Phosphate Released Research Articles

Phospate Release Study on Silica Gel and Amino Silica Hybrid Sorbent from Lapindo Mud

This study discusses the rate of phosphate release in Silica Gel (SG) and Amino Silica Hybrid (ASH) sorbents using Lagergren pseudo-first Order Kinetic and Lagergren Pseudo-second Order Kinetic. ASH synthesis was done by adding a 3-aminopropyltriethoxysilane (APTES) modifier to sodium silicate from Lapindo Mud. Phosphate binding was carried out at 30 minutes with a stirring speed of 250 rpm. Analysis using IR spectrophotometer showed the success of ASH synthesis as indicated by the appearance of functional groups -NH2 and -CH2 at wavenumbers 1635 cm-1 and 1473 cm-1. With the continuous method, SG and ASH sorbents that have bound phosphate were then desorbed at various times of 2, 3, 4, 7, 8, and 9 days. The amount of phosphate anion released from SG and ASH was tested using a UV-Vis instrument then modeling was carried out to determine the rate of phosphate release. The desorption results showed the amount of phosphate released from the ASH sorbent was less than that of the SG sorbent. Through modeling, the proceeds of the phosphate release rate follow the Lagergren Pseudo-second Order Kinetic with an experimental QE of 0.22089 M/g for SG sorbent and 0.33333 M/g for ASH sorbent.

Assaying Paenibacillus alvei CsaB-Catalysed Ketalpyruvyltransfer to Saccharides by Measurement of Phosphate Release.

Ketalpyruvyltransferases belong to a widespread but little investigated class of enzymes, which utilise phosphoenolpyruvate (PEP) for the pyruvylation of saccharides. Pyruvylated saccharides play pivotal biological roles, ranging from protein binding to virulence. Limiting factors for the characterisation of ketalpyruvyltransferases are the availability of cognate acceptor substrates and a straightforward enzyme assay. We report on a fast ketalpyruvyltransferase assay based on the colorimetric detection of phosphate released during pyruvyltransfer from PEP onto the acceptor via complexation with Malachite Green and molybdate. To optimise the assay for the model 4,6-ketalpyruvyl::ManNAc-transferase CsaB from Paenibacillus alvei, a β-d-ManNAc-α-d-GlcNAc-diphosphoryl-11-phenoxyundecyl acceptor mimicking an intermediate of the bacterium’s cell wall glycopolymer biosynthesis pathway, upon which CsaB is naturally active, was produced chemo-enzymatically and used together with recombinant CsaB. Optimal assay conditions were 5 min reaction time at 37 °C and pH 7.5, followed by colour development for 1 h at 37 °C and measurement of absorbance at 620 nm. The structure of the generated pyruvylated product was confirmed by NMR spectroscopy. Using the established assay, the first kinetic constants of a 4,6-ketalpyuvyl::ManNAc-transferase could be determined; upon variation of the acceptor and PEP concentrations, a KM, PEP of 19.50 ± 3.50 µM and kcat, PEP of 0.21 ± 0.01 s−1 as well as a KM, Acceptor of 258 ± 38 µM and a kcat, Acceptor of 0.15 ± 0.01 s−1 were revealed. P. alvei CsaB was inactive on synthetic pNP-β-d-ManNAc and β-d-ManNAc-β-d-GlcNAc-1-OMe, supporting the necessity of a complex acceptor substrate.

Stannous Fluoride Preventive Effect on Enamel Erosion: An In Vitro Study.

The aim of this in vitro study was to evaluate the effects of a single dose application of two daily toothpastes on enamel exposed to acid attack. The research was conducted on human molars enamel fragments (n = 72). The two different toothpastes active ingredients were sodium fluoride (NaF) and stannous fluoride (SnF2). They were compared in protecting the surface of the enamel exposed to three acids: citric acid, lactic acid and hydrochloric acid. A spectrophotometer was used to measure the calcium ions and phosphate released in the solutions by the enamel specimens. Afterward, ionic concentrations were analyzed through the t-Student test, in order to estimate the significance level (p < 0.05) of the solubility differences obtained between the treatment and control groups. Finally, sample surfaces were analyzed with scanning electron microscopy and X-ray energy dispersive spectroscopy (SEM/EDX). The two analyzed toothpastes did not reveal any statistically significant variation in the release of calcium and phosphate (p > 0.05). Nevertheless, acid-resistant deposits were detected in samples treated with stannous fluoride and exposed to lactic acid, though the presence of tin ion deposits on samples treated with stannous fluoride was not shown. A single dose of a fluoride-based toothpaste before different acids attack, in simulated oral cavity conditions, did not show a significant preventive effect.

