Large Amounts Of Phosphate Research Articles

The identification and classification ofAcinetobacter strains exhibiting variations in phosphate accumulation by SDS-PAGE and numerical analysis

The limitation of phosphate concentrations in effluents is of international concern because of the risk of eutrophication.Acinetobacter spp. have often been isolated from activated sludge plants exhibiting enhanced phosphate removal. The ability to remove phosphate from mixed liquor byAcinetobacter isolates and reference strains was, therefore, determined. The ability to remove phosphate was found to vary among theAcinetobacter strains. The taxonomic relationships among these various strains were elucidated by SDS-PAGE and numerical analysis, and the correlation between their taxonomy and phosphate uptake ability was determined. The ability to remove large amounts of phosphate was found to be strain specific rather than species specific. The classification ofA. haemolyticus andA. baumannii as separate species is questioned.

Variation of Chemical Properties of Ground Water in Farm Land

Effect of the fertilizer on chemical properties of ground water was investigated from March to November in a farm land of West of Fukuoka City. The variation of dissolved oxygen (DO) and nitrate (NO3-N) concentrations in ground waters with time was greatly different by the surface conditions of the land: paddy field or vegetable field. In the case of the former, the DO and NO3-N concentrations decreased gradually from May and showed a minimum at the end of August. While, in the case of the latter, the concentrations were highly constant. Despite that the large amount of phosphate was manured, its concentration in ground waters was low.

Iron environment in ferritin with large amounts of phosphate, from Azotobacter vinelandii and horse spleen, analyzed using extended x-ray absorption fine structure (EXAFS)

The iron core of proteins in the ferritin family displays structural variations that include phosphate content as well as the number and the degree of ordering of the iron atoms. Earlier studies had shown that ferritin iron cores naturally high in phosphate, e.g., Azotobacter vinelandii (AV) ferritin (Fe:P ratio = 1:1.7), had decreased long-range order. Here, the influence of phosphate on the local structure around iron in ferritin cores is reported, comparing the EXAFS of AV ferritin, reconstituted ferritin [the protein coats of horse spleen ferritin mixed with Fe(II) with and without phosphate at pH 7] (Fe:P ratio = 1:0.25), and native horse spleen ferritin (Fe:P ratio = 1:0.125); reconstituted horse spleen ferritin without phosphate was indistinguishable from native horse spleen ferritin (HSF) in the analysis. In contrast, when the phosphate content was high in AV ferritin and horse spleen ferritin reconstituted with phosphate, the average iron atom had five to six phosphorus neighbors at 3.17 A. Moreover, the number of detectable iron neighbors was lower when phosphate was high or present during reconstitution (2-3 vs 5-6), and the interatomic distance was longer (3.50 vs 3.03 A), indicating that some phosphate bridges neighboring iron atoms. However, the decrease in the number of detectable iron-iron neighbors compared to HSF and the higher number of Fe-P interactions relative to Fe-Fe interactions suggest that some phosphate ligands were chain termini, or blocked crystal growth, and/or introduced defects which contributed both to the long-range disorder and to altered redox properties previously observed in AV ferritin.

Isotopically exchangeable phosphate in freshwater sediments: Effects of u.v.-irradiation, formaldehyde, solid/solution ratio, and pH on its experimental determination

The influence of bioactivities, solid/solution ratio and the pH on the isotopic exchangeability of phosphate in a freshwater sediment was investigated. From the comparison of the results obtained for the same sample in the presence or absence of formaldehyde, it is concluded that microorganisms can effect the analysis for isotopically exchangeable phosphate. Irradiation with u.v.-light caused a sharp rise in isotopic exchangeability. In the pH-range 6.6–8.4 isotopic exchangeability of phosphate increases with decreasing pH-value which is attributed to an easier exchange of H 2PO 4 − than of HPO 4 2−. The influence of the solid/solution ratio on the isotopic exchangeability of phosphate in the solid phase is small or nil. However, because of the relatively large amount of phosphate that goes into solution, the total isotopic exchangeability E i of phosphate in the solid and liquid phase together is strongly increased at a low solid/solution ratio. From these results it is concluded that in order to make a meaningful comparison of isotopically exchangeable phosphate in different soils or sediments it is essential to work at a nearly constant pH and solid/solution ratio. Such a comparison was made for 26 freshwater sediments from the Rhine/Meuse delta, in the presence and absence of 0.17 mol l −1 formaldehyde as a biological inhibitor and it is concluded that addition of the latter is essential. The lowest total isotopic exchangeabilities of phosphate, (15–25) were measured in the sediments collected from the Haringvliet, whereas higher values (40–80) were found in the sediments from the Brielse Meer and the Grote Rug. This could well be indicative of a similar variance in the biological availability of the phosphate in the investigated sediments.

