==================================BSR29================================== 29. Sugar transport in lactic and oral streptococci: regulation of; mechanism of; phosphoenolpyruvate hexose phosphotransferases (PTS); phosphoenolpyruvate sugar phosphotransferase system; sugar uptake, accumulation; energy coupling, role of energy; metabolism. Streptococcus lactis; S. cremoris; S. diacetylactis; S. thermophilus; S. mutans; S. salivarius; S. sanguis. 1 UI - 87099170 AU - Keevil CW ; McDermid AS ; Marsh PD ; Ellwood DC TI - Protonmotive force driven 6-deoxyglucose uptake by the oral pathogen, Streptococcus mutans Ingbritt. AB - Streptococcus mutans Ingbritt was grown in glucose-excess continuous culture to repress the glucose phosphoenolpyruvate phosphotransferase system (PTS) and allow investigation of the alternative glucose process using the non-PTS substrate, (3H) 6-deoxyglucose. After correcting for non-specific adsorption to inactivated cells, the radiolabelled glucose analogue was found to be concentrated approximately 4.3-fold intracellularly by bacteria incubated in 100 mM Tris-citrate buffer, pH 7.0. Mercaptoethanol or KCl enhanced 6-deoxyglucose uptake, enabling it to be concentrated internally by at least 8-fold, but NaCl was inhibitory to its transport. Initial uptake was antagonised by glucose but not 2-deoxyglucose. Evidence that 6-deoxyglucose transport was driven by protonmotive force (delta p) was obtained by inhibiting its uptake with the protonophores, 2,4-dinitrophenol, carbonylcyanide m-chlorophenylhydrazine, gramicidin and nigericin, and the electrical potential difference (delta psi) dissipator, KSCN. The membrane ATPase inhibitor, N,N1-dicyclohexyl carbodiimide, also reduced 6-deoxyglucose uptake as did 100 mM lactate. In combination, these two inhibitors completely abolished 6-deoxyglucose transport. This suggests that the driving force for 6-deoxyglucose uptake is electrogenic, involving both the transmembrane pH gradient (delta pH) and delta psi. ATP hydrolysis, catalysed by the ATPase, and lactate excretion might be important contributors to delta pH. MH - Deoxy Sugars/*METABOLISM ; Deoxyglucose/*METABOLISM ; Glucose/METABOLISM ; Heat ; Human ; Hydrogen-Ion Concentration ; Ionophores ; Mercaptoethanol ; Potassium Chloride ; Sodium Chloride ; Streptococcus mutans/*METABOLISM ; Support, Non-U.S. Gov't SO - Arch Microbiol 1986 Nov;146(2):118-24 2 UI - 87076587 AU - Deutscher J ; Pevec B ; Beyreuther K ; Kiltz HH ; Hengstenberg W TI - Streptococcal phosphoenolpyruvate-sugar phosphotransferase system: amino acid sequence and site of ATP-dependent phosphorylation of HPr. AB - The amino acid sequence of histidine-containing protein (HPr) from Streptococcus faecalis has been determined by direct Edman degradation of intact HPr and by amino acid sequence analysis of tryptic peptides, V8 proteolytic peptides, thermolytic peptides, and cyanogen bromide cleavage products. HPr from S. faecalis was found to contain 89 amino acid residues, corresponding to a molecular weight of 9438. The amino acid sequence of HPr from S. faecalis shows extended homology to the primary structure of HPr proteins from other bacteria. Besides the phosphoenolpyruvate-dependent phosphorylation of a histidyl residue in HPr, catalyzed by enzyme I of the bacterial phosphotransferase system, HPr was also found to be phosphorylated at a seryl residue in an ATP-dependent protein kinase catalyzed reaction [Deutscher, J., & Saier, M. H., Jr. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 6790-6794]. The site of ATP-dependent phosphorylation in HPr of S. faecalis has now been determined. [32P]P-Ser-HPr was digested with three different proteases, and in each case, a single labeled peptide was isolated. Following digestion with subtilisin, we obtained a peptide with the sequence -(P)Ser-Ile-Met-. Using chymotrypsin, we isolated a peptide with the sequence -Ser-Val-Asn-Leu-Lys-(P)Ser-Ile-Met-Gly-Val-Met-. The longest labeled peptide was obtained with V8 staphylococcal protease. According to amino acid analysis, this peptide contained 36 out of the 89 amino acid residues of HPr. The following sequence of 12 amino acid residues of the V8 peptide was determined: -Tyr-Lys-Gly-Lys-Ser-Val-Asn-Leu-Lys-(P)Ser-Ile-Met-.