Organic Polyphosphates Research Articles

Different profiles of soil phosphorous compounds depending on tree species and availability of soil phosphorus in a tropical rainforest in French Guiana

BackgroundThe availability of soil phosphorus (P) often limits the productivities of wet tropical lowland forests. Little is known, however, about the metabolomic profile of different chemical P compounds with potentially different uses and about the cycling of P and their variability across space under different tree species in highly diverse tropical rainforests.ResultsWe hypothesised that the different strategies of the competing tree species to retranslocate, mineralise, mobilise, and take up P from the soil would promote distinct soil 31P profiles. We tested this hypothesis by performing a metabolomic analysis of the soils in two rainforests in French Guiana using 31P nuclear magnetic resonance (NMR). We analysed 31P NMR chemical shifts in soil solutions of model P compounds, including inorganic phosphates, orthophosphate mono- and diesters, phosphonates, and organic polyphosphates. The identity of the tree species (growing above the soil samples) explained > 53% of the total variance of the 31P NMR metabolomic profiles of the soils, suggesting species-specific ecological niches and/or species-specific interactions with the soil microbiome and soil trophic web structure and functionality determining the use and production of P compounds. Differences at regional and topographic levels also explained some part of the the total variance of the 31P NMR profiles, although less than the influence of the tree species. Multivariate analyses of soil 31P NMR metabolomics data indicated higher soil concentrations of P biomolecules involved in the active use of P (nucleic acids and molecules involved with energy and anabolism) in soils with lower concentrations of total soil P and higher concentrations of P-storing biomolecules in soils with higher concentrations of total P.ConclusionsThe results strongly suggest “niches” of soil P profiles associated with physical gradients, mostly topographic position, and with the specific distribution of species along this gradient, which is associated with species-specific strategies of soil P mineralisation, mobilisation, use, and uptake.

31P-NMR Metabolomics Revealed Species-Specific Use of Phosphorous in Trees of a French Guiana Rainforest.

Productivity of tropical lowland moist forests is often limited by availability and functional allocation of phosphorus (P) that drives competition among tree species and becomes a key factor in determining forestall community diversity. We used non-target 31P-NMR metabolic profiling to study the foliar P-metabolism of trees of a French Guiana rainforest. The objective was to test the hypotheses that P-use is species-specific, and that species diversity relates to species P-use and concentrations of P-containing compounds, including inorganic phosphates, orthophosphate monoesters and diesters, phosphonates and organic polyphosphates. We found that tree species explained the 59% of variance in 31P-NMR metabolite profiling of leaves. A principal component analysis showed that tree species were separated along PC 1 and PC 2 of detected P-containing compounds, which represented a continuum going from high concentrations of metabolites related to non-active P and P-storage, low total P concentrations and high N:P ratios, to high concentrations of P-containing metabolites related to energy and anabolic metabolism, high total P concentrations and low N:P ratios. These results highlight the species-specific use of P and the existence of species-specific P-use niches that are driven by the distinct species-specific position in a continuum in the P-allocation from P-storage compounds to P-containing molecules related to energy and anabolic metabolism.

A Novel Approach for the Quantification of Different Inorganic and Organic Phosphorus Compounds in Environmental Samples by P L2,3-Edge X-ray Absorption Near-Edge Structure (XANES) Spectroscopy.

Phosphorus (P) is an essential element for life on Earth, with an important and oftentimes unaccounted organic biogeochemical component. Current methods for the quantification of different organic P compounds in environmental samples (e.g., soils, sediments) are based on extraction techniques and often associated with incomplete P recovery or sample changes. In this study, we present a protocol for the quantification of different organic and inorganic P species in soils using synchrotron-based X-ray absorption near-edge structure (XANES) spectroscopy at the P L2,3-edge. Its accuracy and precision was evaluated by analyzing 40 standard mixtures composed of seven different inorganic and organic P compounds (with a mean of R2 = 0.85). In addition, we quantified the P species of two soils and two agro-industrial byproducts using P L2,3-edge XANES spectroscopy and the results were compared with those obtained by P K-edge XANES or 31P NMR spectroscopy. Using the P L2,3-edge, we identified different organic Pspecies, including those not identified by the common P K-edge XANES. However, there is a consistent underestimation of organic polyphosphates. Overall, the application of P L2,3-edge XANES provides a higher level of information than by P K-edge XANES, although the ubiquitous use of this novel methodology is still limited to samples with a phosphorus content above 3 mg g-1.

