Phosphate Biomineralization Research Articles

Mechanistic insights into tris(2-chloroisopropyl) phosphate biomineralization coupled with lead (II) biostabilization driven by denitrifying bacteria

The ubiquity and persistence of organophosphate esters (OPEs) and heavy metal (HMs) pose global environmental risks. This study explored tris(2-chloroisopropyl)phosphate (TCPP) biomineralization coupled to lead (Pb2+) biostabilization driven by denitrifying bacteria (DNB). The domesticated DNB achieved synergistic bioremoval of TCPP and Pb2+ in the batch bioreactor (efficiency: 98 %).TCPP mineralized into PO43− and Cl−, and Pb2+ precipitated with PO43−. The TCPP-degrading/Pb2+-resistant DNB: Achromobacter, Pseudomonas, Citrobacter, and Stenotrophomonas, dominated the bacterial community, and synergized TCPP biomineralization and Pb2+ biostabilization. Metagenomics and metaproteomics revealed TCPP underwent dechlorination, hydrolysis, the TCA cycle-based dissimilation, and assimilation; Pb2+ was detoxified via bioprecipitation, bacterial membrane biosorption, EPS biocomplexation, and efflux out of cells. TCPP, as an initial donor, along with NO3−, as the terminal acceptor, formed a respiratory redox as the primary energy metabolism. Both TCPP and Pb2+ can stimulate phosphatase expression, which established the mutual enhancements between their bioconversions by catalyzing TCPP dephosphorylation and facilitating Pb2+ bioprecipitation. TCPP may alleviate the Pb2+-induced oxidative stress by aiding protein phosphorylation. 80 % of Pb2+ converted into crystalized pyromorphite. These results provide the mechanistic foundations and help develop greener strategies for synergistic bioremediation of OPEs and HMs.

Fluorochemicals from fluorspar via a phosphate-enabled mechanochemical process that bypasses HF.

All fluorochemicals-including elemental fluorine and nucleophilic, electrophilic, and radical fluorinating reagents-are prepared from hydrogen fluoride (HF). This highly toxic and corrosive gas is produced by the reaction of acid-grade fluorspar (>97% CaF2) with sulfuric acid under harsh conditions. The use of fluorspar to produce fluorochemicals via a process that bypasses HF is highly desirable but remains an unsolved problem because of the prohibitive insolubility of CaF2. Inspired by calcium phosphate biomineralization, we herein disclose a protocol of treating acid-grade fluorspar with dipotassium hydrogen phosphate (K2HPO4) under mechanochemical conditions. The process affords a solid composed of crystalline K3(HPO4)F and K2-xCay(PO3F)a(PO4)b, which is found suitable for forging sulfur-fluorine and carbon-fluorine bonds.

Extracellular biomineralization of uranium and its toxicity alleviation to Bacillus thuringiensis X-27

Uranium biomineralization can slow uranium migration in the environment and thus prevent it from further contaminating the surroundings. Investigations into the uranium species, pH, inorganic phosphate (Pi) concentration, and microbial viability during biomineralization by microorganisms are crucial for understanding the mineralization mechanism. In this study, Bacillus thuringiensis X-27 was isolated from soil contaminated with uranium and was used to investigate the formation process of uranium biominerals induced by X-27. The results showed that as biomineralization proceeded, amorphous uranium-containing deposits were generated and transformed into crystalline minerals outside cells, increasing the overall concentration of uramphite. This is a cumulative rather than abrupt process. Notably, B. thuringiensis X-27 precipitated uranium outside the cell surface within 0.5 h, while the release of Pi into the extracellular environment and the change of pH to alkalescence further promoted the formation of uramphite. In addition, cell viability determination showed that the U(VI) biomineralization induced by B. thuringiensis X-27 was instrumental in alleviating the toxicity of U(VI) to cells. This work offers insight into the mechanism of U(VI) phosphate biomineralization and is a reference for bioremediation-related studies.

Effect of different phosphate sources on uranium biomineralization by the Microbacterium sp. Be9 strain: A multidisciplinary approach study.