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Adenosine Triphosphate/METABOLISM ; Amino Acid Sequence ; Bacterial Proteins/ISOLATION & PURIFICATION/*METABOLISM ; Binding Sites ; Peptide Fragments/ANALYSIS ; Phosphoenolpyruvate Sugar Phosphotransferase System/ ISOLATION & PURIFICATION/*METABOLISM ; Phosphorylation ; Streptococcus Faecalis/*ENZYMOLOGY ; Support, Non-U.S. Gov't SO - Biochemistry 1986 Oct 21;25(21):6543-51 3 UI - 87074865 AU - Martin SA ; Russell JB TI - Phosphoenolpyruvate-dependent phosphorylation of hexoses by ruminal bacteria: evidence for the phosphotransferase transport system. AB - Six species of ruminal bacteria were surveyed for the phosphoenolpyruvate (PEP)-dependent phosphorylation of glucose. Selenomonas ruminantium HD4, Streptococcus bovis JB1, and Megasphaera elsdenii B159 all showed significant activity, but Butyrivibrio fibrisolvens 49, Bacteroides succinogenes S85, and Bacteroides ruminicola B1(4) showed low rates of PEP-dependent phosphorylation and much higher rates in the presence of ATP. S. ruminantium HD4, S. bovis JB1, and M. elsdenii B159 also used PEP to phosphorylate the nonmetabolizable glucose analog 2-deoxy-D-glucose (2-DG). Rates of 2-DG phosphorylation with ATP were negligible for S. bovis JB1 and M. elsdenii B159, but toluene-treated cells of S. ruminantium HD4 phosphorylated 2-DG in the presence of ATP as well as PEP. Cell-free extracts of S. ruminantium HD4 used ATP but not PEP to phosphorylate glucose and 2-DG. Since PEP could serve as a phosphoryl donor in toluene-treated cells but not in cell-free extracts, there was evidence for membrane and hence phosphotransferase system involvement in the PEP-dependent activity. The ATP-dependent phosphorylating enzymes from S. ruminantium HD4 and S. bovis JB1 had molecular weights of approximately 48,000 and were not inhibited by glucose 6-phosphate. Based on these criteria, they were glucokinases rather than hexokinases. The S. ruminantium HD4 glucokinase was competitively inhibited by 2-DG and mannose, sugars that differ from glucose in the C-2 position. Since 2-DG was a competitive inhibitor of glucose, the same enzyme probably phosphorylates both sugars. The S. bovis JB1 glucokinase was not inhibited by either 2-DG or mannose and had a higher Km and Vmax for glucose. MH - Adenosine Triphosphate/METABOLISM ; Animal ; Bacteria/*ENZYMOLOGY ; Bacteroidaceae/ENZYMOLOGY ; Bacteroides/ENZYMOLOGY ; Deoxyglucose/ METABOLISM ; Glucokinase/METABOLISM ; Glucose/*METABOLISM ; Phosphoenolpyruvate Sugar Phosphotransferase System/*METABOLISM ; Phosphoenolpyruvate/METABOLISM ; Phosphorylation ; Rumen/*MICROBIOLOGY ; Streptococcus/ENZYMOLOGY ; Support, Non-U.S. Gov't ; Veillonellaceae/ ENZYMOLOGY SO - Appl Environ Microbiol 1986 Dec;52(6):1348-52 4 UI - 87008430 AU - Ehrenfeld EE ; Kessler RE ; Clewell DB TI - Identification of pheromone-induced surface proteins in Streptococcus faecalis and evidence of a role for lipoteichoic acid in formation of mating aggregates. AB - The conjugative transfer of the Streptococcus faecalis plasmid pAD1 is characterized by a 10,000-fold increase in frequency following sex pheromone (cAD1) induction. Before the increase in plasmid transfer, donor cells synthesize a proteinaceous adhesin that facilitates the formation of mating aggregates. Four novel surface proteins appearing after exposure of pAD1-containing cells to sex pheromone have been identified. Thirty minutes after induction, a 130-kilodalton (kDa) protein was detectable by Western blotting. A 74-kDa protein, the major species present, and a pair of bands at 153 and 157 kDa were evident 45 min after induction. Induced cells containing another conjugative S. faecalis plasmid, pPD1, gave rise to three high-molecular-weight proteins of the same size (130, 153, and 157 kDa) as those synthesized by pAD1-containing cells. These proteins cross-reacted with antisera raised against induced cells containing pAD1. However, the major protein species produced by pPD1-containing cells had a molecular weight of 78,000 and did not cross-react significantly with the corresponding band of the pAD1 system. Pheromone-induced transfer of the two plasmids, when both were present in the same cell, was independent; induction was limited to the pheromone-specified plasmid. The possibility that lipoteichoic acid might act as a receptor (binding substance) for the induced adhesin protein was also explored. Free lipoteichoic acid (isolated from S. faecalis) inhibited clumping of induced cells, apparently by acting as a competitive inhibitor of the cellular binding substance. MH - Bacterial Proteins/BIOSYNTHESIS ; *Conjugation, Genetic ; Membrane Proteins/*BIOSYNTHESIS ; Pheromones/*PHARMACODYNAMICS ; Phosphatidic Acids/*METABOLISM/PHARMACODYNAMICS ; Plasmids ; Streptococcus Faecalis/ GENETICS/*METABOLISM ; Support, U.S. Gov't, P.H.S. ; Teichoic Acids/ *METABOLISM/PHARMACODYNAMICS SO - J Bacteriol 1986 Oct;168(1):6-12 5 UI - 87008072 AU - Marounek M ; Bartos S TI - Stoichiometry of glucose and starch splitting by strains of amylolytic bacteria from the rumen and anaerobic digester. AB - The stoichiometry of glucose and starch splitting by the amylolytic bacteria Streptococcus bovis, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Eubacterium ruminantium and Clostridium sp. was followed. There were many differences in the ratios of metabolites and in growth yields, as well as in the cell composition, between the growth on glucose and starch. The bacteria employ different nutritional strategies with respect to both energy sources. MH - Animal ; Bacteria, Anaerobic/GROWTH & DEVELOPMENT/*METABOLISM ; Cattle ; Clostridium/GROWTH & DEVELOPMENT/METABOLISM ; Culture Media ; Eubacterium/ GROWTH & DEVELOPMENT/METABOLISM ; Fermentation ; Glucose/*METABOLISM ; Gram-Negative Anaerobic Bacteria/GROWTH & DEVELOPMENT/METABOLISM ; Rumen/ MICROBIOLOGY ; Sheep ; Starch/*METABOLISM ; Streptococcus/GROWTH & DEVELOPMENT/METABOLISM SO - J Appl Bacteriol 1986 Jul;61(1):81-6 6 UI - 86319188 AU - Assev S ; Scheie AA TI - Xylitol metabolism in xylitol-sensitive and xylitol-resistant strains of streptococci. AB - 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. MH - Drug Resistance, Microbial ; Fructose/METABOLISM ; Streptococcus/DRUG EFFECTS/GROWTH & DEVELOPMENT/*METABOLISM ; Streptococcus mutans ; Streptococcus sanguis ; Xylitol/*METABOLISM SO - Acta Pathol Microbiol Immunol Scand [B] 1986 Aug;94(4):239-43 7 UI - 86304176 AU - Inamine JM ; Lee LN ; LeBlanc DJ TI - Molecular and genetic characterization of lactose-metabolic genes of Streptococcus cremoris. AB - Lac+ plasmid DNA from Streptococcus cremoris H2 was subcloned with an Escherichia coli vector on a 3.5-kilobase-pair PstI-AvaI fragment. Genetic analysis of the cloned DNA was possible because linear Lac+ DNA fragments were productive in the S. sanguis transformation system. Complementation of S. sanguis Lac-mutants showed that the 3.5-kilobase-pair fragment included the structural gene for 6-phospho-beta-D-galactosidase and either enzyme II-lac or factor III-lac of the lactose-specific phosphoenolpyruvate-dependent phosphotransferase system. Expression of the S. cremoris-like 40,000-dalton 6-phospho-beta-D-galactosidase in S. sanguis Lac+ transformants, rather than the 52,000-dalton wild-type S. sanguis enzyme, demonstrated the occurrence of gene replacement and not gene repair. The evidence supports chromosomal integration as the mechanism by which S. sanguis Lac- recipients are converted to a Lac+ phenotype after transformation with Lac+ DNA. Southern blot data suggest that the Lac+ DNA does not reside on a transposon, but that integration always occurs within a specific HincII fragment of the recipient chromosome. Hybridization experiments demonstrate homology between the S. cremoris Lac+ DNA and cellular DNA from Lac+ strains of Streptococcus lactis, S. mutans, S. faecalis, and S. sanguis. MH - Bacterial Proteins/*GENETICS ; Beta-Galactosidases/*GENETICS ; Comparative Study ; Escherichia Coli/GENETICS ; Galactosidases/*GENETICS ; *Genes, Bacterial ; Genes, Structural ; Phosphoenolpyruvate Sugar Phosphotransferase System/*GENETICS ; Plasmids ; Recombinant Proteins/ GENETICS ; Sequence Homology, Nucleic Acid ; Species Specificity ; Streptococcus/*GENETICS/METABOLISM ; Support, U.S. Gov't, P.H.S. ; Transformation, Genetic SO - J Bacteriol 1986 Sep;167(3):855-62 8 UI - 86303021 AU - Aoki H ; Shiroza T ; Hayakawa M ; Sato S ; Kuramitsu HK TI - Cloning of a Streptococcus mutans glucosyltransferase gene coding for insoluble glucan synthesis [published erratum appears in Infect Immun 1986 Dec;54(3):931] AB - The gtfB gene coding for a glucosyltransferase (GTF) activity of Streptococcus mutans GS-5 was isolated on a 15.4-kilobase DNA fragment by using a lambda L47.1 gene library. The activity was catalyzed by gene products of 150 and 145 kilodaltons which reacted with antibodies directed against both soluble and insoluble glucan-synthesizing GTFs. The enzyme present in crude Escherichia coli extracts synthesized both soluble and insoluble glucans. The enzyme was partially purified from lysates of the lambda DS-76 clone and synthesized both types of glucans in a primer-independent fashion. In addition, the purified enzyme exhibited a pI of approximately 5.0. Southern blot analysis indicated that the cloned GTF gene represented a contiguous nucleotide sequence on the strain GS-5 chromosome. Furthermore, evidence for the existence of a distinct gene sharing partial homology with gtfB was also obtained. The gtfB gene was subcloned into plasmid pACYC184 into E. coli and exhibited GTF activity when carried on GS-5 inserts as small as 5 kilobases. The approximate location of the GTF promoter and the direction of gene transcription were also determined. The cloned enzyme was not secreted through the cytoplasmic membrane of E. coli, since most of the activity was found in the cytoplasm and, in lesser amounts, associated with the cytoplasmic membrane. The gtfB gene was insertionally inactivated by introducing a gene fragment coding for erythromycin resistance into the GTF coding region. After transformation of strain GS-5 with the altered gene, transformants defective in insoluble glucan synthesis were identified. These results indicate that the gtfB gene codes for a GTF involved in insoluble glucan synthesis in strain GS-5. MH - Base Sequence ; *Cloning, Molecular ; DNA Insertion Elements ; Electrophoresis, Polyacrylamide Gel ; Escherichia Coli/ENZYMOLOGY ; *Genes, Bacterial ; Glucans/*BIOSYNTHESIS ; Glucosyltransferases/ *GENETICS/ISOLATION & PURIFICATION ; Streptococcus mutans/ENZYMOLOGY/ *GENETICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Infect Immun 1986 Sep;53(3):587-94 9 UI - 86267539 AU - Germaine GR ; Tellefson LM TI - Role of the cell membrane in pH-dependent fluoride inhibition of glucose uptake by Streptococcus mutans. AB - The effect of pH on the fluoride sensitivity of glucose uptake by whole cells and glucose transport by permeabilized cells of Streptococcus mutans was compared. Whole cells exhibited a marked pH-dependent sensitivity to fluoride over the pH range 7.0 to 5.0. As the pH was decreased, fluoride sensitivity increased. In contrast, no significant effect of pH on the fluoride sensitivity of glucose transport (e.g., phosphorylation) by permeabilized cells energized with 2-phosphoglycerate was noted. The relative effect of pH on the fluoride sensitivity of whole cell glucose uptake and fermentation was similar. These data are consistent with the notion that the cell membrane is impermeable to the fluoride anion and that intracellular accumulation of fluoride depends on translocation of hydrogen fluoride across the membrane. MH - Cell Membrane/METABOLISM ; Fermentation ; Fluorides/*PHARMACODYNAMICS ; Glucose/*METABOLISM ; Hydrogen-Ion Concentration ; Sodium Fluoride/ PHARMACODYNAMICS ; Streptococcus mutans/DRUG EFFECTS/*METABOLISM ; Support, U.S. Gov't, P.H.S. SO - Antimicrob Agents Chemother 1986 Jan;29(1):58-61 10 UI - 86244528 AU - Waygood EB ; Mattoo RL ; Erickson E ; Vadeboncoeur C TI - Phosphoproteins and the phosphoenolpyruvate:sugar phosphotransferase system of Streptococcus salivarius. Detection of two different ATP-dependent phosphorylations of the phosphocarrier protein HPr. AB - Phosphoproteins which arise from incubation of Streptococcus salivarius ATCC25975 crude extracts with [32P]phosphoenolpyruvate and [gamma-32P]ATP, were separated and detected by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and autoradiography. These procedures were carried out using the methodology that has been developed to allow for the detection of phosphoproteins containing 1-P-histidinyl and 3-P-histidinyl residues, and also to distinguish between these and phosphoproteins containing acid-stable phosphoamino acids such as phosphoserine, phosphothreonine, and phosphotyrosine. Extracts of cells which had been grown with various sugars as carbon sources were investigated to determine both constitutive and inducible phosphoproteins. No evidence was found for phosphoproteins specifically induced by a sugar, and in particular no evidence was found for any IIIsugar phosphocarrier protein of the phosphoenolpyruvate:sugar phosphotransferase system (PTS). Incubation with [gamma-32P]ATP showed that histidine-containing phosphocarrier protein (HPr) of the PTS could be phosphorylated to give both acid-stable and acid-labile phosphoamino acid residues. The acid-labile ATP-dependent phosphorylation activity was activated by glucose-6-P and appeared to produce a 3-P-histidinyl residue in HPr. MH - Adenosine Triphosphate/METABOLISM ; Autoradiography ; Bacterial Proteins/ *METABOLISM ; Electrophoresis, Polyacrylamide Gel ; Phosphoenolpyruvate/ METABOLISM ; Phosphoenolpyruvate Sugar Phosphotransferase System/ *METABOLISM ; Phosphoproteins/*METABOLISM ; Phosphorylation ; Streptococcus/*ENZYMOLOGY ; Support, Non-U.S. Gov't SO - Can J Microbiol 1986 Apr;32(4):310-8 11 UI - 86224845 AU - Zero DT ; van Houte J ; Russo J TI - The intra-oral effect on enamel demineralization of extracellular matrix material synthesized from sucrose by Streptococcus mutans. AB - The role of extracellular matrix material (EMM) synthesized from sucrose (S) by Streptococcus mutans IB-1600 in altering the demineralizing potential of artificial plaque was evaluated with an intraoral enamel demineralization test (IEDT). The artificial plaque samples were prepared from cells cultivated in Todd-Hewitt broth (THB) supplemented with various S concentrations and by mixing THB-grown cells with increasing proportions of EMM (heat-killed THB + 2% S-cultivated cells). The samples were also evaluated for cell density (DNA content) and acidogenicity in vitro (pH-stat), as well as for in situ pH changes during a 45-minute intra-oral test following a 10% glucose rinse. An increase in the proportion of EMM relative to cell density was associated with an increase in enamel demineralization. This trend reversed when the ratio of cells to EMM was less than 1:19. Experiments involving strains of S. mitis, S. sanguis, and S. salivarius suggested a similar effect of EMM. The intra-oral pH data suggest that the presence of EMM may enhance demineralization by altering diffusion properties of plaque. MH - Acids/METABOLISM ; Adult ; Animal ; Cattle ; Dental Caries/MICROBIOLOGY/ *PHYSIOPATHOLOGY ; Dental Enamel/*DRUG EFFECTS ; Dental