The Study of Phosphate Release from Artificial Sediment into Water Body Using Diffusive Gradient in Thin Film (DGT) Device in Oxic Condition

The phenomenon of phosphate release in sediments into water bodies under oxic environment has been investigated using the Diffusive Gradient in Thin Film (DGT) technique. This research consists of several stages: polymer synthesis and DGT probe assembly, sediment sampling, DGT deployment in oxic conditions, and phosphate analysis from DGT adsorption results. Acrylamide polymer was successfully synthesized with a composition 15% acrylamide; N-N'-methylenebisacrylamide 0.1% and ferrihydrite as binding gels. DGT probes were assembly by placing a 16 x 3.2 cm polyacrylamide gel, binding gels and filter membranes on the DGT probes. The sediment sample was taken from the Bogor Botanical Gardens at the coordinates 6°36’00.6” S; 106°47’51.0” E. The DGT probe was placed in sediment samples for 1, 3 and 7 days in oxic conditions. After the prescribed time, the binding gel was removed and cut every 1 cm depth, then eluted using 0.25 M H2SO4 and the phosphate concentration was measured using spectrophotometry method. The results showed that the phosphate concentration tends to be higher with the increasing incubation time and depth. Maximum CDGT phosphate released on day 1, day 3 and day 7 were 1.00 µg/L at a depth of 14 cm, 6.61 µg/L at a depth of 14 cm, and 20.92 µg/L at a depth of 11 cm respectively. This ensures that the phosphate in water bodies comes from biogeochemical processes that occur in sediments and is successfully measured through DGT techniques.

Model-Based Analysis of Reactive Transport Processes Governing Fluoride and Phosphate Release and Attenuation during Managed Aquifer Recharge.

In water-scarce areas, the reclamation of wastewater through advanced water treatment and subsequent reinjection into depleted aquifers is an increasingly attractive water management option. However, such injection can trigger a range of water-sediment interactions which need to be well understood and quantified to ensure sustainable operations. In this study, reactive transport modeling was used to analyze and quantify the interacting hydrogeochemical processes controlling the mobilization of fluoride and phosphate during injection of highly treated recycled water into a siliciclastic aquifer. The reactive transport model explained the field-observed fluoride and phosphate transport behavior as a result of the incongruent dissolution of carbonate-rich fluorapatite where (i) a rapid proton exchange reaction primarily released fluoride and calcium, and (ii) equilibrium with a mineral-water interface layer of hydrated dibasic calcium phosphate released phosphate. The modeling results illustrated that net exchange of calcium on cation exchange sites in the sediments postbreakthrough of the injectant was responsible for incongruent mineral dissolution and the associated fluoride and phosphate release. Accordingly, amending calcium chloride into the injectant could potentially reduce fluoride and phosphate mobilization at the study site. Insights from this study are broadly applicable to understanding and preventing geogenic fluoride mobilization from fluoride-bearing apatite minerals in many other aquifers worldwide.

Effect of Salinity and Oxygen Condition on Phosphate Release from Marine Sediment Measured Using Diffusive Gradient in Thin Film (DGT) Technique

The diffusive gradient in thin film (DGT) is an analytical technique to determine phosphate in the environment. This technique uses a thin film diffusive hydrogel in contact with a binding phase (ferrihydrite) to binding of phosphate. The released phosphate from marine sediment of Jakarta Bay was studied by DGT technique for effect of salinity and oxygen condition. Effect of salinity was observed by NaCl concentration from 0-35 g/L. The maximum phosphate release from sediment was occurred at the concentration of NaCl 30 g/L with incubation for 15 days with phosphate released at 113.99 μg/L, MDGT of 4.7723 μg and CDGT of 17.56 μg/L. The experiment showed the increase of MgCl2 and CaCl2 concentration reduced phosphate release from sediment. The condition of oxygen indicating the release of phosphate under anaerobic conditions is greater than the aerobic condition. The aerobic conditions with incubation for 21 days showed the release of phosphate from sediment to overlying water of 124.72 μg/L, MDGT of 2.4492 μg and CDGT of 6.4380 μg/L. Anaerobic conditions with incubation for 21 days showed phosphate release from sediment to overlying water of 208.62 μg/L, MDGT of 6.1081 μg and CDGT of 16.06 μg/L. The experiment shows that salinity and oxygen concentration influences phosphate release from marine sediment of Jakarta Bay.