Industrial activities related with the uranium industry are known to generate hazardous waste which must be managed adequately. Amongst the remediation activities available, eco-friendly strategies based on microbial activity have been investigated in depth in the last decades and biomineralization-based methods, mediated by microbial enzymes (e.g., phosphatase), have been proposed as a promising approach. However, the presence of different forms of phosphates in these environments plays a complicated role which must be thoroughly unraveled to optimize results when applying this remediation process. In this study, we have looked at the effect of different phosphate sources on the uranium (U) biomineralization process mediated by Microbacterium sp. Be9, a bacterial strain previously isolated from U mill tailings. We applied a multidisciplinary approach (cell surface characterization, phosphatase activity, inorganic phosphate release, cell viability, microscopy, etc.). It was clear that the U removal ability and related U interaction mechanisms by the strain depend on the type of phosphate substrate. In the absence of exogenous phosphate substrate, the cells interact with U through U phosphate biomineralization with a 98% removal of U within the first 48 h. However, the U solubilization process was the main U interaction mechanism of the cells in the presence of inorganic phosphate, demonstrating the phosphate solubilizing potential of the strain. These findings show the biotechnological use of this strain in the bioremediation of U as a function of phosphate substrate: U biomineralization (in a phosphate free system) and indirectly through the solubilization of orthophosphate from phosphate (P) containing waste products needed for U precipitation.

Tunable Enzyme-Assisted Mineralization of Apatitic Calcium Phosphate by Homogeneous Catalysis

While it has long been mimicked by simple precipitation reactions under biologically relevant conditions, calcium phosphate biomineralization is a complex process, which is highly regulated by physicochemical factors and involves a variety of proteins and other biomolecules. Alkaline phosphatase (ALP), in particular, is a conductor of sorts, directly regulating the amount of orthophosphate ions available for mineralization. Herein, we explore enzyme-assisted mineralization in the homogeneous phase as a method for biomimetic mineralization and focus on how relevant ionic substitution types affect the obtained minerals. For this purpose, mineralization is performed over a range of enzyme substrate concentrations and fluoride concentrations at physiologically relevant conditions (pH 7.4, T = 37 °C). Refinement of X-ray diffraction data is used to study the crystallographic unit cell parameters for evidence of ionic substitution in the lattice, and infrared (IR) spectroscopy and X-ray photoelectron spectroscopy (XPS) are used for complementary information regarding the chemical composition of the minerals. The results show the formation of substituted hydroxyapatite (HAP) after 48 h mineralization in all conditions. Interestingly, an expansion of the crystalline unit cell with an increasing concentration of the enzyme substrate is observed, with only slight changes in the particle morphology. On the contrary, by increasing the amount of fluoride, while keeping the enzyme substrate concentration unchanged, a contraction of the crystalline unit cell and the formation of elongated, well-crystallized rods are observed. Complementary IR and XPS data indicate that these trends are explained by the incorporation of substituted ions, namely CO32- and F-, in the HAP lattice at different positions.

In Situ Fluorescence Probing of the Formation of Calcium Phosphate Prenucleation Clusters.

Initial-stage prenucleation clusters (PNCs) are critical in calcium phosphate (CaP) biomineralization and thus the formation mechanisms of human bones and teeth. However, several features of PNCs require further examination, e.g., structure, ionic stoichiometry, kinetics, thermodynamics, and nucleation mechanism. In this study, we used poly(acrylic acid) (PAA)-Ca(Eu) complexes with partial Eu3+ substitution as pre-PNCs and established a fluorescence method to study PNC formation in situ based on Eu-O charge-transfer transitions. The kinetics and thermodynamics of PNC formation were explored by probing the fluorescence changes of Eu-O charge-transfer transitions during bonding between the pre-PNCs and PO43-. PNC formation was consistent with the pseudo-second-order kinetic and Langmuir isothermal adsorption models. The flexible structures of PNCs aided in regulating the subsequent nucleation and crystallization. This study provides an in situ fluorescence probing method with critical guiding significance in addressing the features of PNC formation, in addition to biomineralization.

