and play a role in the caries-preventive action of xylitol.

Culture Media↗

Intracellular xylitol-phosphate hydrolysis and efflux of xylitol in Streptococcus sobrinus.

The parental strain Streptococcus sobrinus (Streptococcus mutans ATCC 27352), which is known to transport, phosphorylate and accumulate xylitol intracellularly as nonmetabolizable xylitol-phosphate (xylitol-sensitive (XS) strain) and its xylitol-resistant (XR) spontaneous mutant were used to further investigate the inhibitory action of xylitol on oral streptococci. Fructose-grown XR cells did not accumulate xylitol-phosphate, indicating that the inducible fructose PTS is incapable of transporting the pentitol. The intracellularly accumulated pentitol-phosphate by the XS cells did not prevent the subsequent uptake and degradation of glucose or fructose, despite a drop in the PEP pool and a 50% inhibition of the glucose but not the fructose catabolism. Intracellular dephosphorylation of the pentitol-phosphate and release of xylitol in the extracellular medium resulted in a rapid decrease of the intracellular level of this nonmetabolizable product. A Mg(++)- or Mn(++)-independent sugar-phosphate hydrolysing activity capable of splitting xylitol-phosphate was demonstrated in both XS and XR strains. Preincubation in the presence of N1-ethylmaleimide (NEM) and xylitol or NEM and fructose resulted in the subsequent inhibition of both xylitol uptake and efflux. The efflux kinetic at various temperatures is compatible with a facilitated diffusion by the phosphotransferase system EIIfru without, however, excluding the existence of an additional exit route, but it excludes a simple diffusion exit process. The results are consistent with the existence of a xylitol futile cycle contributing to the growth inhibition of S. sobrinus by the pentitol without excluding a toxic effect of xylitol-phosphate. Discrepancies in the literature on the action of xylitol on S. mutans could be explained in the light of the evidence presented.

Dental Plaque↗

Xylitol concentration in saliva and dental plaque after use of various xylitol-containing products.

The study consisted of two sets of experiments, one in saliva and one in dental plaque. The xylitol concentration in saliva was determined enzymatically in 12 children (mean age 11.5 years) after a standardised use of various xylitol products: (A) chewing gums (1.3 g xylitol), (B) sucking tablets (0.8 g xylitol), (C) candy tablets (1.1 g xylitol), (D) toothpaste (0.1 g xylitol), (E) rinse (1.0 g xylitol), and (F) a non-xylitol paraffin. Unstimulated saliva was sampled 1, 3, 8, 16 and 30 min after use. The concentration in dental plaque was determined after mouthrinses with contrasting amounts of xylitol (LX = 2.0 g, HX = 6.0 g, and control) and supragingival plaque was collected and pooled after 5, 15 and 30 min. The mean xylitol concentration in saliva at baseline was approximately 0.1 mg/ml. All xylitol-containing products resulted in significantly increased levels (p < 0.05) immediately after intake and remained elevated for 8-16 min in the different groups. The highest mean value in saliva was obtained immediately after use of chewing gums (33.7 +/- 16.4 mg/ml) and the lowest was demonstrated after using toothpaste (8.2 +/- 4.9 mg/ml). No significant differences were demonstrated between chewing gums (A), sucking tablets (B), candy (C) and rinses (E). In dental plaque, the mean values were 8.6 +/- 5.4 and 5.1 +/- 4.0 mg/ml 5 min after HX and LX rinses. Concerning the higher concentration, the values remained significantly elevated (p < 0.05) during the entire 30-min follow-up. In conclusion, commonly advocated xylitol-containing products gave elevated concentrations of xylitol in unstimulated whole saliva and dental plaque for at least 8 min after intake.

Analysis of Variance↗

Selection for Streptococcus mutans with an altered xylitol transport capacity in chronic xylitol consumers.

The effect of long-term consumption of refined xylitol on the natural populations of S. mutans in the human oral cavity has been investigated. Fifty-four S. mutans strains were isolated from adults and children who had been consuming commercial food products containing xylitol for a period of from 1 1/2 to 10 years. Twenty isolates were also obtained from control subjects who had never consumed xylitol-containing commercial food products. The inhibitory effect of xylitol on the isolated strains was determined by monitoring growth on glucose in the presence or absence of xylitol. This was used to define the sensitivity of each isolate to xylitol. Phosphoenolpyruvate:sugar phosphotransferase (PEP-PTS) activities were measured by means of the soluble and membrane fractions prepared from strains from both study populations. It was found that 87% of the fresh isolates from xylitol consumers were xylitol-resistant (XR), compared with only 10% of the strains isolated from the control subjects. The XR strains had low constitutive fructose PTS activity and very low xylitol-phosphorylating capacity. The xylitol-sensitive (XS) strains, however, had much higher levels of constitutive fructose PTS activity and phosphorylated xylitol 16 times more rapidly than did the XR strains. Evidence for the phosphorylation of xylitol by a fructose PEP-PTS in the XS strains was obtained. The growth inhibition by the intracellular accumulation of non-metabolizable toxic xylitol phosphate and its prevention by the presence of fructose are discussed.