The effect of phosphate detergent bans on consumer clothing expenditures

Metropolitan sales data from the 1977 Census of Retail Trade were analysed to test whether phosphate detergent bans increased consumer expenditure on clothing: positive effects were found for men's clothing and domestic fabrics (sheets and towels). In areas with medium water hardness the average annual cost in 1977 equalled $11·08 per household. For 1984, the equivalent cost was $14·17 per household. These results are consistent with laboratory studies that show decreased detergency associated with non‐phosphate detergents and increased fabric wear associated with carbonate‐built detergents, the principle substitute for phosphate detergents. The bans impose the largest cost on hard water areas. In both ban and non‐ban areas in 1977 higher laundering costs were associated with higher water hardness. Analysis of 1972 data indicated that water hardness did not affect clothing expenditures significantly when detergents contained large amounts of phosphates.

Surface properties and equilibrium kinetics of hydroxyapatite powder near the solubility equilibrium

Abstract The behavior of high specific surface area synthetic hydroxyapatite (HA) powder has been studied for solutions of different compositions near the solubility equilibrium in order to avoid pure dissolution or remineralization processes. By following the evolution with time of pH and the concentrations of calcium and phosphate in solution as a function of time, ion exchanges between the surface and the solution were studied up to the attainment of the solubility equilibrium. The solubility product (Ca 2+ ) 5 (PO 4 3− ) 3 (OH − ) in the presence of 8 × 10 −2 M KCl was found to be 1.189 × 10 −60 at 37°C. The kinetics observed suggest two mechanisms. During a very short time H + and Ca 2+ ions are rapidly desorbed from the surface, then a slower desorption or readsorption process occurs leading to equilibrium. Large amounts of phosphate and calcium ions are able to be adsorbed onto the surface and regulate the attainment of this equilibrium. It is also shown that the HA surface is amphoteric and acts as a buffer in a rather large pH range. The presence of 0.23% carbonate in the samples does not seem to play any role since the kinetics of carbonate release indicate that this trace element is incorporated into the powder rather than adsorbed on the surface.

Ionic adsorption properties and equilibration kinetics of biological enamel powder near thermodynamic equilibrium

The behaviour of biological enamel powder (EP) has been studied for solutions of different compositions close to solubility equilibrium in order to avoid pure dissolution or remineralization reactions. Surface ion exchange kinetics have been determined up to solubility equilibrium by continuous monitoring of the solution H +, Ca 2+ and PO 3- 4 ions versus time. The solubility product K s, (Ca 2+) 5(PO 3- 4) 3(OH -), was found to be 2.51x10 -61 at 37° C in the presence of 8x10 -2 M KCI, whereas K s decreased when the KCl concentration in the solution decreased. Often kinetics suggested the combination of two mechanisms. A rapid proton release and/or calcium ion release or uptake was followed by slow readsorption or desorption processes leading to the establishment of equilibrium. Large amounts of phosphate and calcium ions are adsorbed onto the surface and regulate the phenomenon. No influence of the 4.42% carbonate ions incorporated in the samples was detected in the equilibrium kinetics close to the solubility limits.

Phosphate Content of Human Amniotic Fluid and Its Relationship to Bacterial Growth Inhibition

A sensitive method for assay of phosphate has been adapted for use with human amniotic fluid. The presence of an antibacterial factor in amniotic fluid inactivated by phosphate suggested that measurement of free phosphate may predict antibacterial activity and hence susceptibility to intrapartum infection. The intrinsic antibacterial activity of amniotic fluid was altered by addition of large amounts of phosphate, but the intrinsic concentration of free phosphate correlated poorly with antibacterial activity and organically combined phosphates did not appear to be related to the antibacterial property.

Ordered phosphorylation of 40S ribosomal protein S6 after serum stimulation of quiescent 3T3 cells.