Vertebrate Taphonomy and Diagenesis: Implications of Structural and Compositional Alterations of Phosphate Biominerals

Biominerals are recorders of evolution and palaeoenvironments. Predation is one of the most frequent modes leading to the concentration of small vertebrates in fossil assemblages. Consumption by predators produces damages on bones and teeth from prey species, and one of the greatest challenges to taphonomists is differentiating original biological and secondary, geologically altered attributes of fossils. Excellent morphological preservation is often used to assume that the structure and composition of fossils are not modified. Nevertheless, during predation and fossilization, both the physical structure and chemical composition of enamel, dentine and bone are altered, the degree and extent of which varies from site to site, depending on the nature of the burial environment. A relationship between the surficial alterations and the compositional changes which take place during fossilization has yet to be established. Herein, I present a review of old and recent taphonomic studies that collectively reveal the wide diversity of microstructural and chemical changes that typically take place during fossilization of vertebrate remains, including common taphonomic biases and the challenges inherent to reconstructing the history of vertebrate fossil assemblages.

Effect of Temperature and Cell Viability on Uranium Biomineralization by the Uranium Mine Isolate Penicillium simplicissimum.

The remediation of heavy-metal-contaminated sites represents a serious environmental problem worldwide. Currently, cost- and time-intensive chemical treatments are usually performed. Bioremediation by heavy-metal-tolerant microorganisms is considered a more eco-friendly and comparatively cheap alternative. The fungus Penicillium simplicissimum KS1, isolated from the flooding water of a former uranium (U) mine in Germany, shows promising U bioremediation potential mainly through biomineralization. The adaption of P. simplicissimum KS1 to heavy-metal-contaminated sites is indicated by an increased U removal capacity of up to 550 mg U per g dry biomass, compared to the non-heavy-metal-exposed P. simplicissimum reference strain DSM 62867 (200 mg U per g dry biomass). In addition, the effect of temperature and cell viability of P. simplicissimum KS1 on U biomineralization was investigated. While viable cells at 30°C removed U mainly extracellularly via metabolism-dependent biomineralization, a decrease in temperature to 4°C or use of dead-autoclaved cells at 30°C revealed increased occurrence of passive biosorption and bioaccumulation, as confirmed by scanning transmission electron microscopy. The precipitated U species were assigned to uranyl phosphates with a structure similar to that of autunite, via cryo-time-resolved laser fluorescence spectroscopy. The major involvement of phosphates in U precipitation by P. simplicissimum KS1 was additionally supported by the observation of increased phosphatase activity for viable cells at 30°C. Furthermore, viable cells actively secreted small molecules, most likely phosphorylated amino acids, which interacted with U in the supernatant and were not detected in experiments with dead-autoclaved cells. Our study provides new insights into the influence of temperature and cell viability on U phosphate biomineralization by fungi, and furthermore highlight the potential use of P. simplicissimum KS1 particularly for U bioremediation purposes. Graphical

Enzymatic Approach in Calcium Phosphate Biomineralization: A Contribution to Reconcile the Physicochemical with the Physiological View.

Biomineralization is the process by which organisms produce hard inorganic matter from soft tissues with outstanding control of mineral deposition in time and space. For this purpose, organisms deploy a sophisticated “toolkit” that has resulted in significant evolutionary innovations, for which calcium phosphate (CaP) is the biomineral selected for the skeleton of vertebrates. While CaP mineral formation in aqueous media can be investigated by studying thermodynamics and kinetics of phase transitions in supersaturated solutions, biogenic mineralization requires coping with the inherent complexity of biological systems. This mainly includes compartmentalization and homeostatic processes used by organisms to regulate key physiological factors, including temperature, pH and ion concentration. A detailed analysis of the literature shows the emergence of two main views describing the mechanism of CaP biomineralization. The first one, more dedicated to the study of in vivo systems and supported by researchers in physiology, often involves matrix vesicles (MVs). The second one, more investigated by the physicochemistry community, involves collagen intrafibrillar mineralization particularly through in vitro acellular models. Herein, we show that there is an obvious need in the biological systems to control both where and when the mineral forms through an in-depth survey of the mechanism of CaP mineralization. This necessity could gather both communities of physiologists and physicochemists under a common interest for an enzymatic approach to better describe CaP biomineralization. Both homogeneous and heterogeneous enzymatic catalyses are conceivable for these systems, and a few preliminary promising results on CaP mineralization for both types of enzymatic catalysis are reported in this work. Through them, we aim to describe the relevance of our point of view and the likely findings that could be obtained when adding an enzymatic approach to the already rich and creative research field dealing with CaP mineralization. This complementary approach could lead to a better understanding of the biomineralization mechanism and inspire the biomimetic design of new materials.