In Situ Investigation of Mammalian Inorganic Polyphosphate Localization Using Novel Selective Fluorescent Probes JC-D7 and JC-D8

Inorganic polyphosphate (polyP) is a polymer composed of many orthophosphates linked together by phosphoanhydride bonds. Recent studies demonstrate that in addition to its important role in the function of microorganisms, polyP plays multiple important roles in the pathological and physiological function of higher eukaryotes, including mammalians. However, due to the dramatically lower abundance of polyP in mammalian cells when comparing to microorganisms, its investigation poses an experimental challenge. Here, we present the identification of novel fluorescent probes that allow for specific labeling of synthetic polyP in vitro as well as endogenous polyP in living cells. These probes demonstrate high selectivity for the labeling of polyP that was not sensitive to a number of ubiquitous organic polyphosphates, notably RNA. Use of these probes allowed us to demonstrate the real time detection of polyP release from lysosomes in live cells. Furthermore, we have been able to detect the increased levels of polyP in cells with Parkinson's disease related mutations.

Phosphorus‐31 Nuclear Magnetic Resonance Spectral Assignments of Phosphorus Compounds in Soil NaOH–EDTA Extracts

Soil P composition can be conveniently determined in alkaline extracts using solution 31P nuclear magnetic resonance (NMR) spectroscopy, but spectral assignments are based on fragmentary literature reports of model compounds in various extraction matrices. We report solution 31P NMR chemical shifts of model P compounds, including inorganic phosphates, orthophosphate monoesters and diesters, phosphonates, and organic polyphosphates, determined in a standardized soil P extractant (0.25 M NaOH and 0.05 M EDTA). Signals from nucleic acids (DNA −0.37 ppm, RNA 0.54 ppm) and phospholipids (phosphatidyl choline 0.78 ppm, phosphatidyl serine 1.57 ppm, phosphatidyl ethanolamine 1.75 ppm) could be differentiated in the orthophosphate diester region, and were identified in a sample of cultured soil bacteria. Inorganic and organic polyphosphates could be differentiated by the presence of a signal at −9 ppm from the α phosphate of organic polyphosphates. Some orthophosphate diesters, notably RNA and phosphatidyl choline, degraded rapidly to orthophosphate monoesters in NaOH–EDTA although DNA, other phospholipids, and orthophosphate monoesters were more stable. Changes in probe temperature had a marked influence on signal intensities and the relative magnitude of signals from orthophosphate monoesters and inorganic orthophosphate, and we suggest that solution 31P NMR spectroscopy of soil extracts be performed at 20°C.

Polyvalent cation effects on myo-inositol hexakis dihydrogenphosphate enzymatic dephosphorylation in dairy wastewater.

Information is needed on organic polyphosphates such as myo-inositol 1,2,3,5/4,6-hexakis dihydrogenphosphate or phytate (IP6) contribution to the sources and sinks of dissolved phosphorus (PO4-P) in the soil-manure-water system. Effects of Na+, Ca2+, Al3+, and Fe3+ and cation to IP6-P mole ratios on the enzymatic dephosphorylation of IP6 were studied to determine controlling mechanisms of dephosphorylation and persistence in manure. Phytate- and PO4-P were analyzed by high-performance liquid chromatography. Phytate dephosphorylation by Aspergillus ficuum (Reichardt) Henn. phytase EC 3.1.3.8 decreases by 50 +/- 3.6 and 40 +/- 4% at pH 4.5 and 6, respectively, as Ca2+ concentrations increase and cation to IP6-P mole ratios reach 6:6. Polyanionic IP6 has a high affinity for Al3+ and Fe3+ and reductions in dephosphorylation average 27 and 32% at a cation to IP6-P mole ratio of 1:6 for Al3+ and Fe3+, respectively, while reaching more than 99% at a mole ratio of 6:6. A phytase-hydrolyzable phosphorus (PHP) fraction is native to ruminant animal manure and is proportional to total solids (TS) concentration in 1 to 100 g L(-1) suspensions. Added phytase, in effect, increases water-extractable P content of manure and the risk of environmental P dispersion. As the bioavailability and ecological effect of IP6-P appear to be regulated not only by pH-controlled enzyme activity but also by the associated counterions, the differential protective effects of cations influence the accuracy of manure PHP fraction estimates and increase phytate resistance to enzymatic dephosphorylation that may lead to its persistence in manure.

