Polyphosphate Formation Research Articles

Control of vertebrate skeletal mineralization by polyphosphates.

BackgroundSkeletons are formed in a wide variety of shapes, sizes, and compositions of organic and mineral components. Many invertebrate skeletons are constructed from carbonate or silicate minerals, whereas vertebrate skeletons are instead composed of a calcium phosphate mineral known as apatite. No one yet knows why the dynamic vertebrate skeleton, which is continually rebuilt, repaired, and resorbed during growth and normal remodeling, is composed of apatite. Nor is the control of bone and calcifying cartilage mineralization well understood, though it is thought to be associated with phosphate-cleaving proteins. Researchers have assumed that skeletal mineralization is also associated with non-crystalline, calcium- and phosphate-containing electron-dense granules that have been detected in vertebrate skeletal tissue prepared under non-aqueous conditions. Again, however, the role of these granules remains poorly understood. Here, we review bone and growth plate mineralization before showing that polymers of phosphate ions (polyphosphates: (PO3 −)n) are co-located with mineralizing cartilage and resorbing bone. We propose that the electron-dense granules contain polyphosphates, and explain how these polyphosphates may play an important role in apatite biomineralization.Principal Findings/MethodologyThe enzymatic formation (condensation) and destruction (hydrolytic degradation) of polyphosphates offers a simple mechanism for enzymatic control of phosphate accumulation and the relative saturation of apatite. Under circumstances in which apatite mineral formation is undesirable, such as within cartilage tissue or during bone resorption, the production of polyphosphates reduces the free orthophosphate (PO4 3−) concentration while permitting the accumulation of a high total PO4 3− concentration. Sequestering calcium into amorphous calcium polyphosphate complexes can reduce the concentration of free calcium. The resulting reduction of both free PO4 3− and free calcium lowers the relative apatite saturation, preventing formation of apatite crystals. Identified in situ within resorbing bone and mineralizing cartilage by the fluorescent reporter DAPI (4′,6-diamidino-2-phenylindole), polyphosphate formation prevents apatite crystal precipitation while accumulating high local concentrations of total calcium and phosphate. When mineralization is required, tissue non-specific alkaline phosphatase, an enzyme associated with skeletal and cartilage mineralization, cleaves orthophosphates from polyphosphates. The hydrolytic degradation of polyphosphates in the calcium-polyphosphate complex increases orthophosphate and calcium concentrations and thereby favors apatite mineral formation. The correlation of alkaline phosphatase with this process may be explained by the destruction of polyphosphates in calcifying cartilage and areas of bone formation.Conclusions/SignificanceWe hypothesize that polyphosphate formation and hydrolytic degradation constitute a simple mechanism for phosphate accumulation and enzymatic control of biological apatite saturation. This enzymatic control of calcified tissue mineralization may have permitted the development of a phosphate-based, mineralized endoskeleton that can be continually remodeled.

Phosphate Is Essential for Inhibition of the Mitochondrial Permeability Transition Pore by Cyclosporin A and by Cyclophilin D Ablation

Energized mouse liver mitochondria displayed the same calcium retention capacity (a sensitive measure of the propensity of the permeability transition pore (PTP) to open) irrespective of whether phosphate, arsenate, or vanadate was the permeating anion. Unexpectedly, however, phosphate was specifically required for PTP desensitization by cyclosporin A (CsA) or by genetic inactivation of cyclophilin D (CyP-D). Indeed, when phosphate was replaced by arsenate, vanadate, or bicarbonate, the inhibitory effects of CsA and of CyP-D ablation on the PTP disappeared. After loading with the same amount of Ca(2+) in the presence of arsenate or vanadate but in the absence of phosphate, the sensitivity of the PTP to a variety of inducers was identical in mitochondria from wild-type mice, CyP-D-null mice, and wild-type mice treated with CsA. These findings call for a reassessment of conclusions on the role of the PTP in cell death that are based on the effects of CsA or of CyP-D ablation.

