Insights into the interactions of cyanobacteria with uranium

Due to various activities associated with nuclear industry, uranium is migrated to aquatic environments like groundwater, ponds or oceans. Uranium forms stable carbonate complexes in the oxic waters of pH 7–10 which results in a high degree of uranium mobility. Microorganisms employ various mechanisms which significantly influence the mobility and the speciation of uranium in aquatic environments. Uranyl bioremediation studies, this far, have generally focussed on low pH conditions and related to adsorption of positively charged UO₂ ²⁺ onto negatively charged microbial surfaces. Sequestration of anionic uranium species, i.e. [UO₂(CO₃) ₂ ^2? ] and [UO₂(CO₃) ₃ ^4? ] onto microbial surfaces has received only scant attention. Marine cyanobacteria are effective metal adsorbents and represent an important sink for metals in aquatic environment. This article addresses the cyanobacterial interactions with toxic metals in general while stressing on uranium. It focusses on the possible mechanisms employed by cyanobacteria to sequester uranium from aqueous solutions above circumneutral pH where negatively charged uranyl carbonate complexes dominate aqueous uranium speciation. The mechanisms demonstrated by cyanobacteria are important components of biogeochemical cycle of uranium and are useful for the development of appropriate strategies, either to recover or remediate uranium from the aquatic environments.     

Interaction between uranium and the cytochrome c 3 of Desulfovibrio desulfuricans strain G20

Cytochrome c₃ of Desulfovibrio desulfuricans strain G20 is an electron carrier for uranium (VI) reduction. When D. desulfuricans G20 was grown in medium containing a non-lethal concentration of uranyl acetate (1 mM), the rate at which the cells reduced U(VI) was decreased compared to cells grown in the absence of uranium. Western analysis did not detect cytochrome c₃ in periplasmic extracts from cells grown in the presence of uranium. The expression of this predominant tetraheme cytochrome was not detectably altered by uranium during growth of the cells as monitored through a translational fusion of the gene encoding cytochrome c₃ (cycA) to lacZ. Instead, cytochrome c₃ protein was found tightly associated with insoluble U(IV), uraninite, after the periplasmic contents of cells were harvested by a pH shift. The association of cytochrome c₃ with U(IV) was interpreted to be non-specific, since pure cytochrome c₃ adsorbed to other insoluble metal oxides, including cupric oxide (CuO), ferric oxide (Fe₂O₃), and commercially available U(IV) oxide.An erratum to this article can be found at     

Respiratory Levels and Adaptations in Four Freshwater Species of Gammarus (Crustacea: Amphipoda)

Respiration of four freshwater species of the amphipod crustacean Gammarus: G. fossarum, G. lacustris (river and lake), G. pulex and G. roeseli were measured in a closed, stirred respirometric chamber with a micro-electrode. Oxygen consumption, expressed as weight-specific oxygen uptake (R_s) in relation to decreasing oxygen concentration, varied at air saturation from 0.86 (G. lacustris, lake) to 2.06 μl O₂ mg⁻¹ AFDW h⁻¹ (G. pulex). R_s also differed intra-specifically among the two populations of G. lacustris. G. lacustris (river), G. pulex and G. roeseli expressed moderate ability to regulate their oxygen consumption at decreasing oxygen concentrations, whereas the regulation ability was higher in G. lacustris (lake) and in G. fossarum, which maintain high oxygen uptake at oxygen levels >2 mg O₂ l⁻¹. All four Gammarus species are partial regulators in response to variations in oxygen concentration. The differences between species are considered too small to account for their natural distributions. It appears that the tolerances of Gammarus species to organic pollution depend only in part on oxygen conditions.     

GPCR-induced dissociation of G-protein subunits in early stage signal transduction

G-protein coupled receptors (GPCRs) form a ternary complex of agonist, receptor and G-proteins during primary signal transduction at the cell membrane. Downstream signalling is thought to be preceded by the process of dissociation of Gα and Gβγ subunits, thus exposing new surfaces to interact with downstream effectors. We demonstrate here for the first time, the dissociation of heterotrimeric G-protein subunits (i.e., Gα and Gβγ) following agonist-induced GPCR (α_2A-adrenergic receptor; α_2A-AR) activation in a cell-free assay system. α_2A-AR membranes were reconstituted with the G-proteins (±hexahistidine-tagged) Gα_i1 and Gβ₁γ₂ and functional signalling was determined following activation of the reconstituted receptor:G-protein complex with the potent agonist UK-14304, and [³⁵S]GTPγS. In the presence of Ni²⁺-coated agarose beads, the activated his-tagged Gα_i1his-[³⁵S]GTPγS complex was captured on the Ni²⁺-presenting surface. When his-tagged Gβ₁γ₂ (Gβ₁γ₂his) was used with Gα_i1, the [³⁵S]GTPγS-bound Gα_i1 was not present on the Ni²⁺-coated beads, but rather, it was separated from the β₁γ₂(his)-beads, demonstrating receptor-induced dissociation of Gα and Gβγ subunits. Treatment of the reconstituted α_2A-AR membranes containing Gβ₁γ₂his:Gα_i1 with imidazole confirmed the specificity for the Ni²⁺:G-protein surface dissociation of Gα_i1 from Gβ₁γ₂his. These data demonstrate for the first time, the complete dissociation of the G-protein subunits and extend observations on the role of G-proteins in the assembly and disassembly of the ternary complex in the primary events of GPCR signalling.     