Effects of floating Azolla on phosphorus fluxes and recovery from former agricultural lands in wetland microcosms

ABSTRACT The long-term fertilization results in accumulation of phosphorus especially in the top layer of the soils. Inundation of agricultural lands leads to a switch to anaerobic soil condition, causing reduction of iron and leaching of phosphate simultaneously. From the ecological and environmental perspective, high nutrients flux especially phosphorus will increase the possibility of eutrophication in aquatic system. The fern Azolla had a good potential to adsorb phosphorus, it also has distinctive nitrogen-fixing capacity. We conducted a 10-week aquarium experiment to investigate the phosphorus release capacity from two agricultural soils in the Netherlands with different Fe and P concentrations but comparable Fe/P ratios. Besides, the research questions rose to whether Azolla could use the mobilized phosphate released from the soils for growth. We also tried to find an effective indicator to estimate the actually phosphate mobilization from sediment to water layer. Results showed that the soils with high Fe and P concentrations had higher phosphate release rate compared with the soil with low Fe and P concentrations. Pore water Fe: PO4 3− ratios were valid to identify P release to surface water, when the Fe: PO4 3 − ratios less than 8 mol mol−1 substantial phosphorus mobilization occurred. The conclusions showed that the actual mobilization of phosphate is more important than the phosphorus retained in the sediments for the internal PO4 3 − fluxes. From 10-week experimental results, we found that Azolla can reuse the phosphate retained in soils thus removed the mobilized phosphate in a moderately low surface water nutrient loading.

Phosphate oxygen isotope evidence for methylphosphonate sources of methane and dissolved inorganic phosphate

The ocean is an important source of methane, however, the sources of oceanic methane and mechanisms of its release to the atmosphere have only recently begun to be understood. Recent studies have identified methylphosphonate (MPn) as a previously unknown and likely source of methane in the aerobic ocean (Karl et al., 2008), as well as shown the biosynthesis of methylphosphonic acid to be a widespread trait in marine microbes (Metcalf et al., 2012). The mechanisms and reaction pathways from MPn to free methane, however, have not been well studied. Here we present results of laboratory studies on the photo-degradation of MPn, a likely mechanism of methane release to the atmosphere and phosphate release to the surface oceans. Phosphonoacetic acid was also studied as an additional model compound for comparison. We used the multi-labeled water isotope probing (MLWIP) approach, involving 18O-labeled waters to probe the photolytic mechanism of CP bond cleavage in phosphates through analysis of P released from MPn as PO4. These studies identified distinct reaction pathways involving phosphates compared with other common organophosphorus compounds (e.g., phosphoesters), as well as suggest the involvement of both ambient water and atmospheric oxygen in CP bond cleavage. There is only a small amount of water oxygen incorporated into product PO4 after cleavage of the CP bond in MPn, suggesting atmospheric O2 or radicals formed from O2 under Ultra Violet Radiation (UVR), as the primary source of O that replaces C in the CP bond of MPn. Model calculations suggest that the δ18OP signature of phosphate released via UV-degradation of phosphates is largely (75%) inherited from the original phosphate substrate. This opens up the possibility of tracing and differentiating specific phosphate sources of dissolved phosphate from other organophosphorus (Porg) sources (e.g., phosphoesters) used in primary production, as well as for tracing specific MPn sources of atmospheric methane.