Active surface formation on a promoted copper catalyst of the partial oxidation of ethanol

The catalytic properties, chemical composition, and morphology of the surface of electrolytic copper crystals promoted with phosphorus as catalysts of the partial oxidation of ethanol into acetaldehyde were studied. A mechanism of active copper surface formation under the action of the promoting admixture was suggested. Promotion was shown to cause the formation of copper polyphosphates on the surface. Particles of copper metal responsible for selective alcohol transformation are formed in a promoter layer under the action of the reaction medium.

Effect of acrylic polymer and nanocomposite with nano-SiO 2 on thermal degradation and fire resistance of APP–DPER–MEL coating

Acrylic nanocomposite and flame retardant coatings with different acrylic polymers were prepared. The effect of molecular structure and molecular weight of acrylic resins and nanocomposite with nano-SiO 2 on the interaction and char formation of ammonium polyphosphate–dipentaerythritol–melamine (APP–DPER–MEL) coating was investigated using differential thermal analysis (DTA), thermogravimetry (TG), Limiting Oxygen Index (LOI), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and fire protection test. The interaction of APP, DPER, MEL and 3F-1 acrylic resin led to the formation of intumescent coherent char at 300–450 °C. Owing to low molecular weight and lack of benzene rings, F-963 acrylic resin decomposed at lower temperature than APP, and hence their endothermic interaction was destroyed. The well-distributed nano-SiO 2 particles in acrylic nanocomposite could modify char formation and anti-oxidation of char structure at high temperature. It is noted that the fire protection properties of nanocoating with acrylic nanocomposite were better than those of flame retardant coatings with conventional acrylic resins.

Effect of reduced pH on inorganic polyphosphate accumulation by Burkholderia cepacia complex isolates

Burkholderia cepacia complex (Bcc) isolates causing pulmonary infection in cystic fibrosis (CF) patients grow within an acidic environment in the lung. As exposure to acid pH has been shown to increase intracellular inorganic polyphosphate (polyP) formation in some bacteria, we investigated the inter-relationship between acidic pH and polyP accumulation in Bcc isolates. The formation of polyP by one Burkholderia cenocepacia clinical isolate was initially examined at a range of pH values by measuring total intracellular polyP accumulation and phosphate uptake. The pattern of polyP accumulation corresponded with the pattern of phosphate uptake with the maximum for both occurring at pH 5.5. Phosphate uptake and formation of polyP by this isolate was further determined over 48 h at pH 5.5, 6.5 and 7.5; formation of polyP was maximal at pH 5.5 at all time points studied. Sixteen of 17 additional clinical and environmental Bcc isolates examined also exhibited maximum phosphate uptake at pH 5.5. Both clinical and environmental Bcc isolates, of five genomovars, show enhanced formation of polyP in an acidic environment. Given both the speculated role of polyP in pathogenesis, cell signalling and biofilm formation and the acidic nature of the CF lung, this may be of considerable clinical importance. Growth of Bcc in an acidic environment, such as that found in the lungs of CF patients may be influenced in part by polyP accumulation.

31P MAS NMR and Raman study of a Co(Zn)MoP/ γ-Al 2O 3 HDS catalyst precursor containing triethylene glycol

The effect of triethylene glycol on phosphate-doped Co(Zn)Mo catalysts supported on γ-Al 2O 3 was studied by means of 31P magic angle spinning nuclear magnetic resonance (MAS NMR) and Raman spectroscopy. 31P MAS NMR experiments, carried out on ZnMo/ γ-Al 2O 3 doped with phosphate and triethylene glycol, suggested that the P 2 Mo 5 O 23 6 − species that formed in the impregnation solution decomposed to monophosphate and an Anderson-type aluminum molybdate species on contact with the support. The presence of triethylene glycol favored the formation of polyphosphate and PMo 12 O 40 3 − heteropolymolybdate species, but no P 2 Mo 5 O 23 6 − was detected. The use of paramagnetic cobalt instead of zinc cations had a paramagnetic effect on the 31P-NMR signal of the PMo 12 O 40 3 − , indicating that the PMo 12 O 40 3 − species are bound to the promoter atoms. The close proximity of the promoter and Mo allows a better decoration of MoS 2 and improves the catalytic activity.