Phosphorus-31 Nuclear Magnetic Resonance Spectral Assignments of Phosphorus Compounds in Soil NaOH–EDTA Extracts

Soil P composition can be conveniently determined in alkaline extracts using solution 31P nuclear magnetic resonance (NMR) spectroscopy, but spectral assignments are based on fragmentary literature reports of model compounds in various extraction matrices. We report solution 31P NMR chemical shifts of model P compounds, including inorganic phosphates, orthophosphate monoesters and diesters, phosphonates, and organic polyphosphates, determined in a standardized soil P extractant (0.25 M NaOH and 0.05 M EDTA). Signals from nucleic acids (DNA −0.37 ppm, RNA 0.54 ppm) and phospholipids (phosphatidyl choline 0.78 ppm, phosphatidyl serine 1.57 ppm, phosphatidyl ethanolamine 1.75 ppm) could be differentiated in the orthophosphate diester region, and were identified in a sample of cultured soil bacteria. Inorganic and organic polyphosphates could be differentiated by the presence of a signal at −9 ppm from the α phosphate of organic polyphosphates. Some orthophosphate diesters, notably RNA and phosphatidyl choline, degraded rapidly to orthophosphate monoesters in NaOH–EDTA although DNA, other phospholipids, and orthophosphate monoesters were more stable. Changes in probe temperature had a marked influence on signal intensities and the relative magnitude of signals from orthophosphate monoesters and inorganic orthophosphate, and we suggest that solution 31P NMR spectroscopy of soil extracts be performed at 20°C.

Polyvalent Cation Effects on -Inositol Hexa Dihydrogenphosphate Enzymatic Dephosphorylation in Dairy Wastewater

Information is needed on organic polyphosphates such as myo-inositol 1,2,3,5/4,6-hexakis dihydrogenphosphate or phytate (IP6) contribution to the sources and sinks of dissolved phosphorus (PO4–P) in the soil–manure–water system. Effects of Na+, Ca2+, Al3+, and Fe3+ and cation to IP6-P mole ratios on the enzymatic dephosphorylation of IP6 were studied to determine controlling mechanisms of dephosphorylation and persistence in manure. Phytate- and PO4–P were analyzed by high-performance liquid chromatography. Phytate dephosphorylation by Aspergillus ficuum (Reichardt) Henn. phytase EC 3.1.3.8 decreases by 50 ± 3.6 and 40 ± 4% at pH 4.5 and 6, respectively, as Ca2+ concentrations increase and cation to IP6-P mole ratios reach 6:6. Polyanionic IP6 has a high affinity for Al3+ and Fe3+ and reductions in dephosphorylation average 27 and 32% at a cation to IP6-P mole ratio of 1:6 for Al3+ and Fe3+, respectively, while reaching more than 99% at a mole ratio of 6:6. A phytase-hydrolyzable phosphorus (PHP) fraction is native to ruminant animal manure and is proportional to total solids (TS) concentration in 1 to 100 g L−1 suspensions. Added phytase, in effect, increases water-extractable P content of manure and the risk of environmental P dispersion. As the bioavailability and ecological effect of IP6-P appear to be regulated not only by pH-controlled enzyme activity but also by the associated counterions, the differential protective effects of cations influence the accuracy of manure PHP fraction estimates and increase phytate resistance to enzymatic dephosphorylation that may lead to its persistence in manure.

Oxygen-binding properties of total hemoglobin and isolated components of the terrestrial tortoise Geochelone carbonaria

The hemoglobin of the terrestrial tortoise Geochelone carbonaria was separated into two major components by ion exchange chromatography. The oxygen binding properties of both the unfractionated hemolysate and the separated components were investigated. The stripped hemoglobin of G. carbonaria displayed an oxygen affinity of approximately 6.3 mm Hg at pH 7.4 with a Bohr effect of about −0.55. The Hb-O 2 binding properties were also studied in the presence of the organic polyphosphates ATP and IHP. Inositol hexaphosphate (IHP) was more effective in lowering oxygen affinity, yielding a P 50 of 20.1 mm Hg at pH 7.4. The n 50 value were quite low in the stripped form of the protein (1.0–1.9) but increased in the presence of both organic phosphates, attaining a value of approximately 2.9 at pH 7.9. The oxygen binding properties of the major hemoglobin components were also investigated in the presence and absence of organic phosphates, and the values were compared with those obtained for the total hemolysate.

