Glucosyltransferase Enzyme Activity Research Articles

Inhibitory effect of nano Stolephorus insularis and calcium hydroxide on glucosyltransferase (GTF) activity of Lactobacillus aciophilus

Background: Dental caries is the most common multifactorial infectious disease worldwide which referes to the process of destruction tooth hard tissue caused by bacterial by-products in the form of acids through carbohydrate fermentation. Lactobacillus acidophilus is one of the main cariogenic bacteria that causes caries by forming biofilms with the help ofglucosyltransferase (GTF) enzymes. Calcium hydroxide has limited antibacterial effects. Nano brown anchovy contains fluor as the active compound which has the potential as an antibacterial alternative agent. Purpose: To explain the inhibition of the combination of nano Stolephorus insularis and calcium hydroxide on the activity of the glucosyltransferase enzyme of Lactobacillus acidophilus bacteria. Methods: This study used a combination of nano Stolephorus insularis 3.125% with calcium hydroxide as a treatment and aquades as a control. The GTF enzyme was obtained from the supernatant centrifuged by Lactobacillus acidophilus in BHIB. The activity of the GTF enzyme was considered by calculating the fructose levels using High Performance Liquid Chromatography (HPLC) in a certain formula.Results: Decreased levels of fructose was obtained in the treatment group. From the results of Mann- Whitney data analysis, there were significant difference in the study groups. Conclusion: the combination of nano Stolephorus insularis with calcium hydroxide is effective to inhibit the glucosyltransferase enzyme activity of Lactobacillus acidophilus bacteria.

An effective concentration of propolis extract to inhibit the activity of Streptococcus mutans glucosyltransferase enzyme

Background: According to Riset Kesehatan Dasar (Riskesdas) (2013) and the World Health Organisation (WHO), caries is still a global problem and highly prevalent in Indonesia. Caries is mainly caused by Streptococcus mutans with virulence factors known as glucosyltransferase (GTF). The GTF enzyme contribute to the pathogenesis of caries by converting sucrose to fructose and glucan, which are then used in the formation of biofilms and dental plaques. Natural propolis compounds containing flavonoids, terpenoids, saponins and tannins, can inhibit GTF enzyme activity. Purpose: This study aimed to determine an effective concentration of propolis extract for inhibiting the S. mutans GTF enzyme activity. Methods: This study used propolis extract at 14 μg/ mL, 16 μg/m and 1 μg/mL to determine the inhibitory effect on S. mutans GTF enzyme activity. The GTF enzyme were obtained from the supernatant from S. mutans culture centrifugation. The GTF enzyme activity was measured using high-performance liquid chromatography (HPLC) to calculate the fructose level. Results: The mean fructose concentration at 14 μg/mL, 16 μg/mL, and 18μg/mL were 3.31%, 1.56%, and 0.29%, respectively. Conclusion: The most effective concentration of propolis extract for inhibiting the effect of S. mutans GTF enzyme activity is 14 μg/mL.

EFFECT OF KARAMUNTING LEAF EXTRACT (Melastoma malabathricum L.) ON GLUCOSYLTRANSFERASE ENZYME OF Streptococcus mutans

ABSTRACTBackground: Karamunting leaf extract has secondary metabolite compounds such as tannin, flavonoid, and phenol that have antibacteria potential to be used as an alternative to mouthwash in addition to Chlorhexidine 0.12% to lower the incidence of tooth caries. Caries is caused by several important virulence factors, including Streptococcus mutans Glucosyltransferase (GTF) enzyme. Karamunting leaf extract can inhibit the work of GTF enzyme by precipitating, denaturing protein and damaging cell walls. Inhibition of Streptococcus bacteria enzyme activity will decrease plaque formation, thus decreasing the potential of dental caries. Objective: The purpose of this study was to determine the effect of Karamunting leaf extract and 0,12% Chlorhexidine on Glucosyltransferase enzyme activity of Streptococcus mutans. Methods: This study used true experimental design with post test only with control group design using 7 groups, including Karamunting leaf extract group with 5%, 10%, 15%, 20%, and 25% concentrations to determine the activity of S. mutans GTF enzyme. Results: The results showed that Karamunting leaf extract with 20% concentration was able to decrease the activity of Glucosyltransferase enzyme in Streptococcus mutans. Conclusion: Karamunting leaf extract affects Streptococcus mutans Glucosyltransferase enzyme activity. Keywords: Chlorhexidine 0,12%, Glucosiltransferase, Melastoma malabathricum L., Streptococcus mutans.