Adsorption of phosphate by the cellulose-based biomaterial and its sustained release of laden phosphate in aqueous solution and soil

A new quaternary ammonium anion exchanger derived from the cellulose-based biomaterial (sugarcane bagasse) was prepared for phosphate adsorption and then the phosphate saturated bio-sorbent was used as a slow-release phosphate fertilizer in water or soil. Results indicated that the adsorption of phosphate by the quaternary ammonium based sugarcane bagasse (QA-SB) was increased with the increase of temperature, and the adsorption was optimal at pH of 4.33–5.88. The capture of phosphate by the QA-SB in fixed-bed column was calculated to be 18.9–21.4 mg/g in the background of Cl− or NO3− (100 mg/L). In particular, the amount of laden phosphate was significantly decreased to 9.1 mg/g in the presence of coexisting SO42− (100 mg/L). As a result, a stronger competitve effect of SO42− on phosphate capture was observed as compared with those of Cl−, and NO3−. Results also indicated that the laden phosphate on the QA-SB could be desorbed efficiently and the recovery of phosphate onto the regenerated QA-SB still attained about 94.5% after 4 cycles. The slow-release of laden phosphate in aqueous solution could be continuously conducted for 12 d, with 43.6% of laden phosphate (11.6 mg/g) released. In contrast, phosphate (7.4 mg/g) released in soil reached equilibrium within 4 d, accounting for only 27.8% of laden phosphate released.

Study of phosphate release from Bogor botanical gardens’ sediment into pore water using diffusive gradient in thin film (DGT)

Eutrophication is one of the environmental problems caused by the excessive nutrients in aquatic ecosystems. In most lakes, phosphate is a limiting nutrient for algae photosynthesis. Even though the concentration of phosphate from external loading into the water body has been reduced, eutrophication could still be occured due to internal mobilization of phosphate from the sediment pore water into the overlying water. Therefore, the released phosphate from sediments and their interaction in the pore water must be included in the monitoring of phosphate concentration in aquatic system. The released phosphate from sediment into pore water has been studied by DGT device with ferrihydrite as binding gel and N-N′-methylenebisacrylamide as crosslinker. The results showed that DGT with 15% acrylamide; 0.1 % N-N′-methylenebisacrylamide and ferrihydrite as binding gel was suitable for the measurement of the released phosphate from sediment into pore water. The result of the deployed DGT in oxic and anoxic conditions in seven days incubation showed the released phosphate process from the sediment into pore water was affected by incubation time and the existence of oxygen in the environment. The released phosphate from the sediment into pore water in anoxic condition has a higher value than oxic condition. The experimental results of the deployed DGT in natural sediment core at a depth of 1 to 15 cm from the surface of the water for 7 days showed that the sediment has a different phosphate mass profile based on depth. The concentration of phosphate tends to be increased with depth. The maximum CDGT of phosphate released in oxic and anoxic conditions at 7th day period of incubation are 29.23 μg/L at 14 cm depth and 30.19 μg/L at 8 cm depth, respectively.

Photo-induced Phosphate Release from Organic Phosphorus Decomposition Driven by Fe(Ⅲ)-oxalate Complex in Lake Water

The phosphate released from organic phosphorus photo-decomposition has a significantly influence on the phosphorus levels in the water column in lakes. In order to reveal the effect of organic phosphorus photo-decomposition on phosphate level in lake water, the phosphate released from organic phosphorus photo-decomposition driven by Fe(Ⅲ)-oxalate complex under UV-Vis and sunlight irradiation was investigated in natural lake water using glyphosate as the model organic phosphorus. The effects of pH and initial concentration of Fe(Ⅲ), oxalate and glyphosate on the phosphate released from glyphosate photolysis were studied. The results showed that phosphate could be released from glyphosate degradation by Fe(Ⅲ)-oxalate complex under UV-Vis and sunlight irradiation. The concentration of phosphate reached 0.25 mg·L-1 and 0.18 mg·L-1 under UV-Vis and sunlight irradiation for 60 and 720 min, respectively. The amount of phosphate released increased with the increase of the initial concentration of Fe(Ⅲ), as well as the increasing oxalate and glyphosate concentration in lake water. However, the increase of pH could significantly inhibit this process in the reaction system. The concentration of phosphorus decreased with the addition of isopropanol, which indicated that the hydroxyl radical (·OH) was one of the main active oxygen species of Fe(Ⅲ)-oxalate complex. The rates of·OH production for Fe(Ⅲ)-oxalate/UV-Vis and Fe(Ⅲ)-oxalate/sunlight systems were 0.52×10-2 μmol·(L·min)-1 and 0.03×10-2 μmol·(L·min)-1, respectively. The steady-state concentrations of hydroxyl radical (·OH) for the Fe(Ⅲ)-oxalate/UV-Vis conditions were 4.74×10-16 mol·L-1 and 0.27×10-16 mol·L-1 for the Fe(Ⅲ)-oxalate/sunlight system.