A Novel Magnetotactic Alphaproteobacterium Producing Intracellular Magnetite and Calcium-Bearing Minerals.

Magnetotactic bacteria (MTB) are prokaryotes that form intracellular magnetite (Fe3O4) or greigite (Fe3S4) nanocrystals with tailored sizes, often in chain configurations. Such magnetic particles are each surrounded by a lipid bilayer membrane, called a magnetosome, and provide a model system for studying the formation and function of specialized internal structures in prokaryotes. Using fluorescence-coupled scanning electron microscopy, we identified a novel magnetotactic spirillum, XQGS-1, from freshwater Xingqinggong Lake, Xi'an City, Shaanxi Province, China. Phylogenetic analyses based on 16S rRNA gene sequences indicate that strain XQGS-1 represents a novel genus of the Alphaproteobacteria class in the Proteobacteria phylum. Transmission electron microscopy analyses reveal that strain XQGS-1 forms on average 17 ± 3 magnetite magnetosome particles with an ideal truncated octahedral morphology, with an average length and width of 88.3 ± 11.7 nm and 83.3 ± 11.0 nm, respectively. They are tightly organized into a single chain along the cell long axis close to the concave side of the cell. Intrachain magnetic interactions likely result in these large equidimensional magnetite crystals behaving as magnetically stable single-domain particles that enable bacterial magnetotaxis. Combined structural and chemical analyses demonstrate that XQGS-1 cells also biomineralize intracellular amorphous calcium phosphate (2 to 3 granules per cell; 90.5- ± 19.3-nm average size) and weakly crystalline calcium carbonate (2 to 3 granules per cell; 100.4- ± 21.4-nm average size) in addition to magnetite. Our results expand the taxonomic diversity of MTB and provide evidence for intracellular calcium phosphate biomineralization in MTB. IMPORTANCE Biomineralization is a widespread process in eukaryotes that form shells, teeth, or bones. It also occurs commonly in prokaryotes, resulting in more than 60 known minerals formed by different bacteria under wide-ranging conditions. Among them, magnetotactic bacteria (MTB) are remarkable because they might represent the earliest organisms that biomineralize intracellular magnetic iron minerals (i.e., magnetite [Fe3O4] or greigite [Fe3S4]). Here, we report a novel magnetotactic spirillum (XQGS-1) that is phylogenetically affiliated with the Alphaproteobacteria class. In addition to magnetite crystals, XQGS-1 cells form intracellular submicrometer calcium carbonate and calcium phosphate granules. This finding supports the view that MTB are also an important microbial group for intracellular calcium carbonate and calcium phosphate biomineralization.

High-efficient microbial immobilization of solved U(VI) by the Stenotrophomonas strain Br8

The environmental impact of uranium released during nuclear power production and related mining activity is an issue of great concern. Innovative environmental-friendly water remediation strategies, like those based on U biomineralization through phosphatase activity, are desirable. Here, we report the great U biomineralization potential of Stenotrophomonas sp. Br8 CECT 9810 over a wide range of physicochemical and biological conditions. Br8 cells exhibited high phosphatase activity which mediated the release of orthophosphate in the presence of glycerol-2-phosphate around pH 6.3. Mobile uranyl ions were bioprecipitated as needle-like fibrils at the cell surface and in the extracellular space, as observed by Scanning Transmission Electron Microscopy (STEM). Extended X-Ray Absorption Fine Structure (EXAFS) and X-Ray Diffraction (XRD) analyses showed the local structure of biogenic U precipitates to be similar to that of meta-autunite. In addition to the active U phosphate biomineralization process, the cells interact with this radionuclide through passive biosorption, removing up to 373 mg of U per g of bacterial dry biomass. The high U biomineralization capacity of the studied strain was also observed under different conditions of pH, temperature, etc. Results presented in this work will help to design efficient U bioremediation strategies for real polluted waters.

Simulation of Calcium Phosphate Prenucleation Clusters in Aqueous Solution: Association beyond Ion Pairing.