The Inhibitory Ability of Cocoa Pod Husk Extract on Enterococcus faecalis Glucosyltransferase Enzyme Activity

The presence of Enterococcus faecalis in root canal is considered as one of the factors causing root canal treatment failure since the bacteria are capable of producing glucosyltransferase enzymes that play a role in forming endodontic biofilms. Hence, the bacteria are resistant to antibiotics. On the other hand, cocoa pod husk extract which is rich in chemical components especially flavonoids, tannins, and saponins, is thought to have an ability to inhibit Enterococcus faecalis glucosyltransferase enzyme activity. The aim of this research is to analyze the inhibitory ability of cocoa pod husk extract against E. faecalis glucosyltransferase enzyme activity. A total of 27 research samples were divided into three groups, namely, positive control (chlorhexidine gluconate 2%), negative control (aquades), and cocoa pod husk extract 3.12%. Next the enzymatic activity of each sample group was calculated based on the size of the fructose area read by high-performance liquid chromatography (HPLC) expressed in percent (%) and then converted to μmol/mL fructose which was considered as 1 unit of glucosyltransferase enzyme activity. Subsequently, the data were analyzed statistically using Kruskal-Wallis test. The results of data analysis using the Kruskal-Wallis test showed significant differences between groups of samples (p <0.05). Cocoa pod husk extract of 3.12% has inhibitory effect on E. faecalis glucosyltransferase enzyme activity. The use of cocoa pod husk extract meets the requirements and is proven useful as an irrigation agent in the treatment of root canals, because it contains antibacterial properties against E. faecalis.

Daya Hambat Aktivitas Enzim Glukosiltransferase (Gtf) Streptococcus mutans Oleh Ekstrak Temulawak (Curcuma Xanthorrhiza Roxb.)

Background: Streptococcus mutans is a bacteria which has glucosyltransferase (GTF) enzyme and acts as the main agent that causes dental caries. GTF enzyme will convert sucrose into fructose and glucan. Temulawak (Curcuma xanthorrhiza Roxb.) is one of the traditional herbs which has xanthorrhizol, curcumin, flavonoid, tanin, and saponin as an antibacterial agent. Purpose: The purpose of this research is to investigate the effect of temulawak extract (Curcuma xanthorrhiza Roxb.) to the activity of GTF enzyme Streptococcus mutans. Method: This research used 25%, 37,5%, and 50% concentration of temulawak extract as the treatment, and 0.12% chlorhexidine gluconate as a control. The method of this research consists of three steps; preparing the temulawak extract concentration of 25%, 37,5%, and 50%, preparing the GTF enzyme from the supernatant of Streptococcus mutans, and testing GTF enzyme activity by analyzing the fructose concentration using High Performance Liquid Chromatography (HPLC). Perusal of the fructose area was based on the retention time of fructose. One unit of GTF enzyme activity is defined as the 1 μmol fructose / ml of enzyme / hour. Result: The obtained data then were analyzed by Post-Hoc Tukey (HSD). The result showed a significant difference between each treatment group with the control group (p&lt;0.05). Conclussion: This research concludes that temulawak extract with 25%, 37,5%, and 50% concentrationcan’t inhibit the GTF enzyme activity of Streptococcus mutans.

The inhibition of Streptococcus mutans glucosyltransferase enzyme activity by mangosteen pericarp extract

Background: Streptococcus mutans (S. mutans) is a bacterium that plays an important role in the pathogenesis of dental caries. Streptococcus mutans produces the glucosyltransferase enzyme which is capable of catalyzing glucan synthesis in the progression of dental caries. Certain treatments involving traditional plant use have been developed to eradicate Streptococcus mutans as a means of preventing the formation of dental caries. One of these is mangosteen pericarp extract containing a number of polyphenols that have the capacity to act as antibacterial agents, namely; tannin, mangostin, and flavonoid. Purpose: The research aimed to investigate the inhibitory power of mangosteen pericarp extract against Streptococcus mutans producing the glucosyltransferase enzyme. Methods: The research used mangosteen pericarp extract at concentrations of 0.39% and 0.78% as the treatments, while 0.12% chlorhexidine gluconate was used as a positive control, and distilled water as a negative control. Each group consisted of six samples. Mangosteen peels extracted with 96% ethanol (maceration method) and mangosteen extract constituted 5% of the total weight of the mangosteen pericarp. Supernatant containing Gtf enzyme produced from a culture medium and centrifuged at 1500 rpm for 10 minutes at 4o C. Glucosyltransferase enzyme activity was measured by analyzing the extensive fructose area by means of High Performance Liquid Chromatography (HPLC). The extensive fructose area was determined according to time retention in each group. Results: Mangosteen peel extract at concentrations of 0.39% and 0.78% demonstrated greater ability than the negative control group (sterile aquades) and similar ability to the positive group (chlorhexidine 0.12%) to inhibit the activity of the Gtf enzyme or S. mutans bacteria. Conclusion: Mangosteen pericarp extract has the ability to inhibit the activity of Streptococcus mutans in producing glucosyltransferase enzyme.