Plaque/ MICROBIOLOGY ; DNA, Bacterial/ANALYSIS ; Extracellular Matrix/*METABOLISM ; Glucans/BIOSYNTHESIS ; Glucose/METABOLISM/PHARMACODYNAMICS ; Human ; Hydrogen-Ion Concentration ; Male ; Middle Age ; Polysaccharides, Bacterial/BIOSYNTHESIS/*PHARMACODYNAMICS ; Streptococcus mutans/ANALYSIS/ *METABOLISM ; Sucrose/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Dent Res 1986 Jun;65(6):918-23 12 UI - 86223800 AU - Deutscher J ; Sauerwald H TI - Stimulation of dihydroxyacetone and glycerol kinase activity in Streptococcus faecalis by phosphoenolpyruvate-dependent phosphorylation catalyzed by enzyme I and HPr of the phosphotransferase system. AB - Recently we reported the phosphoenolpyruvate (PEP)-dependent phosphorylation of a 55-kilodalton protein of Streptococcus faecalis catalyzed by enzyme I and histidine-containing protein (HPr) of the phosphotransferase system (J. Deutscher, FEMS Microbiol. Lett. 29:237-243, 1985). The purified 55-kilodalton protein was found to exhibit dihydroxyacetone kinase activity. Glycerol was six times more slowly phosphorylated than dihydroxyacetone. The Kms were found to be 0.7 mM for ATP, 0.45 mM for dihydroxyacetone, and 0.9 mM for glycerol. PEP-dependent phosphorylation of dihydroxyacetone kinase stimulated phosphorylation of both substrates about 10-fold. Fructose 1,6-diphosphate at concentrations higher than 2 mM inhibited the activity of phosphorylated and unphosphorylated dihydroxyacetone kinase in a noncompetitive manner. The rate of PEP-dependent phosphorylation of dihydroxyacetone kinase was about 200-fold slower than the phosphorylation rate of III proteins (also called enzyme III or factor III), which so far have been considered the only phosphoryl acceptors of histidyl-phosphorylated HPr. P-Dihydroxyacetone kinase was found to be able to transfer its phosphoryl group in a backward reaction to HPr. Following [32P]PEP-dependent phosphorylation and tryptic digestion of dihydroxyacetone kinase, we isolated a labeled peptide composed of 37 amino acids, as determined by amino acid analysis. The single histidyl residue of this peptide most likely carries the phosphoryl group in phosphorylated dihydroxyacetone kinase. MH - Chromatography, High Pressure Liquid ; Fructosediphosphates/ PHARMACODYNAMICS ; Glycerol Kinase/*METABOLISM ; Kinetics ; Molecular Weight ; Peptide Fragments/ISOLATION & PURIFICATION ; Phosphoenolpyruvate/ *METABOLISM ; Phosphoenolpyruvate Sugar Phosphotransferase System/ *METABOLISM ; Phosphorylation ; Phosphotransferases, ATP/*METABOLISM ; Streptococcus Faecalis/*ENZYMOLOGY ; Substrate Specificity ; Trypsin/ METABOLISM SO - J Bacteriol 1986 Jun;166(3):829-36 13 UI - 86207904 AU - Wennerholm K ; Emilson CG ; Krasse B TI - Oral glucose clearance in subjects with high or low salivary levels of Streptococcus mutans and lactobacilli. AB - The ability to eliminate sugar from the oral cavity was studied in subjects with high or low numbers of salivary Streptococcus mutans and lactobacilli. The glucose concentration in saliva was determined after consumption of biscuits using the Gloxtest, and the logarithms of the glucose values were plotted against time. The sugar clearance time was measured where the slope of the plot intersected with a glucose level of 1 mg/ml. Subjects with a high level of S. mutans had a significantly longer clearance time than subjects with a low level. More salivary lactobacilli were found in a subject group with a long sugar clearance time than in a group with a fast clearance rate. MH - Adolescence ; Adult ; Glucose/ADMINISTRATION & DOSAGE/*METABOLISM ; Human ; Lactobacillus/*ISOLATION & PURIFICATION/METABOLISM ; Saliva/*METABOLISM/ MICROBIOLOGY ; Streptococcus mutans/*ISOLATION & PURIFICATION/METABOLISM ; Support, Non-U.S. Gov't ; Time Factors SO - Scand J Dent Res 1986 Apr;94(2):121-4 14 UI - 86196052 AU - Brissette JL ; Cabacungan EA ; Pieringer RA TI - Studies on the antibacterial activity of dodecylglycerol. Its limited metabolism and inhibition of glycerolipid and lipoteichoic acid biosynthesis in Streptococcus mutans BHT. AB - Growth-inhibitory concentrations of racemic sn-1(3)-dodecylglycerol inhibit the incorporation of [14C] glycerol into lipids and lipoteichoic acid of Streptococcus mutans BHT and alter the per cent composition of the glycerolipids. Increases in phosphatidic acid and diphosphatidylglycerol (at the expense of phosphatidylglycerol) contribute the most to the change in lipid composition. No cellular lysis occurs under these conditions. Radioactive racemic sn-1(3)-dodecylglycerol is readily taken up by the cell and is metabolized primarily to lysophosphatidic acid and phosphatidic acid with smaller amounts converted to phosphatidylglycerol and diacylglycerol. The accumulation of phosphatidic acid and the loss of viability respond in parallel to different concentrations of dodecylglycerol. An increase in CTP is also observed which