Classical molecular dynamics simulations and free energy methods have been used to obtain a better understanding of the molecular processes occurring prior to the first nucleation event for calcium phosphate biominerals. The association constants for the formation of negatively charged complexes containing calcium and phosphate ions in aqueous solution have been computed, and these results suggest that the previously proposed calcium phosphate building unit, [Ca(HPO4)3]4–, should only be present in small amounts under normal experimental conditions. However, the presence of an activation barrier for the removal of an HPO42– ion from this complex indicates that this species could be kinetically trapped. Aggregation pathways involving CaHPO4, [Ca(HPO4)2]2–, and [Ca(HPO4)3]4– complexes have been explored with the finding that dimerization is favorable up to a Ca/HPO4 ratio of 1:2.

Unstructured Proteins in Biological Structures: The Case of Human Teeth from a Protein Chemist's Perspective

Proteins are the biomacromolecules that work downstream to genes in every living system. It is these protein molecules that are responsible for a plethora of functions in our body, ranging from building skeleton tissues to transmitting signal from one place to others. There was a period when biologists have the view that for a protein to function, the structure is of supreme importance. Nevertheless, this structure‐function paradigm is fairly obeyed by most of the proteins, if not all of them. Last two decades have witnessed the increased reports of proteins that do not have a stable three‐dimensional structure but are very much indispensable for the cellular and biological activities. These unstructured proteins are now known as “Intrinsically disordered proteins” (IDPs). Interestingly, these IDPs function not only in isolation (e.g. transcription factors) but helps in the formation of hard tissues like teeth, bone, and mollusks shell, by governing a process called biomineralization. Biomineralization is the process by which living organisms produce minerals, often to harden or stiffen existing tissues. Such tissues are called mineralized tissues.We have isolated total protein extracts from a healthy molar, premolar, canine and incisor teeth. We studied their protein secondary structural characteristics using various biophysical techniques. We have also investigated the effects of these protein extracts on mineralization of calcium phosphate through in vitro biomineralization assay followed by size‐based measurement analysis via Nanoparticle tracking analysis (NTA). The biophysical studies confirmed the presence of IDPs in the majority within these protein extracts. Moreover, these extracts were shown to elevate calcium phosphate biomineralization in a time‐dependent manner. This is the first time the effect of the whole protein extract was taken into account which in our view more closely imitates the in vivo process of tooth biomineralization. Furthermore, we also evaluated the differential proteome profile of the individual type of human teeth through Mass spectroscopy‐based method. The comparative qualitative and quantitative data of proteins from these different human teeth were obtained. The implication of these results will be discussed.Support or Funding InformationThere was no support or funding from any organizationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Bioremediation of strontium and technetium contaminated groundwater using glycerol phosphate

Groundwater at legacy nuclear facilities around the world is contaminated with radionuclides including strontium-90 and technetium-99, which are often present as co-contaminants. Here we investigated whether biostimulation of indigenous microbial communities by glycerol phosphate can co-treat 90Sr through incorporation into phosphate biominerals, and 99Tc through microbially-induced reduction of the sediment to form less mobile Tc(IV) phases via reaction with reduced species (e.g. Fe(II)). Results showed that 95% of Sr was removed from solution in sediment microcosms treated with glycerol phosphate, and sequential extraction showed that ~18% of the Sr in the resulting solid phase was associated with the pH 5 Na-acetate fraction and 75% was in the ion exchangeable fraction. This removal and partitioning to recalcitrant phases during glycerol phosphate treatment was greater than in the untreated controls, where only 60% of Sr was removed from solution, and of the solid-associated Sr, 95% was present in the exchangeable fraction. Fitting of Sr K-edge EXAFS spectra confirmed these findings, with shell by shell fitting suggesting ~30% of sediment-associated Sr was present in a coordination environment consistent with phosphate biominerals following glycerol phosphate treatment, whilst Sr was present only as outer-sphere complexes in the controls. In addition,16S rRNA sequencing of sediments stimulated with glycerol phosphate demonstrated the growth of potential phosphate-solubilising species such as Chryseobacterium and Serratia spp. Finally, glycerol phosphate treatment stimulated bioreduction via addition of electron donor in the form of glycerol to the system, in turn this stimulated the removal of 99Tc from solution concomitant with microbial Fe(III) reduction to form poorly soluble hydrous Tc(IV)O2 like phases. In sediments amended with an electron donor, the microbial community also reflected the onset of bioreduction with an increased relative abundance of Fe(III) and sulfate-reducing bacteria such as Geothrix, Geobacter and Desulfobulbus spp. Overall these results suggest application of glycerol phosphate offers a promising bioremediation strategy to co-treat both 90Sr and 99Tc contaminated groundwaters, and promotes the formation of Sr-phosphate and Tc(IV) bearing biominerals when reducing conditions are maintained. Combined with past work which shows the scavenging of uranium from solution following addition of glycerol phosphate, this extends the scope for glycerol phosphate as a treatment for radioactive contamination in groundwaters.