EFEKTIVITAS BERKUMUR LARUTAN INFUSUM DAUN GUAVA (Psidium Guajava Linn) DALAM MENURUNKAN INDEKS PLAK

Plaque is a sticky deposit, consists of bacteria and it’s products in the form of biofilm that breed in a matrix on the surface of the tooth. Plaque can control chemically by using mouthwash. The infuse solution of guava leaf (Psidium guajava Linn) can be used as a herbal mouthwash. Antibacterial potency from the flavonoid in guava leaf can inhibit the growth of Streptococcus mutans and glucosyltransferase enzyme activity. The purpose of this studi is to determining the effectiveness of gargling guava leaf (Psidium guajava Linn) infuse solution in reducing plaque index. This study was conducted with pre-test post-test control group design to 32 students of Dentistry Faculty of Andalas University as respondents in two days. The first day, 32 respondents gargled with aquades as a control group and the second day, 32 respondents gargled with infuse solution of guava leaf (Psidium guajava Linn) with 80% concentration as the case group. The plaque index was observed with modified plaque index Turesky-Gilmore-Glicman of Quigley-Hein. Result of the study is there is difference in the reduction of plaque index between the control group and the case group significantly (p &lt;0.05), which the plaque index is lower in the case group than the control group. Conclusion of this study concluded that the infuse solution of guava leaf (Psidium guajava Linn) was effective in decreasing plaque index. &#x0D; Keywords:Infuse solution of Guava leaf (Psidium guajava Linn), plaque index

In vitro study of the effect of a lactoperoxidase-peroxidase-thiocyanate system with iodine on the cariogenicinity of streptococcus mutans

This study aimed to investigate the effect of a lactoperoxidase-peroxidase-thiocyanate (LPO-H2O-SCN-) system with different concentrations of iodine (I-) on Streptococcus mutans (S. mutans), particularly on various parameters, including growth, adhesion, glucosyltransferase (GTF) enzyme activity, and insoluble exopolysaccharide synthesis. S. mutans ATCC 25175 was used as experimental species. Clonal formation unit (CFU) were counted to investigate the inhibitory effect on bacterial growth. The inhibition rate of bacterial adherence was calculated to analyze the effect on adhesion. Anthrone method was used to determine the content of insoluble exopolysaccharides and the amount of reducing saccharides. GTF activity and enzyme activity were then determined. The inhibitory ability of the LPO-H2O2-SCN- system with I- on the cariogenicinity of S. mutans was strengthened as I- concentration was increased. At I- concentration > or = 100 micromol x L(-1) the antibacterial effects were significantly increased compared with those of the control group (P < 0.05). At I- concentration > or = 1,000 micromol x L(-1), the antibacterial effects were significantly improved compared with those of the group with SCN-only (P < 0.05). At I- concentration > or = 100 micromol x L(-1), the inhibition rate of bacterial adherence was > 50%; insoluble exopolysaccharide synthesis and GTF enzyme activity were reduced (P < 0.05). The antibacterial effects of the LPO-H2O2-I- system were enhanced by adding I- to overcome the antagonistic effect of physiological SCN- concentration. LPO-H2O2-SCN- system with different concentrations of I- showed statistically significant inhibitory effects on growth, adhesion, insoluble exopolysaccharide synthesis, and GTF enzyme activity.

Purification of Glucosyltransferase from Streptococcus sobrinus Cell Culture Medium by Combined Use of Batch Extraction and Countercurrent Chromatography with a Polymer Phase System

Glucosyltransferase (GTF), a cariogenic factor of Streptococcus sobrinus (SS), was extracted from a large quantity of cell culture medium by batch extraction with aqueous polymer two‐phase systems (APTPs). The small quantity of the extract was further purified by countercurrent chromatography (CCC) using polyethylene glycol (PEG) 8000‐dextran T500 system at pH 9.2, by eluting the impurities with the upper mobile phase at 1.0 mL/min. After the elution, the GTF still remaining in the dextran‐rich lower stationary phase was collected. The GTF fractions were subjected to hydroxyapatite chromatography to eliminate polymers, such as PEG 8000 and dextran T500. Purified fractions were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) and GTF enzyme activity, which rose over 270 fold from that in the original culture medium.