The amino acids and tryptic peptides that become phosphorylated in 40S ribosomal protein S6 after serum stimulation of quiescent 3T3 cells were examined by two-dimensional thin-layer electrophoresis. In the maximally phosphorylated form of the protein, most of the phosphate was incorporated into serine and a small amount, into threonine. Digestion of this form of the protein with trypsin revealed 10 major phosphopeptides. All 10 contained phosphoserine and 2 of the 10 also contained phosphothreonine. Next, the five forms of increasingly phosphorylated S6 were individually separated on two-dimensional polyacrylamide gels or total S6 was isolated from cells that were stimulated for only a short time and their phosphotryptic maps were analyzed. The results showed that, as larger amounts of phosphate were added to S6, the phosphopeptides appeared in a specific order.

PHOSPHATE ADSORPTION IN ALLOPHANIC SOILS

SummaryThe kinetics and heats of phosphate adsorption were measured on the <2 μm Na‐saturated fractions of three allophanc‐rich soils from Japan. Between 50 and 2250 μmol P g−1 as sodium phosphate were added to the soil fractions at pH 5 and pH 7, and at initial concentrations of 5 and 25 × 10−4m to avoid aluminium phosphate precipitation.An initial ‘instantaneous’ adsorption associated with exposed sites and, simultaneously, two inverse exponential rates of adsorption on internal and freshly forming external sites were observed. These rates are attributed to changes in the microstructure of allophane and to the desorption of organic matter held on allophanic surfaces. This interpretation is strongly supported by corresponding changes in the heats of adsorption with time. Calorimetry clearly indicates that when very large amounts of phosphate are added, new and very reactive surfaces are progressively exposed.More phosphate was adsorbed when the soil was acid and when the soil contained less organic matter.

Reevaluation of properties of acetyl-CoA carboxylase from rat liver.

Rat liver acetyl-CoA carboxylase can be rapidly isolated by a new procedure which uses avidin-Sepharose affinity chromatography. The isolated enzyme has Mr = 260,000; none or very little of the proteolytic products of the carboxylase which are formed in conventional purification procedures are found in our preparations. It is apparent that the previously reported subunit of the carboxylase, with Mr = 230,000, is itself the product of proteolysis. The properties of the enzyme produced by our new method are quite different from those of the conventionally prepared enzyme. Our enzyme contains 6 mol of alkali-labile phosphate/mol of subunit, rather than 2 mol; the Km for acetyl-CoA is about 8-fold higher and the specific activity is only about one-fifth of that previously reported. The large amount of phosphate does not appear to cause the low specific activity of the new enzyme preparation, because alkaline phosphatase treatment reduces the number of phosphates/subunit from 6 to 3 mol but does not change the specific activity.

Relationship between phosphate content and serological activities of the mannans of Candida albicans strains NIH A-207, NIH B-792, and J-1012

The mannans from Candida albicans strains NIH A-207 (serotype A), NIH B-792 (serotype B), and J-1012 (serotype C) were fractionated on a column of diethylaminoethyl-Sephadex into five subfractions containing different amounts of phosphate. Antibody-precipitating activities of the mannan subfractions of strains NIH A-207 and NIH B-792 were proportional to their phosphate content, while those of strain J-1012 did not show regularly proportional precipitin activity. A similar tendency was also observed in the cross-reaction between the mannan su,fractions of strains NIH A-207 and J-1012 and their heterologous antisera. The mannans of strain NIH B-792 showed lower cross-reactivities against antisera of strains NIH A-207 and NIH B-792, i.e., only two subfractions containing larger amounts of phosphate were able to react with these antisera.

Purification and physicochemical properties of fatty acid synthetase and acetyl-CoA carboxylase from lactating rabbit mammary gland.