together with the increase in phosphatidic acid suggests a possible impairment in the synthesis of CDP-diacylglycerol. MH - Alkaline Phosphatase/METABOLISM ; Glycerides/METABOLISM/*PHARMACODYNAMICS ; Glycerin/*ANALOGS & DERIVATIVES/BIOSYNTHESIS ; Laurates/METABOLISM/ *PHARMACODYNAMICS ; Lauric Acids/*PHARMACODYNAMICS ; Lipids/*BIOSYNTHESIS ; Models, Chemical ; Nucleotides/ANALYSIS ; Phosphatidic Acids/ANALYSIS/ *BIOSYNTHESIS ; Phosphatidylglycerols/ANALYSIS ; Streptococcus mutans/ *METABOLISM ; Support, U.S. Gov't, P.H.S. ; Teichoic Acids/*BIOSYNTHESIS SO - J Biol Chem 1986 May 15;261(14):6338-45 15 UI - 86196003 AU - Triscott MX ; van de Rijn I TI - Solubilization of hyaluronic acid synthetic activity from streptococci and its activation with phospholipids. AB - To date all hyaluronic acid synthetic systems have been of a particulate nature, and attempts at solubilization have been unsuccessful. This has hampered attempts to elucidate the mechanism by which hyaluronic acid is produced. In this paper we demonstrate that the hyaluronic acid synthetic activity from group C streptococcal membranes was solubilized using 2% digitonin and that the activity was optimized by reconstitution with cardiolipin at an optimum phospholipid/protein ratio (microgram/microgram) of 5:1. Furthermore, chromatography of the solubilized synthetase demonstrated that it eluted after the void volume of a Sepharose CL-6B column. CHAPSO, octyl glucopyranoside, sodium cholate, Triton X-100, and zwittergent 314 either inhibited or failed to solubilize the synthetic activity. Phospholipids other than cardiolipin also reconstituted the activity from the digitonin extract, particularly phosphatidylethanolamine and phosphatidylserine. In our system, the specific activity of hyaluronic acid synthetase was increased up to 63 times that of the system of the intact membrane. Furthermore, the total activity of the reconstituted system was 4.9 times greater than that of intact membranes. The soluble enzyme system showed similarities to the membrane-bound synthetase in the kinetics of production of trichloroacetic acid-soluble and -insoluble hyaluronic acid, and the hyaluronic acid produced was of comparable molecular weight. MH - Carbon Radioisotopes ; Cell Membrane/METABOLISM ; Detergents/ PHARMACODYNAMICS ; Digitonin/PHARMACODYNAMICS ; Hyaluronic Acid/ *BIOSYNTHESIS/ISOLATION & PURIFICATION ; Kinetics ; Phospholipids/ *PHARMACODYNAMICS ; Streptococcus/DRUG EFFECTS/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 May 5;261(13):6004-9 16 UI - 86194716 AU - Stimpson SA ; Esser RE ; Cromartie WJ ; Schwab JH TI - Comparison of in vivo degradation of 125I-labeled peptidoglycan-polysaccharide fragments from group A and group D streptococci. AB - The in vivo degradation and persistence of 125I-labeled peptidoglycan-polysaccharide (PG-PS) fragments from the cell walls of group A and D streptococci were compared by group after intraperitoneal injection into rats. The quantity of PG-PS in the livers and spleens of group D PG-PS-injected rats was less than the quantity in rats injected with group A PG-PS throughout the course of the experiment. Gel filtration analyses of liver and spleen homogenates indicated that group A PG-PS was relatively resistant to degradation, whereas group D PG-PS was extensively degraded to yield a heterogeneous mixture of fragments of lower molecular weight. There was no significant difference in the content of group A PG-PS versus that of group D in joints or blood samples. Analysis of fragment sizes in these tissues also indicated more extensive degradation of group D PG-PS. However, the majority of group A PG-PS in blood samples and joints was a lower molecular weight than that found in the livers or spleens. We conclude that group A PG-PS undergoes a significant but low level of degradation and that group D PG-PS is much less persistent and more extensively degraded than group A PG-PS is in vivo. These differences in PG-PS catabolism may account, in part, for the capacity of group A PG-PS to induce chronic, recurrent arthritis of longer duration than that induced by group D PG-PS. MH - Animal ; Cell Wall/METABOLISM ; Comparative Study ; Female ; Joints/ METABOLISM ; Liver/METABOLISM ; Metabolic Clearance Rate ; Molecular Weight ; Peptidoglycan/*METABOLISM ; Polysaccharides, Bacterial/ *METABOLISM ; Rats ; Species Specificity ; Spleen/METABOLISM ; Streptococcus/*METABOLISM ; Streptococcus Pyogenes/*METABOLISM ; Support, U.S. Gov't, P.H.S. ; Tissue Distribution SO - Infect Immun 1986 May;52(2):390-6 17 UI - 86189744 AU - Nisizawa T ; Takeuchi K ; Imai S ; Kitahata S ; Okada S TI - Difference in mode of inhibition between alpha-D-xylosyl beta-D-fructoside and alpha-isomaltosyl beta-D-fructoside in synthesis of glucan by Streptococcus mutans D-glucosyltransferase. AB - Both alpha-isomaltosyl beta-D-fructoside and alpha-D-xylosyl beta-D-fructoside show strong