Biomineralization of calcium phosphate revealed by in situ liquid-phase electron microscopy

Calcium phosphate biomineralization is essential to the formation of bones and teeth, and other pathological calcifications. Unravelling the mechanism of calcium phosphate nucleation and growth contributes significantly to understanding diseases caused by pathological mineralization, and also to designing biomimetic materials with suitable properties. Recently, calcium phosphate was proposed to mineralize following a non-classical crystal growth pathway of pre-nucleation cluster aggregation. Liquid-phase transmission electron microscopy allows dynamic processes to be recorded continuously inside liquid. Here we present direct evidence, based on continuous monitoring in liquid, to confirm that calcium phosphate mineralization from simulated body fluid occurs by particle attachment, shown with nanoscale spatial resolution and sufficient temporal resolution. This work may lay the foundation for future investigation of mineralization in other relevant biological systems in humans and vertebrates.

Mechanisms of Modulation of Calcium Phosphate Pathological Mineralization by Mobile and Immobile Small-Molecule Inhibitors.

Potential pathways for inhibiting crystal growth are via either disrupting local microenvironments surrounding crystal-solution interfaces or physically blocking solute molecule attachment. However, the actual mode of inhibition may be more complicated due to the characteristic time scale for the inhibitor adsorption and relaxation to a well-bound state at crystal surfaces. Here we demonstrate the role of citrate (CA) and hydroxycitrate (HCA) in brushite (DCPD, CaHPO4·2H2O) crystallization over a broad range of both inhibitor concentrations and supersaturations by in situ atomic force microscopy (AFM). We observed that both inhibitors exhibit two distinct actions: control of surface crystallization by the decrease of step density at high supersaturations and the decrease of the [1̅00]Cc step velocity at high inhibitor concentration and low supersaturation. The switching of the two distinct modes depends on the terrace lifetime, and the slow kinetics along the [1̅00]Cc step direction provides specific sites for the newly formed dislocations. Molecular modeling shows the strong HCA-crystal interaction by molecular recognition, explaining the AFM observations for the formation of new steps and surface dissolution along the [101]Cc direction due to the introduction of strong localized strain in the crystal lattice. These direct observations highlight the importance of the inhibitor coverage on mineral surfaces, as well as the solution supersaturation in predicting the inhibition efficacy, and reveal an improved understanding of inhibition of calcium phosphate biomineralization, with clinical implications for the full therapeutic potential of small-molecule inhibitors for kidney stone disease.

Uranium biomineralization induced by a metal tolerant Serratia strain under acid, alkaline and irradiated conditions.

It has become increasingly apparent that the environmental microorganisms residing in uranium (U) enriched sites offer the possibility of understanding the biological mechanisms catalyzing the processes important for uranium bioremediation. Here, we present the results of uranium biomineralization over a wide pH range by a metal tolerant Serratia sp. strain OT II 7 isolated from the subsurface soil of a U ore deposit at Domiasiat in India. The Serratia cells actively expressed acid and alkaline phosphatase enzymes which hydrolyzed differential amounts of phosphate from an organophosphate substrate in the presence of uranium between pH 5 to 9. These cells precipitated ∼91% uranium from aqueous solutions supplemented with 1 mM uranyl nitrate at pH 5 within 120 h. More rapid precipitation was observed at pH 7 and 9 wherein the cells removed ∼93-94% of uranium from solutions containing 1 mM uranyl carbonate within 24 h. The aqueous uranyl speciation prevalent under the studied pH conditions influenced the localization of crystalline uranyl phosphate precipitates, which in turn, impacted the cell viability to a great extent. Furthermore, the cells tolerated up to ∼1.6 kGy 60Co gamma radiation and their uranium precipitation abilities at pH 5, 7 and 9 were uncompromised even after exposure to a high dose of ionizing radiation. Overall, this study establishes the ecological adaptation of a natural strain like Serratia in a uranium enriched environment and corroborates its contribution towards uranium immobilization in contaminated subsurfaces through the formation of stable uranyl phosphate minerals over a wide pH range.

