scholarly journals Teichoic acid synthesis in Bacillus stearothermophilus

1974 ◽  
Vol 138 (3) ◽  
pp. 525-535 ◽  
Author(s):  
L. D. Kennedy
Keyword(s):  
Alkaline Hydrolysis ◽  
Bacillus Stearothermophilus ◽  
De Novo ◽  
Enzyme System ◽  
Teichoic Acid ◽  
Periodate Oxidation ◽  
Acid Synthesis ◽  
Pulse Labelling ◽  
Glycerol Phosphate ◽  
Enzyme Preparations

1. Particulate enzyme preparations obtained from Bacillus stearothermophilus B65 by digestion with lysozyme were shown to catalyse teichoic acid synthesis. With CDP-glycerol as sole substrate the preparations synthesized 1,3-poly(glycerol phosphate). It was characterized by alkaline hydrolysis, by glucosylation to the alkali-stable 2-glucosyl-1,3-poly(glycerol phosphate) with excess of UDP-glucose and a Bacillus subtilis Marburg enzyme system, by degradation of this latter product with 60%HF and periodate oxidation of the resulting glucosylglycerol. The specificity of the B. subtilis system previously reported (Glaser & Burger, 1964), was confirmed in the present work. 2. Pulse-labelling experiments, followed by periodate oxidation of the product and isolation of formaldehyde from the glycerol terminus of the polymer, showed that the B. stearothermophilus enzyme system transferred glycerol phosphate units to the glycerol end of the chain. The transfer reaction was irreversible. It was not determined if these poly(glycerol phosphate) chains were synthesized de novo, but it was shown that the newly synthesized oligomers were bound to much larger molecules. 3. When the B. stearothermophilus enzyme system was supplied with both CDP-glycerol and UDP-glucose, 1-glucosyl-2,3-poly(glycerol phosphate) was synthesized in addition to the 1,3-isomer. The former polymer was characterized by acid and alkaline hydrolysis, degradation with HF and periodate oxidation of the resulting glucosylglycerol, and periodate oxidation of the intact polymer followed by mild acid hydrolysis. This latter procedure removed the glucose substituents without disrupting the poly(glycerol phosphate) chain. 4. The poly(glycerol phosphate) isomers were distinguished by glucosylation with the B. subtilis enzymes and alkaline hydrolysis, the 2,3-isomer remaining alkali-labile. The proportion of 2,3-poly(glycerol phosphate) in the product increased with increasing amounts of UDP-glucose in the incubation mixture, but the total glycerol phosphate incorporated into products remained constant. It is suggested that the synthetic pathways of the two poly(glycerol phosphate) species may share a rate-limiting step.

scholarly journals The biosynthesis of the wall teichoic acid in Staphylococcus lactis I3

1968 ◽  
Vol 110 (3) ◽  
pp. 565-571 ◽  
Author(s):  
J Baddiley ◽  
N. L. Blumsom ◽  
L. Julia Douglas
Keyword(s):  
Enzyme System ◽  
Teichoic Acid ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Enzyme Preparations

1. The biosynthesis of the wall teichoic acid in Staphylococcus lactis I3 was studied. Cell-free particulate enzyme preparations, probably representing fragmented membrane, were isolated and used for the synthesis of polymer. 2. By using appropriately labelled CDP-glycerol and UDP-N-acetylglucosamine it was shown that the former contributes a glycerol phosphate residue and the latter contributes an N-acetylglucosamine 1-phosphate residue to the repeating unit. 3. No polymer was synthesized unless both nucleotides were present, and no other substrates were required. 4. The properties of the enzyme system were studied. 5. Although attempts to fractionate the system failed, the biosynthesis is believed to be complex and its mechanism is considered.

scholarly journals Biosynthesis of the wall teichoic acid in Bacillus licheniformis