The role of Fe(III) on phosphate released during the photo-decomposition of organic phosphorus in deionized and natural waters

The photo-decomposition of organic phosphorus is an important route for the phosphorus cycle by which phosphate is regenerated in the aquatic environment. In this study, the role of Fe3+ as a natural photosensitizer toward the decomposition of organic phosphorus to release phosphate was examined in deionized and natural waters under UV and sunlight irradiation using glyphosate as the organic phosphorus model. The results showed that the concentration of glyphosate decreased with irradiation time in the Fe3+/UV and Fe3+/sunlight systems and TOC gradually decreased, which confirmed that glyphosate was degraded by Fe3+. The amount of phosphate released from the photo-decomposition of glyphosate was higher in the presence of Fe3+ than that of the control experiment under UV and sunlight irradiation conditions, and the generation rate of phosphate also increased with increasing Fe3+concentrations. The formation of hydroxyl radicals (·OH) in the Fe3+/UV and Fe3+/sunlight systems was identified according to the photoluminescence spectra (PL) using coumarin as the trapping molecule, and the steady-state concentrations of ·OH for the Fe3+/UV and Fe3+/sunlight systems were 1.06 × 10−14 M and 0.09 × 10−14 M, respectively. When natural water was spiked with glyphosate and Fe3+, the phosphate that was released in the Fe3+ was higher than that of the control, and the phosphate that was released was inhibited when isopropanol was added to the reaction. All of these results demonstrate that the photochemical activity of Fe3+ has significantly impact in the release of phosphate from the photo-decomposition of organic phosphorus.

Phosphate solubilization in vitro by isolated Aspergillus niger and Aspergillus carbonarius

Two strains of Aspergillus, A. niger and A. carbonarius, were isolated from agricultural soil and a lake, respectively, in China. The two isolates could effectively release soluble phosphate in NBRIP medium containing Ca3(PO4)2 as the sole phosphorus source. Acidification of the broth seemed to be the major mechanism for phosphate solubilization by the isolates, and this had a significant correlation with a drop in pH. High-pressure liquid chromatography analysis indicated the participation of various organic acids, with gluconic acid as the principal component in the process of acidification in the broth. The isolates displayed trends of soluble phosphate release that closely matched the substantial increases in gluconic acid concentration. Abiotic incubation study using organic or inorganic acid to solubilize Ca3(PO4)2 indicated that the content of soluble phosphate released was significantly lower than that of the broth inoculated with the isolates. Higher release of soluble phosphate and pH reduction have occurred when ammonium rather than nitrate served as the sole source of nitrogen with Ca3(PO4)2.

Solubilization of aluminum phosphate by specific Penicillium spp.

The solubilization of hardly soluble aluminum phosphate (AlPO4) by specific Penicillium spp. isolated from wheat rhizospheric soils was investigated in Pikovskaya agar and liquid medium, respectively. Most of the Penicillium isolates except P. simplicissimum AP11 and P. variabile AP15 developed clear transparent zone around the colony margin in plate assays. Results of broth assays show that the Penicillium isolates can efficiently solubilize aluminum phosphate in Pikovskaya liquid medium, and vary in their capabilities to release soluble phosphate from aluminum phosphate. All the isolates exhibit different abilities to lower the pH and increase the titratable acidity in the broth compared to the control. HPLC analysis shows that most of the isolates except the species of P. aurantiogriseum can excrete different concentrations of organic acids, including gluconic acid, citric acid, oxalic acid, malic acid and tartaric acids, in the broth. The release of soluble phosphate by the isolate P. oxalicum AP2, which is the best solubilizer of aluminum phosphate among the isolates, is accompanied by a significant drop of pH and an obvious rise of titratable acidity during 7 d of aluminum phosphate-solubilizing experiments. The effects of temperature, initial pH, concentration of aluminum phosphate and shaking speed on aluminum phosphate solubilization by P. oxalicum AP2 were also investigated, and the maximum contents of soluble phosphate released are recorded at temperature 30 °C, initial pH 6, aluminum phosphate concentration 20 g/L, and shaking speed 160 r/min.

