Tobias Reich

Complexation of Uranium by Cells and S-Layer Sheets of Bacillus sphaericus JG-A12

ABSTRACT Bacillus sphaericus JG-A12 is a natural isolate recovered from a uranium mining waste pile near the town of Johanngeorgenstadt in Saxony, Germany. The cells of this strain are enveloped by a highly ordered crystalline proteinaceous surface layer (S-layer) possessing an ability to bind uranium and other heavy metals. Purified and recrystallized S-layer proteins were shown to be phosphorylated by phosphoprotein-specific staining, inductive coupled plasma mass spectrometry analysis, and a colorimetric method. We used extended X-ray absorption fine-structure (EXAFS) spectroscopy to determine the structural parameters of the uranium complexes formed by purified and recrystallized S-layer sheets of B. sphaericus JG-A12. In addition, we investigated the complexation of uranium by the vegetative bacterial cells. The EXAFS analysis demonstrated that in all samples studied, the U(VI) is coordinated to carboxyl groups in a bidentate fashion with an average distance between the U atom and the C atom of 2.88 ± 0.02 Å and to phosphate groups in a monodentate fashion with an average distance between the U atom and the P atom of 3.62 ± 0.02 Å. Transmission electron microscopy showed that the uranium accumulated by the cells of this strain is located in dense deposits at the cell surface.

Structure of uranium sorption complexes at montmorillonite edge sites

Summary Extended X-ray absorption fine structure (EXAFS) spectroscopy at the uranium LIII-edge was used for determining the structural environment of aqueous uranyl sorbed onto montmorillonite. The study reveals that uranyl uptake at pH ∼5-∼7 and at an initial uranyl concentration of 5×10−5 M takes place at amphoteric surface hydroxyl sites as inner-sphere complex. The measured bond distances between uranium and the equatorial oxygen atoms are in the range of 2.34 Å and 2.37 Å indicating an inner-sphere coordination. At ∼3.4 Å the presence of a U-Al backscattering pair was determined. This backscattering pair indicates that the binding of the uranyl unit to amphoteric surface hydroxyl sites occurs preferred as a bidentate inner-sphere complex on aluminol groups.

Interaction of uranium(VI) with various modified and unmodified natural and synthetic humic substances studied by EXAFS and FTIR spectroscopy

Abstract The complexation of uranium(VI) by humic acids (HAs) and fulvic acids (FAs) was studied to obtain information on the binding of uranium(VI) onto functional groups of humic substances. For this, various natural and synthetic HAs were chemically modified resulting in HAs with blocked phenolic OH groups. Both from the original and from the modified humic substances, solid uranyl humate complexes were prepared at pH 2. FTIR and extended X-ray absorption fine structure (EXAFS) spectroscopy were applied to study the chemical modification process of humic substances, to study the structure of uranyl humate complexes and to evaluate the effect of individual functional groups of humic substances (carboxylic and phenolic OH groups) on the complexation of uranyl ions. The results confirmed the predominant blocking of phenolic OH groups in the modified HAs. These modified HAs are suitable model substances to study the role of phenolic OH groups of HAs in dependence on pH. By EXAFS spectroscopy, identical structural parameters were determined for all uranyl humates. Axial UO bond distances of 1.78 A were determined. In the equatorial plane approximately five oxygen atoms were found at a mean distance of 2.39 A. The blocking of phenolic OH groups of HAs did not change the near-neighbor surrounding of uranium(VI) in uranyl humate complexes. Thus, the results confirmed that predominantly HA carboxylate groups are responsible for binding of uranyl ions and that the influence of phenolic OH groups is insignificant under the applied experimental conditions. The carboxylate groups are monodentate coordinated to uranyl ions.