1972 ◽  
Vol 127 (1) ◽  
pp. 27-37 ◽  
Author(s):  
I. C. Hancock ◽  
J. Baddiley
Keyword(s):  
Bacillus Licheniformis ◽  
Teichoic Acid ◽  
Chain Extension ◽  
Pulse Labelling ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Anomeric Configuration ◽  
Glucose Phosphate ◽  
Chemical Studies ◽  
Two Stages

1. The biosynthesis of the wall teichoic acid, poly(glycerol phosphate glucose), has been studied with a particulate membrane preparation from Bacillus licheniformis A.T.C.C. 9945. The precursor CDP-glycerol supplies glycerol phosphate residues, whereas UDP-glucose supplies only glucose to the repeating structure of the polymer. 2. Synthesis proceeds through polyprenol phosphate derivatives, and chemical studies and pulse-labelling techniques show that the first intermediate is the phosphodiester, glucose polyprenol monophosphate. CDP-glycerol donates a glycerol phosphate residue to this to give a second intermediate, (glycerol phosphate glucose phosphate) polyprenol. 3. The glucose residue in the lipid intermediates has the β configuration, and chain extension in the synthesis of polymer occurs by transglycosylation with inversion of anomeric configuration at two stages.

scholarly journals The mechanism of biosynthesis and direction of chain extension of a polyl-(N-acetylglucosamine 1-phosphate) from the walls of Staphylococcus lactis N.C.T.C. 2102

1969 ◽  
Vol 113 (4) ◽  
pp. 635-642 ◽  
Author(s):  
D. Brooks ◽  
J Baddiley
Keyword(s):  
Enzyme System ◽  
Group Analysis ◽  
Chain Extension ◽  
Pulse Labelling ◽  
Uridine Diphosphate ◽  
High Concentration ◽  
Ph Optimum ◽  
Enzyme Preparations ◽  
End Group

1. The synthesis of a polymer of N-acetylglucosamine 1-phosphate, occurring in the walls of Staphylococcus lactis N.C.T.C. 2102, was examined by using cell-free enzyme preparations. The enzyme system was particulate, and probably represents fragmented cytoplasmic membrane. 2. Uridine diphosphate N-acetylglucosamine was the only substrate required for polymer synthesis and labelled substrate was used to show that N-acetylglucosamine 1-phosphate is transferred as an intact unit from substrate to polymer. 3. The properties of the enzyme system were studied. A high concentration of Mg2+ or Mn2+ was required for optimum activity, and the pH optimum was about 8·5. 4. End-group analysis during synthesis in vitro showed that newly formed chains contain up to about 15 repeating units. Pulse-labelling indicated that chain extension occurs by transfer from the nucleotide to the ‘sugar-end’ of the chain, i.e. to the end that is not attached to peptidoglycan in the wall.

scholarly journals The Amino Terminus of Bacillus subtilis TagB Possesses Separable Localization and Functional Properties

2007 ◽  
Vol 189 (19) ◽  
pp. 6816-6823 ◽  
Author(s):  
Amit P. Bhavsar ◽  
Michael A. D'Elia ◽  
Tiffany D. Sahakian ◽  
Eric D. Brown
Keyword(s):  
Bacillus Subtilis ◽  
Protein Function ◽  
Teichoic Acid ◽  
Acid Synthesis ◽  
Amino Terminus ◽  
Membrane Localization ◽  
Membrane Targeting ◽  
Glycerol Phosphate ◽  
Wall Teichoic Acid ◽  
Necessary And Sufficient