Adolescent↗

Development of a xylitol biosensor composed of xylitol dehydrogenase and diaphorase.

In preparation for the development of a xylitol biosensor, the xylitol dehydrogenase of Candida tropicalis IFO 0618 was partially purified and characterized. The optimal pH and temperature of the xylitol dehydrogenase were pH 8.0 and 50 degrees C, respectively. Of the various alcohols tested, xylitol was the most rapidly oxidized, with sorbitol and ribitol being reduced at 65% and 58% of the xylitol rate. The enzyme was completely inactive on arabitol, xylose, glucose, glycerol, and ethanol. The enzyme's xylitol oxidation favored the use of NAD+ (7.9 U/mg) over NADP+ (0.2 U/mg) as electron acceptor, while the reverse reaction, D-xylulose reduction, favored NADPH (7.7 U/mg) over NADH (0.2 U/mg) as electron donor. The K(m) values for xylitol and NAD+ were 49.8 mM and 38.2 microM, respectively. For the generation of the xylitol biosensor, the above xylitol dehydrogenase and a diaphorase were immobilized on bromocyan-activated sephallose. The gel was then attached on a dissolved oxygen electrode. In the presence of vitamin K3, NAD+ and phosphate buffer, the biosensor recorded a linear response to xylitol concentration up to 3 mM. The reaction was stable after 15 min. When the biosensor was applied to a flow injection system, optimal operation pH and temperature were 8.0 and 30 degrees C, respectively. The strengths and limitations of the xylitol biosensor are its high affinity for NAD+, slow reaction time, narrow linear range of detection, and moderate affinity for xylitol.

Biosensing Techniques↗

Uptake of 14C-xylitol by xylitol-cultured Streptococcus sobrinus ATCC 27352 and Streptococcus mitis ATCC 36249 in vitro.

The effect of three successive cultures in the presence of 6% xylitol on the uptake of 14C-xylitol was studied using resting cells of Streptococcus sobrinus ATCC 27352 and Streptococcus mitis ATCC 36249. In the case of S. mitis, the three successive cultures did not alter the growth inhibition observed in the presence of xylitol. In the case of S. sobrinus, however, growth inhibition decreased. The 14C-xylitol uptake experiments also demonstrated that uptake of xylitol by S. sobrinus was decreased by culture in the presence of xylitol. Previous 14C-xylitol uptake levels were, however, re-established by culturing S. sobrinus in the presence of glucose alone. Culture in the presence of xylitol did not affect 14C-xylitol uptake in the case of S. mitis. These results show that S. sobrinus and S. mitis differ in the ways they handle of exogenous xylitol, and that uptake of xylitol by S. sobrinus could be reversibly regulated by addition xylitol to the growth medium.

Cell Division↗

Uptake and expulsion of 14C-xylitol by xylitol-cultured Streptococcus mutans ATCC 25175 in vitro.

The effect of successive cultivations in the presence of 6% xylitol on the uptake and expulsion of 14C-xylitol was studied using the cells of Streptococcus mutans 25175. Three sequential cultivations did not alter the growth inhibition percentage (approximately 50%) observed in the presence of 6% xylitol. The 14C-xylitol uptake experiments performed with growing and resting cells showed that both the uptake and the expulsion of xylitol were enhanced by xylitol-culturing. Both xylitol-cultured and resting control cells contained only one major labeled compound which was identified as 14C-xylitol 5-phosphate. The label subsequently was expelled from the cells as 14C-xylitol. These results indicate that S. mutans possesses an intracellular xylitol cycle and this cycle is regulated by adding xylitol to the growth medium.

Carbon Radioisotopes↗

Effect of xylitol consumption on the plaque-saliva distribution of mutans streptococci and the occurrence and long-term survival of xylitol-resistant strains.