Characterization of U(VI)-Acidithiobacillus ferrooxidans complexes using EXAFS, transmission electron microscopy, and energy-dispersive X-ray analysis

Summary We used a combination of Extended X-ray Absorption Fine Structure (EXAFS) spectroscopy, Transmission Electron Microscopy (TEM) and Energy-Dispersive X-ray (EDX) analysis to conduct molecular scale studies on U(VI) interaction with three recently described eco-types of Acidithiobacillus ferrooxidans. On the basis of the information obtained by using these methods, we concluded that uranyl phosphate complexes were formed by the cells of the three eco-types studied. The uranium accumulated by A. ferrooxidans cells was located mainly within the extracellular polysaccharides, and on the cell wall. Smaller amounts were also observed in the cytoplasm.

The colloid chemistry of acid rock drainage solution from an abandoned Zn–Pb–Ag mine

Abstract Acid rock drainage (ARD) solution from an abandoned ore mine (pH 2.7, SO2−4 concentration 411 mmol/l, Fe concentration 93.5 mmol/l) was investigated by photon correlation spectroscopy, centrifugation, filtration, ultrafiltration, scanning electron microscopy, ICP–MS, AAS, ion chromatography, TOC analysis, and extended X-ray absorption fine structure (EXAFS) spectroscopy. A colloid concentration of ⩾1 g/l was found. The prevailing particle size was

Uranium speciation in plants

Several spectroscopic methods specially the time-resolved laser-induced fluorescence spectroscopy and the X-ray-absorption spectroscopy were used for the determination of the uranium speciation in plants. Differences between the uranium speciation in the initial solution and inside the plants could be detected. The chemical speciation of uranium is identical in the roots, shoot axis and leaf. It is independent from the uranium speciation in the initial solution and the type of the plant. The results indicate that the uranium is predominantly bound as uranium phosphate (phosphoryl) groups in the plants.

Uranyl(VI) complexes with alpha-substituted carboxylic acids in aqueous solution

Summary The complex formation in the binary uranium(VI)-glycolate, -α-hydoxyisobutyrate, -α-aminoisobutyrate systems in 1.0 M NaClO4 medium was studied by means of UV-vis, TRLFS, and EXAFS. An increase in absorption and a red shift of the spectra, 5 nm compared to the free UO22+, indicate a complex formation between UO22+ and α-substituted carboxylic acids already at pH 2. 1:1 complexes dominate the uranyl speciation in the glycolate, α-hydoxyisobutyrate, and α-aminoisobutyrate system at pH 2 and 3, respectively. At higher ligand concentrations a 1:2 complex between UO22+ and α-aminoisobutyric acid was observed. There is a very strong quenching of the U(VI) fluorescence in theuranyl–α-hydroxycarboxylate systems that can be quantitatively described by the Stern–Volmer equation. As a result of the strong quenching it is not possible to detect fluorescence spectra for these complexes using TRLFS. The UO22+(aq) concentration calculated from the Stern–Volmer equation was used to determine equilibrium constants which are in good agreement with those obtained by potentiometry and NMR spectroscopy. No quenching was observed in the α-aminoisobutyrate system and their fluorescence spectra could be deconvoluted into components for the different species present. The following stability constants result from our TRLFS experiments: a) for the glycolate system log βUO₂(HOCH₂COO)⁺=2.52±0.20, b) for the α-hydroxyisobutyrate system log βUO₂[HOC(CH₃)₂COO]⁺=3.40±0.21, and c) for the α-aminoisobutyrate system logβUO₂[NH₃C(CH₃)₂COO]²⁺=1.30±0.10 and log βUO₂[NH₃C(CH₃)₂COO]₂²⁺=2.07±0.25. An increase of the fluorescence intensity connected with a red shift of the fluorescence emission spectra was observed in the system uranyl–α-aminoisobutyric acid. Fluorescence lifetimes and spectra were obtained for UO22+, UO2[NH3C(CH3)2COO]2+, and UO2[NH3C(CH3)2COO]22+. Uranium LIII-edge EXAFS measurements yielded an U-Oeq distance of 2.40 to 2.43 Å in the pH range from 2 to 4 in the α-hydroxyisobutyrate system showing a dominant bidentate coordination via the oxygens of the carboxylic group. Slightly shorter U-Oeq distances of 2.40 to 2.38 Å and no evidence for U-C distances around 2.90 Å in the glycolate system in this pH range may indicate a monodentate coordinated ligand via one oxygen from the carboxylic group. The decrease in the U-Oeq distance of the equatorial oxygens in both systems to 2.36-2.37 Å at pH values ≥5 is a strong indication for the formation of a chelate complex due to the deprotonation of the α-OH-group of the ligand. In the glycolate system in the pH range 5.5 to 11, the EXAFS spectrum showed evidence of U-U interaction at 3.81 Å indicating the formation of dimeric species.