Modification of surface histidine residues abolishes the cytotoxic activity of Clostridium difficile toxin A

Clostridium difficile toxin A displays both cytotoxic and enterotoxic activities. It has recently been demonstrated that toxin A exerts its cytotoxic effect by the glucosylation of the small GTP-binding proteins of the Rho family. Diethyl pyrocarbonate, at pH 7.0, was used to chemically modify exposed histidine residues on toxin A. Modification of toxin A with diethyl pyrocarbonate abolished both its cytotoxic activity and the ability of the toxin to bind Zn-Sepharose gel. Treatment of toxin A with [ 14C]-diethyl pyrocarbonate revealed concentration dependent labelling of histidine residues on the toxin molecules. The effects of diethyl pyrocarbonate could be reversed by hydroxylamine treatment. These data suggest the modified histidine residues on toxin A are critical to its cytotoxic activity. Histidine modification had no effect on the glucosyl transferase enzyme activity of toxin A. However, modification abolished the ‘cold’ binding of toxin to bovine thyroglobulin in an ELISA and reduced ligand binding activity in a rabbit erythrocyte haemagglutination assay. The data suggest that the histidine residues may be crucial to the receptor-binding activity of toxin A. Exposed histidines on toxin A are available for zinc chelation, and these have been exploited in the development of a novel purification protocol for toxin A using zinc-chelating chromatography.

Dextran-induced aggregation in a mutant of Streptococcus sobrinus 6715-13.

A mutant of wild-type Streptococcus sobrinus 6715-13 has been isolated which resists aggregation by exogenous dextran. This variant is able to form adherent plaque deposits in vitro when cultured in the presence of sucrose and has dextranase activity. In these respects it is the complement of previously described isolates which are plaque formation defective but aggregation normal. Measurements of the incorporation of glucose from glucosyl-labeled sucrose into glucan by cell-associated glucosyltransferase enzyme activity and the thermal labilities of catalytic and receptor functions, as well as the binding of labeled dextrans to the cells, provide evidence that neither dextranase nor glucosyltransferase is the receptor involved in dextran-induced aggregation. Blockage of such bacterial aggregation by anti-glucosyltransferase or anti-dextranase sera suggests cross-reactivity between the antigenic determinants of proteins which recognize alpha(1-6) glucan linkages. A model is proposed, consistent with these and previous findings, in which enzymatic function precedes dextran receptor activity in emergence from the cell. It is also proposed that dextran receptor components of the multireactive glucosyltransferase enzyme(s) and dextranase(s) are spatially separate from, although functionally and antigenically related to, the receptors on the bacterial surface involved in dextran-induced aggregation.

Interaction of soluble glucosyl- and mannosyl-transferase enzyme activities in the synthesis of a glucomannan

A neutral-detergent-solubilized-enzyme preparation derived from Phaseolus aureus hypocotyls contains two types of glycosyltransferase activity. One, mannosyltransferase enzyme activity, utilizes GDP-alpha-d-mannose as the sugar nucleotide substrate. The other, glucosyltransferase enzyme activity, utilizes GDP-alpha-d-glucose as the sugar nucleotide substrate. The soluble enzyme preparation catalyses the formation of what appears to be a homopolysaccharide when either sugar nucleotide is the only substrate present. A beta-(1-->4)-linked mannan is the only polymeric product when only GDP-alpha-d-mannose is added. A beta-(1-->4)-linked glucan is the only polymeric product when only GDP-alpha-d-glucose is added. In the presence of both sugar nucleotides, however, a beta-(1-->4)-linked glucomannan is formed. There are indications that endogenous sugar donors may be present in the enzyme preparation. There appear to be only two glycosyltransferases in the enzyme preparation, each catalysing the transfer of a different sugar to the same type of acceptor molecule. The glucosyltransferase requires the continual production of mannose-containing acceptor molecules for maintenance of enzyme activity, and is thereby dependent upon the activity of the mannosyltransferase. The mannosyltransferase, on the other hand, does not require the continual production of glucose-containing acceptors for maintenance of enzyme activity, but is severely inhibited by GDP-alpha-P-glucose. These properties promote the synthesis of beta-(1-->4)-linked glucomannan rather than beta-(1-->4)-linked glucan plus beta-(1-->4)-linked mannan when both sugar nucleotide substrates are present.