Effects of Acidiphilium cryptum on biosolubilization of rock phosphate in the presence of Acidithiobacillus ferrooxidans

The bioleaching of pyrite and biosolubilization of rock phosphate (RP) in 9K basal salts medium were compared by the following strains of an autotrophic acidophilic bacterium, Acidithiobacillus ferrooxidans, a heterotrophic acidophilic bacterium, Acidiphilium cryptum, and mixed culture of At. ferrooxidans and A. cryptum. The results show that A. cryptum is effective in enhancing the bioleaching of pyrite and biosolubilization of RP in the presence of At. ferrooxidans, although it could not oxidize pyrite and solubilize RP by itself. This effect is demonstrated experimentally that A. cryptum enhances a decrease in pH and an increase in redox potential, concentration of total soluble iron and planktonic part bacterial number in the broth during pyrite bioleaching processes by At. ferrooxidans. The mixed culture of At. ferrooxidans and A. cryptum leads to the most extensive soluble phosphate released at 30 °C. Pulp density exceeding 3% is shown to adversely influence the release of soluble phosphate by the consortium of At. ferrooxidans and A. cryptum. It is essential to add pyrite to the 9K basal salts medium for the biosolubilization of RP by the mixed culture of At. ferrooxidans and A. cryptum, and the percentage of soluble phosphate released is the greatest when the mass ratio of RP to pyrite is 1:2 or 1:3.

Yeast Pif1 Helicase Exhibits a One-base-pair Stepping Mechanism for Unwinding Duplex DNA

Kinetic analysis of the DNA unwinding and translocation activities of helicases is necessary for characterization of the biochemical mechanism(s) for this class of enzymes. Saccharomyces cerevisiae Pif1 helicase was characterized using presteady state kinetics to determine rates of DNA unwinding, displacement of streptavidin from biotinylated DNA, translocation on single-stranded DNA (ssDNA), and ATP hydrolysis activities. Unwinding of substrates containing varying duplex lengths was fit globally to a model for stepwise unwinding and resulted in an unwinding rate of ∼75 bp/s and a kinetic step size of 1 base pair. Pif1 is capable of displacing streptavidin from biotinylated oligonucleotides with a linear increase in the rates as the length of the oligonucleotides increased. The rate of translocation on ssDNA was determined by measuring dissociation from varying lengths of ssDNA and is essentially the same as the rate of unwinding of dsDNA, making Pif1 an active helicase. The ATPase activity of Pif1 on ssDNA was determined using fluorescently labeled phosphate-binding protein to measure the rate of phosphate release. The quantity of phosphate released corresponds to a chemical efficiency of 0.84 ATP/nucleotides translocated. Hence, when all of the kinetic data are considered, Pif1 appears to move along DNA in single nucleotide or base pair steps, powered by hydrolysis of 1 molecule of ATP.

Synaptic Released Zinc Promotes Tau Hyperphosphorylation by Inhibition of Protein Phosphatase 2A (PP2A)

Hyperphosphorylated tau is the major component of neurofibrillary tangles in Alzheimer disease (AD), and the tangle distribution largely overlaps with zinc-containing glutamatergic neurons, suggesting that zinc released in synaptic terminals may play a role in tau phosphorylation. To explore this possibility, we treated cultured hippocampal slices or primary neurons with glutamate or Bic/4-AP to increase the synaptic activity with or without pretreatment of zinc chelators, and then detected the phosphorylation levels of tau. We found that glutamate or Bic/4-AP treatment caused tau hyperphosphorylation at multiple AD-related sites, including Ser-396, Ser-404, Thr-231, and Thr-205, while application of intracellular or extracellular zinc chelators, or blockade of zinc release by extracellular calcium omission almost abolished the synaptic activity-associated tau hyperphosphorylation. The zinc release and translocation of excitatory synapses in the hippocampus were detected, and zinc-induced tau hyperphosphorylation was also observed in cultured brain slices incubated with exogenously supplemented zinc. Tau hyperphosphorylation induced by synaptic activity was strongly associated with inactivation of protein phosphatase 2A (PP2A), and this inactivation can be reversed by pretreatment of zinc chelator. Together, these results suggest that synaptically released zinc promotes tau hyperphosphorylation through PP2A inhibition.