Since the exposure of mutans streptococci to xylitol is known to select for xylitol-resistant (XR) natural mutants, the occurrence and long-term survival of such xylitol-resistant strains was evaluated in a cross-sectional sampling of participants of the Ylivieska xylitol study four years after the original two-year experimental period. Paraffin-stimulated whole saliva was first collected, and then plaque was collected and pooled. The salivary and dental plaque mutans streptococci were enumerated after growth on TSY20B agar. The proportion of XR strains was determined by autoradiography with 14C-xylitol. A strong and significant correlation (r = 0.645 and p = 0.005) between the number of mutans streptococci in saliva and in dental plaque was observed in non-consumers of xylitol. Such a correlation totally disappeared (r = 0.098 and p = 0.612) in xylitol-exposed consumers (habitual and former xylitol-consumers). The proportion of the salivary XR mutants (35%) in non-consumers (n = 16) was significantly lower than in the xylitol-exposed consumers (79%) (n = 27), (p = 0.0001) or in former consumers (75%) (n = 13), (p = 0.0008) or in the habitual consumers (83%) (n = 14), (p = 0.004). The proportion of XR mutants in dental plaque was, on the average, much lower than in the corresponding saliva. The proportion of XR in the plaque of xylitol non-consumers was half of that of the xylitol-exposed group, but the difference was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

The effect of dietary xylitol on the ability of rat caecal flora to metabolise xylitol.

The effect of dietary xylitol on the ability of the rat caecal flora to metabolise xylitol was investigated. Xylitol metabolism in micro-organisms has generally been assessed in terms of pH change and acid production which are often insensitive in demonstrating low rates of substrate utilisation. Using a rapid and sensitive radioisotopic assay, in which 14CO2 production from [U-14C] xylitol was measured, it was possible to show that the caecal microflora obtained from rats can metabolise xylitol. This activity was increased 10, 15, 30 and 40-fold in the caecal flora taken from rats fed diets containing 2.5,5,10 and 20% xylitol, respectively. Using the caecal microflora of normally fed rats, the order of 14CO2 production from 14C-labelled sugars and sugar alcohols was glucose greater than fructose greater than xylose greater than sorbitol greater than or equal to xylitol. The feeding of glucose and fructose did not alter the 14CO2 producing activities, whereas xylose feeding increased xylose metabolism, sorbitol feeding increased sorbitol and xylitol metabolism, xylitol feeding increased sorbitol, xylose and its own metabolism and arabitol feeding increased xylose and sorbitol metabolism. Marked changes were also observed in the population of the caecal flora of xylitol-fed rats, with increases in the number of gram-positive bacteria, compared to rats on a normal diet. Possible mechanisms for these effects involve mutation, selection of micro-organisms capable of metabolising sugar alcohols, and the induction of enzymes involved in sugar alcohol metabolism.

Adaptation, Physiological↗

Cariogenic traits in xylitol-resistant and xylitol-sensitive mutans streptococci.

Long-term xylitol consumption leads to the emergence of xylitol-resistant (X-R) mutans streptococci. The aim of the present study was to compare cariogenic traits in X-R and xylitol-sensitive (X-S) strains. Six strains of mutans streptococci, three X-R and three X-S strains, were studied. Xylitol resistance and sensitivity were confirmed by growth in xylitol-supplemented media. Acid production from glucose or fructose or uptake of xylitol was initiated by adding (14)C-labelled glucose, fructose or xylitol to bacterial suspensions. The resultant metabolites were identified by HPLC. Lactate was the major metabolite from glucose, whether the bacteria were grown in the presence or the absence of xylitol. Lactate production per colony-forming unit was lower in X-S cells than in X-R cells. Fructose was metabolized by both X-R and X-S cells. Both X-R and X-S cells took up xylitol, but xylitol-5-P was detected in X-S cells only. Total polysaccharides were measured through production of C(14)-labelled ethanol-insoluble polymers from [U(14)-C]-sucrose. No difference in polysaccharide production was found between X-R and X-S cells. The present study thus does not support the contention that X-R are less cariogenic than X-S mutans streptococci.

Carbon Radioisotopes↗

Effect of xylitol-containing chewing gums on interdental plaque-pH in habitual xylitol consumers.