inhibition of the synthesis of water-insoluble and water-soluble D-glucans from sucrose by a partially purified preparation of a D-glucosyltransferase (GTase) from Streptococcus mutans 6715; however, the inhibitory modes differ substantially. In the presence of alpha-isomaltosyl beta-D-fructoside, the production of reducing sugars and the consumption of sucrose are remarkably enhanced, compared with a control of sucrose alone. Under these conditions, a large proportion of low-molecular-weight glycan (lmwg) and a series of nonreducing oligosaccharides (both containing D-fructosyl groups or residues) are produced. In contrast, in the presence of alpha-D-xylosyl beta-D-fructoside, the production of reducing sugars and the sucrose consumption are strikingly suppressed, and no lmwg or oligosaccharides are produced. Thus, it may be concluded that alpha-isomaltosyl beta-D-fructoside acts as an alternative acceptor for the D-glucosyl and/or D-glucanosyl transfer reactions of the enzyme, and serves to lessen the formation of insoluble and soluble D-glucan, although it stimulates the transferring activity of the enzyme. On the other hand, alpha-D-xylosyl beta-D-fructoside competitively inhibits the sucrose-splitting activity of the enzyme as an analog to sucrose, and thereby diminishes the synthesis of D-glucan. MH - Comparative Study ; Disaccharides/*PHARMACODYNAMICS ; Glucans/ *BIOSYNTHESIS ; Glucosyltransferases/*ANTAGONISTS & INHIBITORS/ISOLATION & PURIFICATION ; Kinetics ; Oligosaccharides/ANALYSIS ; Streptococcus mutans/*ENZYMOLOGY ; Structure-Activity Relationship ; Support, Non-U.S. Gov't ; Trisaccharides/*PHARMACODYNAMICS SO - Carbohydr Res 1986 Mar 1;147(1):135-44 18 UI - 86161499 AU - Walker GJ ; Schuerch C TI - Activity of branched dextrans in the acceptor reaction of a glucosyltransferase (GTF-I) from Streptococcus mutans OMZ176. AB - The ability of several native and chemically synthesized, branched dextrans to stimulate the activity of an alpha-D-glucosyltransferase (GTF-I) of Streptococcus mutans has been compared. The enzyme catalysed the transfer of glucosyl residues from sucrose with the formation of water-insoluble (1----3)-alpha-D-glucan. The rate of this reaction was greatly increased in the presence of dextran, and the extent of stimulation was negatively correlated with the degree of branching of the added dextran. The results refute the concept that growth of water-insoluble glucan occurs from the multiple, non-reducing termini of dextran acceptors. MH - Carbon Radioisotopes ; Dextrans/*METABOLISM ; Glucans/BIOSYNTHESIS ; Glucosyltransferases/ISOLATION & PURIFICATION/*METABOLISM ; Kinetics ; Streptococcus mutans/*ENZYMOLOGY ; Substrate Specificity ; Sucrose/ METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Carbohydr Res 1986 Feb 1;146(2):259-70 19 UI - 86157542 AU - Steele JL ; McKay LL TI - Partial characterization of the genetic basis for sucrose metabolism and nisin production in Streptococcus lactis. AB - We attempted to identify the genetic loci for sucrose-fermenting ability (Suc+), nisin-producing ability (Nip+), and nisin resistance (Nisr) in certain strains of Streptococcus lactis. To obtain genetic evidence linking the Suc+ Nip+ Nisr phenotype to a distinct plasmid, both conjugal transfer and transformation were attempted. A conjugation procedure modified to protect the recipients against the inhibitory action of nisin allowed the conjugal transfer of the Suc+ Nip+ Nisr marker from three Suc+ Nip+ Nisr donors to various recipients. The frequency of transfer ranged from 1.7 x 10(-4) to 5.6 x 10(-8) per input donor, depending on the mating pair. However, no additional plasmid DNA was apparent in these transconjugants. Transformation of S. lactis LM0230 to the Suc+ Nip+ Nisr phenotype by using the plasmid pool of S. lactis ATCC 11454 was not achieved, even though other plasmids present in the pool were successfully transferred. However, two results imply the involvement of plasmid DNA in coding for the Suc+ Nip+ Nisr phenotype. The Suc+ Nip+ Nisr marker was capable of conjugal transfer to a recipient deficient in host-mediated homologous recombination (Rec-), and the Suc+ Nip+ Nisr marker exhibited bilateral plasmid incompatibility with a number of lactose plasmids found in S. lactis. Although our results indicate that the Suc+ Nip+ Nisr phenotype is plasmid encoded, no physical evidence linking this phenotype to a distinct plasmid was obtained. MH - Conjugation, Genetic ; DNA, Bacterial/ANALYSIS ; Nisin/*BIOSYNTHESIS ; Phenotype ; Plasmids ; Streptococcus Lactis/GENETICS/*METABOLISM ; Sucrose/*METABOLISM ; Transformation, Bacterial SO - Appl Environ Microbiol 1986 Jan;51(1):57-64 20 UI - 86110570 AU - Nealon TJ ; Beachey EH ; Courtney HS ; Simpson WA TI - Release of fibronectin-lipoteichoic acid complexes from group A streptococci with penicillin. AB - Fibronectin binds to Streptococcus pyogenes, and this