Phosphate geochemistry, mineralization processes, and Thioploca distribution in shelf sediments off central Chile

Sediments underlying the major costal upwelling systems of the world oceans are hot-spots of modern formation of hydroxyapatites, often associated with benthic communities of large, nitrate-accumulating sulfur bacteria. We studied the association between phosphate release, organic phosphorus mineralization, and occurrence of dense communities of the filamentous sulfur bacteria, Thioploca spp., on the continental shelf off central Chile during the austral summer when high phytoplankton productivity and anoxic bottom water prevailed. Freshly deposited phytodetritus stimulated extremely high sulfate reduction rates, which supported a large Thioploca community of up to 100 g biomass per m 2. Effective bacterial sulfide uptake kept the sulfide concentration low, which enabled the accumulation of free iron, thus demonstrating intensive iron reduction concurrent with sulfate reduction. Phosphate released to the pore water reached 100–300 μM peak concentrations within the uppermost 0–5 cm and phosphate was lost to the overlying anoxic water column. The large phosphate release was not directly related to the presence of Thioploca but was rather the result of a high deposition and mineralization rate of fresh organic detritus. Although the pore water was super–saturated with respect to hydroxyapatite, this mineral was only a minor P-component in the sediment. Most solid-phase phosphate was bound to iron.

ATP-dependent MurE ligase in Mycobacterium tuberculosis: Biochemical and structural characterisation

New therapies are required against Mycobacterium tuberculosis and its cell wall peptidoglycan biosynthesis is a potential therapeutic target. UDP-MurNAc-tripeptide ligase (MurE) is a member of the ATP-dependent ligase family, which incorporate amino acids including meso-diaminopimelic acid (m-DAP) into peptidoglycan during synthesis in a species-specific manner. In the present study, we have cloned, over-expressed, and characterised MurE from M. tuberculosis (Mtb-MurE). The crystal structure has been determined at 3.0A resolution in the presence of the substrate UDP-MurNAc-l-Ala-d-Glu (UAG). The activity of the enzyme was measured through estimating inorganic phosphate released in a non-radioactive high-throughput colourimetric assay. UDP-MurNAc-l-Ala-d-Glu-m-DAP (UMT) formation coupled to inorganic phosphate release was confirmed by HPLC and mass spectrometric analyses. Kinetic constants were determined for a range of natural substrates using optimised conditions. From our findings, it is evident that Mtb-MurE is highly specific in adding m-DAP to UDP-MurNAc-dipeptide and ATP-hydrolysis is an absolute requirement for its activity.

Adenine Nucleotide-dependent Regulation of Assembly of Bacterial Tubulin-like FtsZ by a Hypermorph of Bacterial Actin-like FtsA

Cytokinesis in bacteria depends upon the contractile Z ring, which is composed of dynamic polymers of the tubulin homolog FtsZ as well as other membrane-associated proteins such as FtsA, a homolog of actin that is required for membrane attachment of the Z ring and its subsequent constriction. Here we show that a previously characterized hypermorphic mutant FtsA (FtsA*) partially disassembled FtsZ polymers in vitro. This effect was strictly dependent on ATP or ADP binding to FtsA* and occurred at substoichiometric levels relative to FtsZ, similar to cellular levels. Nucleotide-bound FtsA* did not affect FtsZ GTPase activity or the critical concentration for FtsZ assembly but was able to disassemble preformed FtsZ polymers, suggesting that FtsA* acts on FtsZ polymers. Microscopic examination of the inhibited FtsZ polymers revealed a transition from long, straight polymers and polymer bundles to mainly short, curved protofilaments. These results indicate that a bacterial actin, when activated by adenine nucleotides, can modify the length distribution of bacterial tubulin polymers, analogous to the effects of actin-depolymerizing factor/cofilin on F-actin.