Mobile macromolecular carriers of ionic substances, 1. Transport of heavy metal ions by polymer analogues of teichoic acids

AbstractBioanalogous organic polyphosphates (PP) and polymer ion‐exchange membranes (IEM) were used to arrange a membrane system for the transportation of Zn(II), Mn(II), Cu(II), Co(II) and Ni(II) ions. The system consists of three membranes separating a feed and a strip solution with metal sulfates and sulfuric acid, respectively: Poly(trimethylene phosphate), α‐hydroxy‐ω‐(2‐ethylhexyl)poly(trimethylene phosphate), poly(1,2‐glycerol phosphate) and poly(2,3‐acetoxyglycerol phosphate) were used as ionic carriers for mimicking some transport functions of natural teichoic acids. These polyelectrolytes were found to facilitate the exchange‐diffusion process of heavy metal ions and to exhibit fluxes in the range 3,5 · 10−9 – 6 · 10−9 mol M(II)/(cm2 · s). The specific stationary fluxes typically change in the order JCu > JCo > JNi > JMn > JZn. The observed transport preference is consistent with the affinity order of teichoic acids or the bacteria cell wall toward Cu(II), Mn(II) and Zn(II).

Reexamination of the fractionation of total dissolved phosphorus in seawater using a modified UV-irradiation procedure, and its application to samples from Suruga Bay and Antarctic Ocean

Application of a modified UV-irradiation procedure to the fractionation of total dissolved phosphorus (TDP) in seawater was examined. By using a peristaltic pump, a sample solution was passed through a quartz column which was coiled around the three Hg-lamps and thus consistently exposed to a controlled UV-irradiation flux. The UV-flux was much lower than that historically employed to hydrolyze total dissolved organic phosphorus (DOP). Before and after the UV-irradiation, colorimetric measurements of soluble reactive phosphorus (SRP) were made by the method of Murphy and Riley (1962) to determine the liberation rate of orthophosphate from P-compounds in the sample. Experiments using 19 different P-compounds indicated that organic phosphate-esters containing only monomers of phosphate were readily decomposed by a 20-minute UV-irradiation period. Release of SRP from organic polyphosphates was minimal. The P released by this procedure is operationally referred to as “UV-P”. The procedure described herein was employed for fractionation of TDP in seawater. These studies defined several classes of dissolved P: (1) PO4−P(inorganic phosphate-P), (2) UV-P (Photodecomposable organic-P, most of which consists of organic monophosphate-P), and (3) Org-Poly-P (organic polyphosphate-P). Vertical profiles of DOP and UV-P observed in the oligotrophic regions of Suruga Bay and Antarctic Ocean indicated that UV-P was a major part of total dissolved phosphorus (TDP) in euphotic layers, where inorganic nutrients were probably limiting the active growth of phytoplankton. It is probable that UV-P can be utilized as a source of nutrient-P, in place of PO4−P.

Liquid membranes with organic polyphosphates as ionic carriers

AbstractPoly(hexylene phosphate) (1) and poly[poly(ethylene glycol)phosphate] (2) were examined as ionic carriers for concentration/separation processes, applying the liquid membrane technique. Experiments were performed with nickel and cobalt ions pertracted both from a single solution and bicomponent mixtures. 2, dissolved in 1,2‐dichloroethane, was found to be an effective carrier of nickel ions. The system seems to offer efficient separation of Ni(II)/Co(II) ions.

Dimer-tetramer transition in hemoglobins from Liophis miliaris—I. Effect of organic polyphosphates

1. 1. Hemoglobin from the water-snake Liophis miliaris in the stripped form presents high oxygen affinity of about P 50 = 1 mmHg and Hill coefficient of about 1.0 at pH from 6.8 to 8.5. 2. 2. In the presence of ATP such values become P 50 = 20 mmHg and n H about 2.0, respectively, at low pH from 6.5 to 7.5. 3. 3. When the pH increases an abrupt decrease of both P 50 and n H values occurs falling close to those found for the stripped hemoglobin. 4. 4. Gel-filtration in Sephadex G-100 equilibrated with 0.05 M Tris-HCl buffer containing 1 mM EDTA of the stripped hemoglobin show the presence of only one component of mol. wt of about 32,500 dt similar to the dimer of human hemoglobin A. 5. 5. The deoxy form of the dimer previously treated with ATP and placed on Sephadex column in the same condition but containing 1 mM IHP emerges as tetramer with mol. wt similar to that found for human hemoglobin, i.e. of about 65,000 dt. 6. 6. Results of the multiplicity of the snake hemoglobin, as well as the large alkaline Bohr effect in the presence of ATP previously reported, seems to be inconsistent due to the dimer-tetramer transition that occurs when ATP is bound to the stripped hemoglobin. 7. 7. A molecular mechanism involving the dimer-tetramer transition is proposed to described the oxygen transport in these animals.