Fatty acid synthetase was purified 13-fold from lactating rabbit mammary glands by a procedure which involved chromatography on DEAE-cellulose, ammonium sulphate precipitation and gel filtration on Sepharose 4B. The preparation was completed within two days and over 100 mg of enzyme was isolated from 100--150 g of mammary tissue, which represented a yield of over 40%. The preparation was homogeneous by the criteria of polyacrylamide gel electrophoresis and ultracentrifugal analysis. The sedimentation constant, S20,w was 13.3 S, the absorption coefficient, A280nm1%, measured refractometrically was 10.0 +/- 0.1, and the amino acid composition was determined. The subunit molecular weight determined by gel electrophoresis in the presence of sodium dodecyl sulphate was 252,000 +/- 6,000, and the molecular weight of the native enzyme measured by sedimentation equilibrium was 515,000. These experiments indicate that at the concentrations which exist in mammary tissue (2--4 mg/ml) fatty acid synthetase is a dimer. The purified enzyme did however show a tendency to dissociate to a monomeric 9-9S species on storage for several days or following exposure to a low ionic strength buffer at pH 8.3. There was only a small quantity of alkali labile phosphate (0.2 molecules per subunit) bound covalently to the purified enzyme. Acetyl-CoA carboxylase was purified 300-fold in a 50% yield within 24 h by ammonium sulphate and polyethylene glycol precipitations [Hardie, D.G. and Cohen, P. (1978) FEBS Lett. 91, 1--7]. The preparation was in a state approaching homogeneity as judged by polyacrylamide gel electrophoresis, gel filtration on Sepharose 4B and ultracentrifugal analysis. The sedimentation constant, S20,w, was 50.5 S, the absorption index, A280nm1%, was 14.5 +/- 0.7, and the amino acid composition was determined. The subunit molecular weight of acetyl-CoA carboxylase determined by gel electrophoresis in the presence of sodium dodecyl sulphate was identical to that of fatty acid synthetase (252,000) as shown by electrophoresis of a mixture of the two proteins. The preparations also contained two minor components of molecular weight 235,000 and 225,000, which appear to be derived from the major species of mol. wt 252,000. A large emount of phosphate (3.2 molecules per subunit) was found to be bound covalently to the purified enzyme. The properties of fatty acid synthetase and acetyl-CoA carboxylase are compared to those obtained by other workers.

The stoicheiometry of the absorption of protons with phosphate and L-glutamate by yeasts of the genus Saccharomyces

1. A study was made of the pH changes occurring when 0.1-4 mumol of glutamate, phosphate and certain phosphate esters was added at about pH 4.8 to washed cell preparations (50 mg dry wt.) of strains of Saccharomyces. The system also contained deoxyglucose and antimycin to inhibit energy metabolism and so prevent proton ejection from the yeast. 2. A strain of Sacc. carlsbergensis was grown in a chemostat with a limiting supply of phosphate in order to enhance the subsequent rate of phosphate transfer into the yeast. These preparations absorbed 0.2 mumol of phosphate with about 3 equiv. of protons/mol of phosphate. The charge balance was maintained by the efflux of 2 equiv. of K-+ from the yeast. 3. Larger amounts of phosphate were absorbed with fewer proton equivalents. 4. Arsenate and phosphate caused similar pH changes. 5. Glucose 6-phosphate, ATP and certain order phosphate esters each initiated a rise in pH, possibly because hydrolytic extracellular enzymes released phosphate that was subsequently absorbed. 6. Four strains of yeast were grown with glutamate as principal source of nitrogen. Each absorbed extra protons in the presence of L-glutamate. 7. One of them, a strain of Sacc. cerevisiae, absorbed 0.2 mumol of glutamate with 3equiv. of protons/mol of glutamate, and in these circumstances 1-2 equiv. of K-+ left the yeast cells. 8. The role of ionic gradients in the transport of these anions is discussed.

The Influence of Phosphate Uptake on Cation Uptake in Fusarium oxysporum f. sp. vasinfectum

Abstract Fusarium oxysporum grown in a low phosphate medium was found to take up several times as much K from KH2PO4 as from KCI solutions. Large amounts of phosphate also were taken up from KH2PO4. Similar large uptakes of Na and phosphate took place from solutions of NaH2PO4. Substantial quanties of phosphate were taken up from solutions of Ca(H2PO4)2 in the absence of any appreciable Ca uptake. When the fungus was grown in a medium containing high phosphate, little or no uptake of phosphate from KH2PO4 solutions occured and the K Uptake was at the same level as from KCI solutions. During large phosphate uptake sizable reductions in the organic acid content of the fungal cells were observed. Much, but not all, of the data could be explained on the basis of maintenance of charge balance within the cells. – The respiratory rate of fungus, grown in a low P medium, was markedly increased in KH2PO4 solution. Fungus, grown in a medium with high phosphate, had a higher respiratory rate which showed only a slight response to KH2PO4 solution. Fungus, grown in a medium with high phosphate, had a higher respiratory rate which showed only a slight response to KH2PO4.