binding is inhibited by lipoteichoic acid (LTA). Previous studies have shown that LTA can be released from S. pyogenes by treatment with penicillin. Penicillin released LTA from both S. pyogenes and Staphylococcus aureus; however, the binding of fibronectin correlated with the amount of LTA released only in the case of S. pyogenes. Contrarily, clindamycin decreased the ability of S. aureus to bind fibronectin without affecting the binding of fibronectin to S. pyogenes. Further studies indicated that LTA does not inhibit the binding of fibronectin to S. aureus. Fibronectin bound to S. pyogenes could be released from the cell surface by penicillin. Immunological analysis of the released fibronectin indicated that LTA was the only surface component which could be detected as a soluble complex with the released fibronectin. These studies suggest that LTA plays a central role in the binding of fibronectin to S. pyogenes and is not involved in the binding of fibronectin to S. aureus. MH - Adhesiveness ; Animal ; Clindamycin/PHARMACODYNAMICS ; Fibronectins/ ADMINISTRATION & DOSAGE/*METABOLISM ; Penicillins/*PHARMACODYNAMICS ; Phosphatidic Acids/ADMINISTRATION & DOSAGE/IMMUNOLOGY/*METABOLISM/ PHARMACODYNAMICS ; Rabbits ; Staphylococcus aureus/METABOLISM ; Streptococcus Pyogenes/DRUG EFFECTS/*METABOLISM ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; Teichoic Acids/ADMINISTRATION & DOSAGE/IMMUNOLOGY/*METABOLISM/PHARMACODYNAMICS SO - Infect Immun 1986 Feb;51(2):529-35 21 UI - 86094374 AU - Ambudkar SV ; Sonna LA ; Maloney PC TI - Variable stoichiometry of phosphate-linked anion exchange in Streptococcus lactis: implications for the mechanism of sugar phosphate transport by bacteria. AB - Phosphate/2-deoxyglucose 6-phosphate antiport in Streptococcus lactis showed an exchange stoichiometry that varied over a 2-fold range when assay pH was shifted between pH 8.2 and pH 5.2. At pH 7.0 and above, 2 mol of phosphate moved per mol of sugar phosphate; at pH 6.1 the ratio was 1.5:1, while at pH 5.2 the overall stoichiometry fell to 1.1:1. This pattern was not affected by valinomycin in potassium-based media, nor could variable stoichiometry be attributed to altered hydrolysis of the sugar phosphate substrate. In kinetic studies at pH 7.0 or pH 5.2, sugar 6-phosphate was a competitive inhibitor of phosphate transport, indicating operation of a single system. Parallel tests showed that the affinity of antiport for its sugar 6-phosphate substrate was insensitive to pH in this range. Overall, such results suggest a neutral exchange that has specificity for monovalent phosphate but that selects randomly among the available mono- and divalent sugar 6-phosphates. A simple model that shows this behavior suggests a mechanistic role for anion exchange in bacterial transport of sugar phosphate or other organic anions. MH - Anions/*METABOLISM ; Biological Transport ; Hexosephosphates/*METABOLISM ; Hydrogen-Ion Concentration ; Kinetics ; Phosphatases/METABOLISM ; Phosphates/*METABOLISM ; Streptococcus Lactis/*METABOLISM ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Jan;83(2):280-4 22 UI - 86084422 AU - Germaine GR ; Tellefson LM TI - Effect of endogenous phosphoenolpyruvate potential on fluoride inhibition of glucose uptake by Streptococcus mutans. AB - The fluoride sensitivity of glucose uptake by whole cell suspensions of Streptococcus mutans was studied. Preincubation of the organism with up to 1 mM glucose markedly reduced the fluoride sensitivity of subsequent glucose uptake at pH 7.0 and 5.5. Glucose preincubation was shown to result in the establishment of a stable pool of three-carbon glycolytic intermediates. On the basis of inhibition studies and thin-layer chromatography of cell extracts, we suggest that 3- and 2-phosphoglycerate are the principal constituents of the pool. Increased concentrations of glucose used in preincubation mixtures was associated with increased pool sizes of the glycolytic intermediates and increased fluoride resistance. Transport of 2-deoxy-D-glucose by permeabilized cells was inhibited by fluoride when 2-phoshoglycerate served as the energy source. Increased concentrations of 2-phosphoglycerate were shown to overcome the fluoride inhibition of transport. The data suggest that establishment of a stable pool of glycolytic intermediates that includes 2-phosphoglycerate (or its progenitors) may contribute significantly to the apparent refractoriness of plaque microbes to fluoride in vivo. MH - Drug Resistance, Microbial ; Fluorides/*PHARMACODYNAMICS ; Glucose/ *METABOLISM ; Glycerophosphates/METABOLISM ; Phosphoenolpyruvate/ *METABOLISM ; Phosphoenolpyruvate Sugar Phosphotransferase System/ ANTAGONISTS & INHIBITORS ; Phosphopyruvate Hydratase/ANTAGONISTS & INHIBITORS/METABOLISM ; Saliva/*MICROBIOLOGY/PHYSIOLOGY ; Streptococcus mutans/DRUG EFFECTS/*PHYSIOLOGY ; Support, U.S. Gov't, P.H.S. SO - Infect Immun 1986 Jan;51(1):119-24