Mechanism of the formation and properties of antimony polyphosphate

Formation of antimony polyphosphate using Sb2O3 and/or (NH4)2HPO4 and NH4H2PO4 as starting materials has been simulated by thermal analysis technique. The elimination of water and ammonia molecules induced by heating leads to the formation of intermediate ammonium polyphosphate, which subsequently reacts with Sb2O3. Morphologically, vitreous Sb(PO3)3 is composed of plaques having irregular shapes. Infrared spectra and NMR study is consistent with tetrametaphosphate anion arrangement. The compound is thermally unstable and may be recommended as a donor of -O-P-O- linkers in the preparation of special phosphate glasses.

Malikia granosa gen. nov., sp. nov., a novel polyhydroxyalkanoate- and polyphosphate-accumulating bacterium isolated from activated sludge, and reclassification of Pseudomonas spinosa as Malikia spinosa comb. nov.

A Gram-negative, motile, rod-shaped bacterium, designated strain P1(T), was isolated from activated sludge of a municipal wastewater treatment plant. Phylogenetic analysis of its 16S rRNA gene sequence placed the novel isolate among representatives of the family Comamonadaceae. The closest relatives in reconstructed phylogenetic trees were Pseudomonas spinosa, Macromonas bipunctata and Hydrogenophaga species. Strain P1(T) was not able to grow anaerobically or autotrophically, reduced nitrate to nitrite and required vitamins for growth. Ubiquinone 8 (Q8) and 3-hydroxy-substituted fatty acids were present, but 2-hydroxy fatty acids were absent. The G+C content of the DNA was 67 mol%. Phenotypic characteristics allowed a clear differentiation of strain P1(T) from representatives of the genera Hydrogenophaga and Macromonas, whereas DNA-DNA hybridization experiments revealed that strain P1(T) did not belong to the species P. spinosa. As a peculiarity, cells of strain P1(T) and P. spinosa ATCC 14606(T) were able to accumulate large amounts of polyhydroxyalkanoates and polyphosphate in the form of large intracellular granules. Apparently in both strains nitrogen limitation stimulates the production of polyhydroxyalkanoates, whereas carbon starvation induces the formation of polyphosphates. Based upon phylogenetic and phenotypic evidence, it is proposed to establish the novel taxon Malikia granosa gen. nov., sp. nov., represented by the type strain P1(T) (=DSM 15619(T)=JCM 12706(T)=CIP 108194(T)). The most closely related species of strain P1(T) was P. spinosa. This species has been misclassified, and it is proposed to transfer it to the new genus Malikia as Malikia spinosa gen. nov., comb. nov. The type strain is ATCC 14606(T) (=DSM 15801(T)).

THERMAL REACTIONS IN SYNTHETIC APATITE–AMMONIUM SULFATE MIXTURE

The thermal reactions in the mixtures of hydroxylapatite or fluorapatite and (NH4)2SO4up to 500°C were studied with the purpose of elaborating the conditions of obtaining calcium–ammonium cyclophosphate that could be used as fertilizer. Thermal analysis with a simultaneous FTIR analysis of the evolved gases as well as the analyses of chemical and phase composition of solid products were performed. The thermal changes in the mixtures could be divided into three steps: (1) decomposition of (NH4)2SO4and reactions of apatite with these products at 250–420°C, (2) calcium ammonium polyphosphate formation at 290–450°C, and (3) reaction of CaSO4with CaNH4P3O9at 320–500°C. Higher concentrations of NH3in the gas phase promote the formation of CaNH4P3O9and increase its stability. Calcination at temperatures above 350°C causes decomposition of CaNH4P3O9with a decrease in the content of water-soluble phosphorus and evolvement of SO2.

Preparation and characterization of sol–gel derived glasses in the ternary Na2O–Al2O3–P2O5system