The spectrophotometric determination of anions by solvent extraction with metal chelate cations. part XX : Trichloroacetic acid with tris(1,10-phenanthroline)iron(ii) chelate

Trichloroacetic acid can be extracted from an aqueous solution by nitrobenzene with tris(1,10-phenanthroline)iron(II) chelate, and can be determined spectrophotometrically by measuring the extract at 516 nm. The extracted species is probably [Fe(phen) 3].(CCl 3COO) 2. Beer's law is obeyed over the concentration range 1.0·10 -5–1.0·10 -4 M trichloroacetic acid in aqueous solution. Large amounts of phosphate and sulfate and moderate amounts of chloride, acetic acid, and monochloroacetic acid do not interfere, equal amounts of dichloroacetic acid give a slight positive error

Effective separation of condensed phosphate by ion-exchange chromatography

A conventional method of ion-exchange chromatography has been modified effectively for separating ortho- and pyro-phosphates, which were contained as impurities in recrystallized tripoly-phosphate, by gradient elution with 500 ml of 2×10-3 N HCl, to which 0.6 M KCl is added continuously and mixed. The separation can be brought about at an elution volume of 100ml for orthophosphate, of 200 ml for pyrophosphate and of 300 to 350 ml for tripolyphosphate. In the course of pretreatment with molybdic acid for colorimetry, a faint yellow coloration is developed due to phosphomolybdate, if 10 ml of the fractionated aliquot contains a large amount of phosphate. One tenth portion of the yellow solution is then diluted to 10 ml for the subsequent colorimetry by the molybdenum blue method, in order to obtain an appropriate range of optical density. The use of 25 cm length of resin column allows to take some hundred milligrams of sample tripolyphosphate for the determination of the phosphates as impurities as low as 0.01%. For the analysis of higher condensed polyphosphate, an eluent of more concentrated KCl solution is employed, for instance 1 M or 2 M KCl gives better separation similarly to that observed with diluted eluent. A recovery of total phosphorous is more than 95% by colorimetry.

Separation and determination of sulphate in large amount of phosphate by ion-exchange chromatography

多量のリン酸塩中に含まれる硫酸イオンの分離に,塩素形強塩基性陰イオン交換樹脂を利用した.0.1N塩酸-0.19N塩化カリウム溶離液でリン酸のみを先に溶離し,ついで2N塩化カリウム溶離液で硫酸イオンを溶離し,これをキレート滴定法で定量した.

Studies of the relative importance of iron and aluminium in the sorption of phosphate by some Australian soils

Phosphate sorption by 47 acid surface soils was correlated with the amounts of iron and aluminium extracted from them by ammonium oxalate at pH 3.2. The correlation coefficients, when all soils were considered, were highest for sorption and aluminium. For certain groups of soils, iron or iron plus aluminium gave higher coefficients than did aluminium. The relative contributions of iron and aluminium to phosphate sorption could not be accurately assessed from these correlation coefficients because iron and aluminium themselves were positively correlated. Eleven of the forty-seven soils were chosen for determinations of the effect of selective removal of iron and aluminium on their ability to sorb phosphate. Soils usually sorbed less phosphate after various amounts of their iron and aluminium had been extracted chemically. Many of them, however, still sorbed large amounts of phosphate in spite of the removal of reducible iron and acid-soluble aluminium. Soils and haematite extracted with sodium dithionite in acetate at pH 5.0 usually sorbed more phosphate than untreated samples. This was due largely to the incomplete removal of reduced iron and could be prevented by washing with dilute acid. Acid washing after dithionite treatment caused sorption by the soils to fall below the values for the untreated samples and virtually destroyed that of the oxide. It also removed much aluminium from the soils. The amount by which sorption fell below that of the untreated samples could be accounted for largely by this aluminium. It was concluded tentatively that, for most of the soils, phosphate sorption was due to acid-soluble aluminium and to sorption sites which were non-reducible and insoluble in dilute acid. The contribution of reducible iron in the original soils remains in doubt, but it could be a minor one.

Die Bestimmung von Fluor in Mineralwässern mittels Zr-Xylenolorange

The method proposed, based on the decolorisation of the zirconium-Xylenol Orange chelate in a 1.2 N hydrochloric acid medium, permits rapid determination of fluorine in mineral waters and other materials containing relatively large amounts of phosphate, sulphate, aluminium, iron III and other ions.