ABSTRACT The function(s) of gram-positive wall teichoic acid is emerging with recent findings that it is an important virulence factor in the pathogen Staphylococcus aureus and that it is crucial to proper rod-shaped cell morphology of Bacillus subtilis. Despite its importance, our understanding of teichoic acid biosynthesis remains incomplete. The TagB protein has been implicated in the priming step of poly(glycerol phosphate) wall teichoic acid synthesis in B. subtilis. Work to date indicates that the TagB protein is localized to the membrane, where it adds a single glycerol phosphate residue to the nonreducing end of the undecaprenol-phosphate-linked N-acetylmannosamine-β(1,4)-N-acetylglucosamine-1-phosphate. Thus, membrane association is critical to TagB function. In this work we elucidate the mechanism of TagB membrane localization. We report the identification of a membrane targeting determinant at the amino terminus of TagB that is necessary and sufficient for membrane localization. The putative amphipathicity of this membrane targeting determinant was characterized and shown to be required for TagB function but not localization. This work shows for the first time that the amino terminus of TagB mediates membrane targeting and protein function.

scholarly journals Poly(glucosylglycerol phosphate) teichoic acid in the walls of Bacillus stearothermophilus B65

1975 ◽  
Vol 151 (1) ◽  
pp. 115-120 ◽  
Author(s):  
A J Anderson ◽  
A R Archibald
Keyword(s):  
Concanavalin A ◽  
Bacillus Stearothermophilus ◽  
Teichoic Acid ◽  
Acid Form ◽  
Glycerol Phosphate ◽  
Phosphodiester Linkage

1. Walls of Bacillus stearothermophilus B65 contain a glycerol teichoic acid in which repeating structures consisting of 1-O-α-D-glucopyranosylglycerol phosphate are held together by phosphodiester linkage between the glycerol and glucose moieties of adjacent units. 2. The walls are not agglutinated on incubation with concanavalin A, nor does the isolated teichoic acid form a precipitate with this lectin. 3. No evidence was obtained of the presence of the glucosylated (1 leads to 2)-poly(glycerol phosphate) teichoic acid which has previously been reported to occur in walls of this bacterium.

scholarly journals Synthesis of fatty acids in the perfused mouse liver

1974 ◽  
Vol 142 (3) ◽  
pp. 611-618 ◽  
Author(s):  
D. Michael W. Salmon ◽  
Neil L. Bowen ◽  
Douglas A. Hems
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
De Novo ◽  
Saturated Fatty Acids ◽  
Carbon Sources ◽  
Acid Synthesis ◽  
Hepatic Glycogen ◽  
Total Rate ◽  
Glucose Carbon

1. Fatty acid synthesis de novo was measured in the perfused liver of fed mice. 2. The total rate, measured by the incorporation into fatty acid of3H from3H2O (1–7μmol of fatty acid/h per g of fresh liver), resembled the rate found in the liver of intact mice. 3. Perfusions with l-[U-14C]lactic acid and [U-14C]glucose showed that circulating glucose at concentrations less than about 17mm was not a major carbon source for newly synthesized fatty acid, whereas lactate (10mm) markedly stimulated fatty acid synthesis, and contributed extensive carbon to lipogenesis. 4. The identification of 50% of the carbon converted into newly synthesized fatty acid lends further credibility to the use of3H2O to measure hepatic fatty acid synthesis. 5. The total rate of fatty acid synthesis, and the contribution of glucose carbon to lipogenesis, were directly proportional to the initial hepatic glycogen concentration. 6. The proportion of total newly synthesized lipid that was released into the perfusion medium was 12–16%. 7. The major products of lipogenesis were saturated fatty acids in triglyceride and phospholipid. 8. The rate of cholesterol synthesis, also measured with3H2O, expressed as acetyl residues consumed, was about one-fourth of the basal rate of fatty acid synthesis. 9. These results are discussed in terms of the carbon sources of hepatic newly synthesized fatty acids, and the effect of glucose, glycogen and lactate in stimulating lipogenesis, independently of their role as precursors.

scholarly journals The linkage of sugar phosphate polymer to peptidoglycan in walls of Micrococcus sp. 2102