Calcium phosphate in plant trichomes: the overlooked biomineral.

Calcium phosphate was unknown as a plant biomineral until recently reported in Neotropical Loasaceae. Here, we demonstrate its widespread occurrence in the trichomes of several plant families, including Brassicaceae. Calcium phosphate is the primary biomineral in, e.g., the bones and teeth of higher animals; in plants, it was only recently discovered in the stinging hairs and scabrid-glochidiate trichomes of South American Loasaceae (Ensikat et al. in Sci Rep UK 6:26073, 2016), where it appears to be deposited highly specifically, often replacing the common plant biomineral silica. We initiated a broader survey in a range of different plant orders to investigate a possibly wider distribution of calcium phosphate biomineralization in plants. Scanning electron microscopy with EDX element analysis and mapping was used for the detection of the biominerals: calcium phosphate, calcium carbonate, and silica in the trichomes of several common plant species of different orders. Results were authenticated with Raman spectroscopy. Calcium phosphate was found in the trichomes of several species in the orders Malpighiales, Rosales, Boraginales, and Brassicales. It occurred in trichome tips, replacing the more common silica, or together with silica and calcium carbonate at specific locations in the trichome cell walls. Most surprisingly, it was found in the trichomes of Arabidopsis thaliana, one of the most studied plant species-where it had been overlooked so far. The wide distribution of calcium phosphate as plant biomineral here demonstrated and the striking mineralization patterns with three different biominerals in the walls of single-celled trichomes underscore an unexpected complexity in plant biomineralization.

Energetic Basis for Inhibition of Calcium Phosphate Biomineralization by Osteopontin.

Calcium oxalate kidney stones form attached to Randall's plaques (RP), calcium phosphate (Ca-P) deposits on the renal papillary surface. Osteopontin (OPN) suppresses crystal growth in the complex process of urinary stone formation, but the inhibitory role of active domains of OPN involved in the initial formation of the RPs attached to epithelial cells has yet to be clarified. Here we demonstrate the thermodynamic basis for how OPN sequences regulate the onset of Ca-P mineral formation on lipid rafts as a model membrane. We first quantify the kinetics of hydroxyapatite (HAP) nucleation on membrane substrates having liquid-condensed (LC) and liquid-expanded (LE) phases using in situ atomic force microscopy (AFM). We find that rates are sequence-dependent, and the thermodynamic barrier to nucleation is reduced by minimizing the interfacial free energy γ. Combined with single-molecule determination of the binding energy (ΔGB) of the OPN peptide segments adsorbed to the HAP (100) face, we show a linear relationship of γ and ΔGB, suggesting that the increase in the nucleation barriers correlates with strong peptide-crystal nuclei binding. These findings reveal fundamental energetic clues for inhibition of membrane-mediated nucleation by sequence motifs and subdomains within the OPN protein through spatial location of charged moieties and provide insight connecting peripheral cell membranes to pathological mineralization.

Fungal biomineralization of lead phosphates on the surface of lead metal

The present work describes lead phosphate biomineralization by a wood-decaying fungal isolate, Penicillium chrysogenum A15, after incubation in solid medium with metallic lead (Pb shot pellets). This fungal isolate showed high tolerance to this element, being able to grow on solid medium containing 8mMPb(II). Environmental Scanning Electron Microscope (ESEM) observations of abiotic controls showed the presence of lead oxide crystals on the lead shot surface after 8weeks incubation. However, in presence of P. chrysogenum, lead phosphate mineral phases were detected as aggregates on the surface of lead shots after 2-weeks and 8-weeks incubation. The morphology of the biotically deposited secondary minerals showed significant differences in comparison to those produced under abiotic conditions, appearing globular, prismatic and acicular crystals. High Resolution Transmission Electron Microscope (HRTEM) analysis indicated deposition of lead phosphates on the cell surface which could be considered as Pb resistance mechanism used by this isolate to cope with toxicity of lead. This study extends the spectrum of fungal isolates with potential on the biomineralization of lead phosphates useful for the bioremediation of lead contaminated sites.