A new sol–gel synthesis route to organic free Na2O–Al2O3–P2O5 xerogels was developed. These xerogels contain P2O5 up to 67.5 mole % and Al2O3 up to 20 mole % and could be melted to homogeneous glasses in air at significantly lower temperatures than those applied using the conventionally melting method. Aluminum isopropoxide dissolved in isopropanol, orthophosphoric acid dissolved in isopropanol and sodium nitrate were used as precursors for Al2O3, P2O5 and Na2O respectively. Clear stable alcoholic solutions were prepared in the presence of HNO3. Addition of HNO3 in the concentration used, enabled homogeneous mixing of the precursors, controlled over the gelation step and prevented the fast precipitation of the AlPO4 crystalline phase which breaks down the glass formation ability. The sol–gel route developed was investigated using different analytical tools. Differential thermal analysis (DTA) coupled with thermo-gravimetric (TG) analysis indicated that neither organic residues nor sodium nitrate residues were present in the dried gels after gelation. Fourier transform infrared spectroscopy (FTIR) showed the presence of all characteristic phosphate groups and bonds in the dried gels, especially the P=O and the P–O–P bonds. Solid state 31P Magic angle spinning nuclear magnetic resonance (MAS NMR) revealed the formation of Q2 chain-like polyphosphates of variable polymerization degrees in the dried gels. Solid state 27Al MAS NMR revealed that octahedrally coordinated Al is the preferred moiety in the dried gels. However, powder X-ray diffraction (XRD) investigation indicated the presence of crystalline Q1 oligomeric pyrophosphate species in the dried gels. Isotropic chemical shifts belonging to these pyrophosphate species were also recorded in the 31P MAS NMR spectra. Characterization of the prepared glasses showed that their properties are in very good agreement with those glasses prepared by the conventionally melting method.

Biological uptake of Cd, Se(IV) and Zn by Chlamydomonas reinhardtii in response to different phosphate and nitrate additions

We investigated the influences of different nutrient regimes on the accumulation of Cd, Zn, and Se(IV) in the freshwater green alga Chlamydomonas reinhardtii over a 4 h exposure period. After the cells had been acclimated to different ambient P levels from 0.1 to 10.0 µM for 2 d in differ- ent media (buffer, basic and simplified uptake media), their intracellular metal concentrations increased by 3.6 to 14× for Cd and 1.6 to 4.0× for Zn. Se uptake was, however, decreased by 7.7 to 44× . Semi-continuous culture experiments further demonstrated that the alga's metal concentrations were enhanced 26× for Cd and 8.0× for Zn, whereas Se accumulation was inhibited 75× at the same medium P levels. The uptake rates in semi-continuous cultures increased by 269× for Cd and 11× for Zn, but Se uptake decreased 92× with increasing P concentration. Medium P additions also signifi- cantly increased intracellular partitioning for Cd and Zn, but there was no apparent effect for Se. For P-starved or P-repleted cells, the P-enriched medium dramatically increased the cellular accumula- tion of Cd and Zn compared with the P-depleted medium. A surge uptake of Cd and Zn occurred in P-starved cells. P starvation also resulted in a remarkable increase in Se accumulation in the P- deplete medium. N enrichment from 5 to 200 µM significantly increased the alga's uptake and intra- cellular partitioning for Cd and Zn, although the effects were not strictly concentration-dependent. Our study strongly suggests that ambient P enrichment greatly stimulates Cd and Zn accumulation and sequestration in C. reinhardtii, presumably due to rapid P uptake and subsequent formation of polyphosphate in the cells, and distinctly inhibits Se uptake possibly due to the competition of similar anionic forms for transport between Se and P.

Surface analytical studies of surface-additive interactions, by means of in situ and combinatorial approaches

The influence of tribological conditions on the surface reactions occurring between zinc dialkyldithiophosphate (ZnDTP) and steel surfaces has been studied by means of a combination of X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectroscopy (ToF-SIMS), in situ attenuated total reflection (ATR) infrared spectroscopy, and high-throughput combinatorial approaches. Purely thermal treatment at 150 °C appears to lead to the formation of zinc polyphosphates. However, in the presence of tribological stress, simple phosphates appear to dominate, with some indication that higher load conditions lead to an increase in the surface concentration of both phosphate and, at higher temperatures, polyphosphate.