The hydrolysis of dioxouranium(VI) investigated using EXAFS and 17O-NMR

The structure of dioxouranium(VI) as a function of pH at different (CH3)4N-OH concentrations has been investigated with the aid of U LIII-edge EXAFS. Polynuclear hydroxo species were identified by an U-U interaction at 3.808 Å at pH = 4.1. The precipitate formed at pH = 7 has a schoepite like structure. In solution at high pH [0.5 M (CH3)4N-OH], the EXAFS data are consistent with the formation of a monomeric four coordinated uranium(VI) hydroxide complex UO2(OH)42- of octahedral geometry. The first shell contains two O atoms with a U=O distance of 1.830 Å, and four O atoms were identified at a U-O distance of 2.265 Å. In strong alkaline solutions [>1 M (CH3)4N)-OH], 17O-NMR spectra indicate the presence of two species, presumably UO2(OH)42- and UO2(OH)53-, the latter in low concentration, which are in rapid equilibrium with one another at 268 K in aqueous solution.

Laser and X-ray spectroscopic studies of uranium-calcite interface phenomena

Abstract Calcite (CaCO 3 ) was treated with uranyl solutions of different concentrations in perchlorate medium at an initial pH of 6. X-ray photoelectron spectroscopy (XPS) measurements showed that there is no physi- or chemisorption of uranyl perchlorate molecules at the mineral surfaces and no diffusion of ClO 4 − anions into the mineral grains. We conclude this, because no chlorine atoms could be detected by XPS measurements. A significant amount of uranyl hydroxides is formed at the mineral surfaces. In addition, up to 20% of the calcium ions are exchanged by uranyl ions at the calcite surface. The inter-atomic uranium-oxygen distances of the reaction products at the mineral surface were determined using extended X-ray absorption fine structure (EXAFS) spectroscopy. In addition, the contacting solutions were studied by time-resolved laser-induced fluorescence spectroscopy (TRLFS). A new uranyl species was found in solution, which is also present in seepage waters from mine-tailing piles.

Solution coordination chemistry of uranium in the binary UO22+-SO42- and the ternary UO22+-SO42--OH- system

The structure and reaction dynamics in the systems UO22+-SO42- and UO22+-SO42--OH- were investigated using EXAFS and 17O-NMR spectroscopy. Uranium LIII edge EXAFS indicated a bidentate coordination mode of sulfate to uranyl. In solution, this is characterized by an U-S distance of 3.11 Å. Approximately 5 oxygen atoms were observed in the equatorial plane at 2.39-2.43 Å. The kinetics in the binary uranyl sulfate system can be described by four dominant exchange reactions: (1) UO22+ + SO42- ⇔ UO2SO4 (k1), (2) U*O22+ + UO2SO4 ⇔ U*O2SO4 + UO22+ (k2), (3) UO22+ + UO2(SO4)22- ⇔ 2 UO2SO4 (k3), and (4) UO2SO4 + SO42- ⇔ UO2(SO4)22- (k4). These reactions have rate constants indicating that the exchange is not of the simple Eigen-Wilkins type. Ternary uranyl sulfate hydroxide species were characterized by their 17O chemical shift and by potentiometry. There are no separate signals for the possible isomers of the ternary species indicating that they are in fast exchange with each other.