1978 ◽  
Vol 169 (2) ◽  
pp. 329-336 ◽  
Author(s):  
J Heptinstall ◽  
J Coley ◽  
P J Ward ◽  
A R Archibald ◽  
J Baddiley
Keyword(s):  
Staphylococcus Aureus ◽  
Alkaline Hydrolysis ◽  
The Other ◽  
Muramic Acid ◽  
Sugar Phosphate ◽  
Glycerol Phosphate ◽  
Homogeneous Product ◽  
A Chain

1. Protein-free walls of Micrococcus sp. 2102 contain peptidoglycan, poly-(N-acetylglucosamine 1-phosphate) and small amounts of glycerol phosphate. 2. After destruction of the poly-(N-acetylglucosamine 1-phosphate) with periodate, the glycerol phosphate remains attached to the wall, but can be removed by controlled alkaline hydrolysis. The homogeneous product comprises a chain of three glycerol phosphates and an additional phosphate residue. 3. The poly-(N-acetylglucosamine 1-phosphate) is attached through its terminal phosphate to one end of the tri(glycerol phosphate). 4. The other end of the glycerol phosphate trimer is attached through its terminal phosphate to the 3-or 4-position of an N-acetylglucosamine. It is concluded that the sequence of residues in the sugar 1-phosphate polymer-peptidoglycan complex is: (N-acetylglucosamine 1-phosphate)24-(glycerol phosphate)3-N-acetylglucosamine 1-phosphate-muramic acid (in peptidoglycan). Thus in this organism the phosphorylated wall polymer is attached to the peptidoglycan of the wall through a linkage unit comprising a chain of three glycerol phosphate residues and an N-acetylglucosamine 1-phosphate, similar to or identical with the linkage unit in Staphylococcus aureus H.

scholarly journals High glucose levels reduce fatty acid oxidation and increase triglyceride accumulation in human placenta

2013 ◽  
Vol 305 (2) ◽  
pp. E205-E212 ◽  
Author(s):  
Francisco Visiedo ◽  
Fernando Bugatto ◽  
Viviana Sánchez ◽  
Irene Cózar-Castellano ◽  
Jose L. Bartha ◽  
...  
Keyword(s):  
Fatty Acid ◽  
High Glucose ◽  
De Novo ◽  
Acid Synthesis ◽  
Acid Oxidation ◽  
Glucose Levels ◽  
Triglyceride Accumulation ◽  
Triglyceride Content ◽  
Low Glucose ◽  
Cpt I

Placentas of women with gestational diabetes mellitus (GDM) exhibit an altered lipid metabolism. The mechanism by which GDM is linked to alterations in placental lipid metabolism remains obscure. We hypothesized that high glucose levels reduce mitochondrial fatty acid oxidation (FAO) and increase triglyceride accumulation in human placenta. To test this hypothesis, we measured FAO, fatty acid esterification, de novo fatty acid synthesis, triglyceride levels, and carnitine palmitoyltransferase activities (CPT) in placental explants of women with GDM or no pregnancy complication. In women with GDM, FAO was reduced by ∼30% without change in mitochondrial content, and triglyceride content was threefold higher than in the control group. Likewise, in placental explants of women with no complications, high glucose levels reduced FAO by ∼20%, and esterification increased linearly with increasing fatty acid concentrations. However, de novo fatty acid synthesis remained unchanged between high and low glucose levels. In addition, high glucose levels increased triglyceride content approximately twofold compared with low glucose levels. Furthermore, etomoxir-mediated inhibition of FAO enhanced esterification capacity by ∼40% and elevated triglyceride content 1.5-fold in placental explants of women, with no complications. Finally, high glucose levels reduced CPT I activity by ∼70% and phosphorylation levels of acetyl-CoA carboxylase by ∼25% in placental explants of women, with no complications. We reveal an unrecognized regulatory mechanism on placental fatty acid metabolism by which high glucose levels reduce mitochondrial FAO through inhibition of CPT I, shifting flux of fatty acids away from oxidation toward the esterification pathway, leading to accumulation of placental triglycerides.

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