Study of the Interaction of ZDDP and Dispersants Using X-ray Absorption Near Edge Structure Spectroscopy—Part 1: Thermal Chemical Reactions

The antiwear properties of zinc dialkyldithiophosphate (ZDDP), dispersants, and mixtures of ZDDP and different dispersants have been evaluated using a pin-on-flat Plint wear machine. Tribochemical interactions between ZDDP and dispersants have been investigated under boundary lubrication conditions by means of X-ray absorption near edge structure (XANES) spectroscopy, probing the phosphorus, sulfur and nitrogen absorption edges. The results show that the dispersants do not give any wear protection by themselves in the base oil. The dispersants also do not affect the antiwear property of ZDDP under the given testing conditions. The N K-edge XANES analysis indicates that dispersants contribute to the chemical composition of the tribofilms and form mixed ammonium/zinc polyphosphates. Phosphorus in the tribofilms is present mainly in the form of medium-chain polyphosphate on the surface and short-chain polyphosphate in the bulk. Sulfur appears in the tribofilms mainly as sulfide S-II, possibly zinc sulfide. The presence of dispersants in oil blends does not disturb the polyphosphate (and sulfide) formation, but it does decrease the chain length of the polyphosphate in the tribofilms.

Influence of phosphate modification on the surface properties of sulfated titania

The effect of increasing phosphate loading on the bulk and surface properties of sulfated titania was investigated by thermogravimetry, X-ray diffraction, laser Raman spectroscopy and FT-IR spectroscopy. The acid properties of the material were probed by low-temperature FT-IR spectroscopy of adsorbed CO. Monophosphate species in varying symmetries and protonation states were detected on the surface of the sulfated titania at low loadings. These condensed with formation of surface polyphosphates with increasing loading. Phosphate blocks weak Lewis acid sites and partially displaces sulfate groups. While the acid, properties of phosphate-free sulfated titania are dominated by Lewis acid sites, the phosphated samples also developed Bronsted acid sites due to the presence of POH groups on the surface.

Combined in situ (ATR FT-IR) and ex situ (XPS) Study of the ZnDTP-Iron Surface Interaction

Attenuated total reflection infrared (ATR FT-IR) and X-ray photoelectron spectroscopy (XPS) have been used for the in situ and ex situ characterization of thermal and tribological films formed on iron from a commercial zinc dialkyldithiophosphate (ZnDTP). From in situ ATR FT-IR analysis, information on the chemical changes occurring at the iron/lubricant additive interface was obtained during heating and sliding at high temperatures. Different mechanisms and chemical compositions have been found for the thermal and tribochemical reactions between the ZnDTP and the iron surface under the experimental conditions used in this work. Both the ATR FT-IR and the XPS results show the decomposition of ZnDTP with the formation of polyphosphates following thermal testing at 150 °C. However, after tribological testing at the same temperature an inorganic phosphate film has been detected on the iron surface instead.

Thermodynamic Aspects of the Formation of Potassium Polyphosphate and Polycondensation with Glycerin Dichlorohydrin

Thermodynamic Aspects of the Formation of Potassium Polyphosphate and Polycondensation with Glycerin Dichlorohydrin

Subcellular compartmentation of elements in non‐mycorrhizal and mycorrhizal roots of Pinus sylvestris: an X‐ray microanalytical study. I. The distribution of phosphate

The inter‐ and intracellular distribution of phosphate in non‐mycorrhizal and ectomycorrhizal roots of Pinus sylvestris was analysed by energy‐dispersive X‐ray spectroscopy after cryofixation, freeze‐drying, pressure infiltration with resin and dry cutting of the material. The results show (1) that in non‐mycorrhizal roots the distribution of phosphate across the root cross section is relatively homogenous, (2) that the intracellular distribution of phosphate depends on the external supply, and (3) that the phosphate absorption can be increased by a supply of NH4. Under phosphate starvation, the phosphate is mainly localized in the cell cytoplasm, whereas under high external phosphate this element accumulates in the vacuoles of cortical cells. It can be inferred that at low external conditions phosphate is mainly located in the cytoplasm of cells to protect metabolism from deficiency, that phosphate is translocated to the shoot via symplastic and apoplastic pathways, and that especially the vacuolar pool size is regulated by the external phosphate supply. The nutrition of the host plant under phosphate starvation was improved by a mycorrhizal infection with Suillus bovinus. However, this effect might not be the same for all ectomycorrhizal infections and supply conditions. With high external phosphate an ectomycorrhizal infection with S. bovinus had no effect on intracellular phosphate within the roots, the shoot contents of mycorrhizal plants were less than those of non‐mycorrhizal seedlings and translocation rate across the mycorrhizal interface appeared to be independent of the availability of phosphate. Intracellular phosphate in fungal cells of mycorrhizal roots was independent of the external supply and seemed to be regulated by the formation of polyphosphates. A supply of (NH4)2HPO4, which led to an increase in cytoplasmic phosphate levels in the hyphae of the Hartig net of the Paxillus involutus mycorrhiza also led to higher phosphate in the plant cell compartments. Therefore, it can be inferred that the translocation of phosphate from fungus to host plant across the mycorrhizal interface is regulated by the phosphate concentration in the cytoplasm of the Hartig net and by the efflux rate into the interfacial apoplast.