OBJECTIVE: The aim was to investigate the effect of high and low amounts of xylitol on the interdental plaque-pH, directly and after sucrose challenge, in schoolchildren with habitual consumption. MATERIAL AND METHODS: The study group consisted of 11 healthy children (10-15 years) with low caries risk and the experiment had a single-blind crossover (Latin square) design. After a 2-week run-in period with a daily 4.0 g xylitol intake, the children were subjected to single-dose exposures of chewing gums with (i) paraffin (CTR; no xylitol), (ii) low-dose xylitol (LX; 2.0 g xylitol), and (iii) high-dose xylitol (HX; 6.0 g xylitol) in a randomized order separated by a washout period of 1 week. Samples of chewing-stimulated whole saliva were collected prior to and after the experimental period for determination of bacterial counts. The outcome measures were in situ plaque-pH (micro-touch method) and area under the pH curve (AUC). RESULTS: The AUC was significantly greater (p < 0.05) in the HX group compared to the LX and control groups during the first 5 min after chewing. After a 10% sucrose rinse, the interdental plaque-pH dropped in all groups but the HX regimen displayed significantly less reduction 0-5 min after chewing (p < 0.05). No significant alterations of the total viable counts or mutans streptococci levels in saliva were disclosed during the 4-week experimental period. CONCLUSIONS: The present results suggested that a high single dose of xylitol had a short and limited beneficial effect on interdental plaque-pH in habitual xylitol consumers, while a low single dose, resembling normal chewing gum use, did not differ from the control.

Adolescent↗

A rare sugar xylitol. Part II: biotechnological production and future applications of xylitol.

Xylitol is the first rare sugar that has global markets. It has beneficial health properties and represents an alternative to current conventional sweeteners. Industrially, xylitol is produced by chemical hydrogenation of D-xylose into xylitol. The biotechnological method of producing xylitol by metabolically engineered yeasts, Saccharomyces cerevisiae or Candida, has been studied as an alternative to the chemical method. Due to the industrial scale of production, xylitol serves as an inexpensive starting material for the production of other rare sugars. The second part of this mini-review on xylitol will look more closely at the biotechnological production and future applications of the rare sugar, xylitol.

Biotechnology↗

Production of xylitol from D-xylose by a xylitol dehydrogenase gene-disrupted mutant of Candida tropicalis.

Xylitol dehydrogenase (XDH) is one of the key enzymes in d-xylose metabolism, catalyzing the oxidation of xylitol to d-xylulose. Two copies of the XYL2 gene encoding XDH in the diploid yeast Candida tropicalis were sequentially disrupted using the Ura-blasting method. The XYL2-disrupted mutant, BSXDH-3, did not grow on a minimal medium containing d-xylose as a sole carbon source. An enzyme assay experiment indicated that BSXDH-3 lost apparently all XDH activity. Xylitol production by BSXDH-3 was evaluated using a xylitol fermentation medium with glucose as a cosubstrate. As glucose was found to be an insufficient cosubstrate, various carbon sources were screened for efficient cofactor regeneration, and glycerol was found to be the best cosubstrate. BSXDH-3 produced xylitol with a volumetric productivity of 3.23 g liter(-1) h(-1), a specific productivity of 0.76 g g(-1) h(-1), and a xylitol yield of 98%. This is the first report of gene disruption of C. tropicalis for enhancing the efficiency of xylitol production.

Base Sequence↗

Selection and characterisation of a xylitol-derepressed Aspergillus niger mutant that is apparently impaired in xylitol transport.

Aspergillus niger is known for its biotechnological applications, such as the use of xylanase enzyme for the degradation of hemicellulose. Depending on culture conditions, several polyols may also be accumulated, such as xylitol during D: -xylose oxidation. Also during industrial fermentation of xylose for the production of fuel ethanol by recombinant yeast, xylitol is a by-product. We studied xylitol metabolism by isolating mutants that have impaired xylitol-mediated repression. Genetic and biochemical characterisation revealed that one of these mutants was affected not only in xylitol-mediated carbon repression, but also had impaired xylitol transport.

Alanine↗

Enhancement of xylitol productivity and yield using a xylitol dehydrogenase gene-disrupted mutant of Candida tropicalis under fully aerobic conditions.

The effects of glycerol and the oxygen transfer rate on the xylitol production rate by a xylitol dehydrogenase gene (XYL2)-disrupted mutant of Candida tropicalis were investigated. The mutant produced xylitol near the almost yield of 100% from D: -xylose using glycerol as a co-substrate for cell growth and NADPH regeneration: 50 g D: -xylose l(-1) was completely converted into xylitol when at least 20 g glycerol l(-1) was used as a co-substrate. The xylitol production rate increased with the O(2) transfer rate until saturation and it was not necessary to control the dissolved O(2) tension precisely. Under the optimum conditions, the volumetric productivity and xylitol yield were 3.2 g l(-1) h(-1) and 97% (w/w), respectively.

Aerobiosis↗