Content of organic polyphosphates and their allosteric effects on haemoglobins from the water-snakes Helicops modestus and Liophis miliaris

1. 1. The ion-exchange Chromatographie analysis of the red cell nucleotides of two water-snakes with different degrees of adaptation to water showed a ratio ATP/GTP of about 2.3 in the more aquatic species Helicops modestus and of about 0.7 in Liophis miliaris. 2. 2. The haemoglobin oxygen affinity assayed in the presence of ATP, GTP, DPG and IHP in haemolysate from both snakes were determined as a function of pH. The Bohr effect values were always found to be increased in comparison with the stripped haemolysate, becoming about 1.5 times higher in L. miliaris compared with H. modestus by the nucleotide treatment. 3. 3. The Hill coefficient calculated from the slope of the haemoglobin binding curves was consistent with an increase in the protein cooperativity in the presence of the nucleotides assayed. The very low n H values of about 1.2 in absence of the effectors became close to 2.0 in their presence. 4. 4. The data of nucleotide Chromatographie analysis were compared to those found in fishes with bimodal respiration.

Characteristic features of the electronic and three-dimensional structure of organic phosphates and polyphosphates

Characteristic features of the electronic and three-dimensional structure of organic phosphates and polyphosphates

Competition in oxygen-linked anion binding to normal and variant human hemoglobins.

The two-state model of Monod, Wyman and Changeux has been used extensively in analysis of the functional behavior of human hemoglobin. Within the context of this model, the cooperativity, pH dependence of oxygen binding, and anionic regulation of oxygen affinity are all considered to be due to shifts in the equilibrium between low affinity (T) conformational states and high affinity (R) conformational states. The heterotropic effectors, such as protons, carbon dioxide, inorganic anions, and organic polyphosphates, are considered to be principally active in causing shifts in the allosteric equilibrium constant L. For both simple anions, such as chloride, and strong effectors, such as inositol hexaphosphate, the allosteric equilibrium constant L increases with increasing concentrations of the effector. Detailed examinations of the functional properties of human hemoglobin have revealed in recent years that the two-state model is a beautiful and elegant format for discussing conformational changes within a given condition, but that an infinite number of "T" and "R" states are, in fact, possible. In this paper, the consequences of the occurrence of specific oxygen-linked anion binding sites on the alpha and beta chains of the human hemoglobin tetramer are discussed in terms of the generalized two-state model. In particular, attention is directed to the interesting fact that the allosteric equilibrium constant can be decreased by addition of chloride instead of increased if the ligand binding behavior is analyzed in the presence of inositol hexaphosphate. The structural basis for competitive anionic effects involves the binding sites for anions on both alpha and beta chains. Analysis of the distinct effects of small and large anionic effectors is facilitated through studies of specific human hemoglobin variants and hemoglobins where the C terminal arginines of the alpha chains have been removed by ezymatic digestion.

Structure of the molecular orbitals in organic phosphates and polyphosphates

Structure of the molecular orbitals in organic phosphates and polyphosphates

The Reaction of Cyanate with the a and b Subunits in Hemoglobin: Effects of Oxygenation, Phosphates, and Carbon Dioxide

To evaluate the influence of oxygenation on the reactivity of hemoglobin to cyanate, oxy- and deoxyhemoglobin were carbamylatedwith a low (2.5 mM) concentration of [14C]-cyanate for up to 100 min. The radioactivity of the isolated tryptic peptides of purified carbamylated globin chains (αc and βc) was determined. Under these reaction conditions, 97% or more of the cyanate was bound to the NH2-terminal peptides of the α and β chains in oxy- as well as in deoxyhemoglobin. A modification of the ρ-chloromercuribenzoate method for chain separation was used to measure the rate of the reaction with cyanate of both types of chians in tetrameric hemoglobin. The reaction of hemoglobin with cyanate was strongly oxygen linked. the α chains were carbamylated twice as rapidly in deoxy- as compared to oxyhemoglobin in both phosphate and bis-tris buffers. The β chain carbamylation was less dependent upon oxygenation of hemoglobin in bis-tris buffer, and did not vary at all with oxygenation in phosphate buffer. Organic Polyphosphates (2,3-diphosphoglycerate and inositol hexaphosphate) inhibited β chain carbamylation selectively. The effect of pH on the carbamylation of α and β chains in deoxyhemoglobin was studied. At a fixed pH, CO2 decreased the reaction rate of α and β chain NH2 termini to the same extent.