PauA: a novel plasminogen activator fromStreptococcus uberis

Chromosomal DNA from two geographically distinct isolates of Streptococcus uberis was used to clone the plasminogen activator in an active form in Escherichia coli. The cloned fragments from each strain contained four potential open reading frames (ORFs). That for the plasminogen activator encoded a protein of 286 amino acids (33.4 kDa) which is cleaved between residues 25 and 26 during secretion by S. uberis. The amino acid sequence of the mature protein showed only weak homology (23.5-28%) to streptokinase. The plasminogen activator gene, pauA, in S. uberis was located between two ORFs with high homology to the DNA mismatch repair genes, hexA and hexB, and not on a DNA fragment between the genes encoding an ATP binding cassette transporter protein (abc) and a protein involved in the formation and degradation of guanosine polyphosphates (rel) as is the case for streptokinase in other streptococci.

PauA: a novel plasminogen activator from Streptococcus uberis

Chromosomal DNA from two geographically distinct isolates of Streptococcus uberis was used to clone the plasminogen activator in an active form in Escherichia coli. The cloned fragments from each strain contained four potential open reading frames (ORFs). That for the plasminogen activator encoded a protein of 286 amino acids (33.4 kDa) which is cleaved between residues 25 and 26 during secretion by S. uberis. The amino acid sequence of the mature protein showed only weak homology (23.5–28%) to streptokinase. The plasminogen activator gene, pauA, in S. uberis was located between two ORFs with high homology to the DNA mismatch repair genes, hexA and hexB, and not on a DNA fragment between the genes encoding an ATP binding cassette transporter protein ( abc) and a protein involved in the formation and degradation of guanosine polyphosphates ( rel) as is the case for streptokinase in other streptococci.

Polyphosphate formation by Acinetobacter johnsonii 210A: effect of cellular energy status and phosphate-specific transport system

In acetate-limited chemostat cultures of Acinetobacter johnsonii 210A at a dilution rate of 0.1 h−1 the polyphosphate content of the cells increased from 13% to 24% of the biomass dry weight by glucose (100 mM), which was only oxidized to gluconic acid. At this dilution rate, only about 17% of the energy from glucose oxidation was calculated to be used for polyphosphate synthesis, the remaining 83% being used for biomass formation. Suspensions of non-growing, phosphate-deficient cells had a six- to tenfold increased uptake rate of phosphate and accumulated polyphosphate aerobically up to 53% of the biomass dry weight when supplied with only orthophosphate and Mg2+. The initial polyphosphate synthesis rate was 98 ± 17 nmol phosphate min−1 mg protein−1. Intracellular poly-β-hydroxybutyrate and lipids served as energy sources for the active uptake of phosphate and its subsequent sequestration to polyphosphate. The H+-ATPase inhibitor N,N′-dicyclohexylcarbodiimide caused low ATP levels and a severe inhibition of polyphosphate formation, suggesting the involvement of polyphosphate kinase in polyphosphate synthesis. It is concluded that, in A. johnsonii 210A, (i) polyphosphate is accumulated as the energy supply is in excess of that required for biosynthesis, (ii) not only intracellular poly-β-hydroxybutyrate but also neutral lipids can serve as an energy source for polyphosphate-kinase-mediated polyphosphate formation, (iii) phosphate-deficient cells may accumulate as much polyphosphate as activated sludges and recombinants of Escherichia coli designed for polyphosphate accumulation.