A p and N NMR study of the extraction of uranyl nitrate by Di-2-ethylhexyl phosphoric acid

Low temperature ³¹P and ¹⁵N NMR spectroscopy was used to investigate the species forming in the organic layer following the extraction of uranium from nitric acid solutions with di-2-ethylhexyl phosphoric acid. It was found that uranium is extracted from neutral solutions as the 1:2 complex UO₂A₂ regardless of what anion is present. For dilute nitric acid solutions, the uranium is extracted both as associated and mixed nitrato species. As the nitric acid concentration of the aqueous layer increases, the mixed nitrato complex, UO2(NO₃)A·HA, becomes predominant.     

Bioleaching of uranium from low grade black schists by <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis>

Summary The feasibility of bacterial recovery of uranium from the low grade black schists occurring in the Okcheon district, South Korea, was investigated. Following the introduction of Acidithiobacillus ferrooxidans, 80% of the uranium could be extracted from the schists, which contain 0.01% U₃O₈ by weight, within 60 h at a pulp density of 100 g-ore/l. Only 18% of the uranium was extracted without microbial activity. The uranium-leaching efficiency was not greatly affected by the addition of Fe²⁺ in the range of 5–9 g/l, and the leaching efficiency of uranium from the schists by A. ferrooxidanscould be efficiently maintained at high pulp densities (up to 500 g-ore/l).     

Effects of various metabolites on two phosphoglucomutase allozyme activities from Drosophila melanogaster

The effects of various metabolites on the two most common phosphoglucomutase allozymes (PGM^A and PGM^B) in Drosophila melanogaster have been investigated in vitro. 2,3-Diphosphoglycerate (2,3DPG) inhibited PGM^A and PGM^B to the same degree in the presence of 25 µM glucose-1,6-diphosphate (G1, 6P₂). However a higher concentration of G1,6P₂ partially reversed the inhibition of PGM^A exerted by 2,3DPG, so that in the presence of 150 µM G1,6P₂ the inhibition of PGM^A was half that of PGM^B at pH 6.0. Glycerol-3-phosphate (G3P) had no significant effect at pH 7.4 but exerted an activating effect at pH 6.0 which was more pronounced in the case of PGM^B. ATP, citrate, and fructose-1, 6-diphosphate (F1,6P₂) inhibited both PGM^A and PGM^B. The differences found in vitro between these two allozymes can have a significant impact on in vivo function and, therefore, on the maintenance of PGM polymorphism in experimental populations of D. melanogaster studied in the laboratory.     

Effects of Nucleotides on [3H]Bradykinin Binding in Guinea Pig: Further Evidence for Multiple B2 Receptor Subtypes

Abstract: We have suggested recently the existence of three subtypes of B₂ bradykinin receptors in tissues of guinea pigs. We have classified these B₂ bradykinin receptors into B_2a, B_2b, and B_2c subtypes depending on their affinity for various bradykinin antagonists. Because the actions of bradykinin in different cell systems appear to be both dependent on and independent of G proteins, we sought to determine whether the binding of [³H]bradykinin to the B₂ subtypes is sensitive to guanine nucleotides and, therefore, possibly coupled to G proteins. In the ileum, where we have demonstrated B_2a and B_2b subtypes, specific [³H]bradykinin binding was reduced with GDP (100 μM) and the nonmetabolized analogue of GTP, guanyl-5′-yl-imidodiphosphate (GppNHp; 100 μM). Competition studies with bradykinin and with [Hyp³]-bradykinin, which shows approximately 20-fold greater selectivity for the B_2a subtype than bradykinin, were performed in the presence or absence of GppNHp (100 μM). The competition experiments demonstrated that binding to the B_2a subtype, which has higher affinity for [Hyp³]-bradykinin and bradykinin than the B_2b subtype, was lost in the presence of GppNHp, whereas binding to the B_2b subtype was unaffected. In contrast, GppNHp (100 μM) and GDP (100 μM) failed to alter specific [³H]bradykinin binding to B_2b and B_2c subtypes in lung. [³H]Bradykinin binding was unaffected by AMP, ADP, ATP, and GMP (100 μM each). Based on this evidence, we suggest that the B_2a bradykinin subtype is coupled to G proteins. The B_2b and B_2c subtypes are either not coupled to G proteins, or may be coupled to the G^o-type GTP binding proteins, which have been suggested to be less sensitive to guanine nucleotides. These data provide further evidence for three subtypes of B₂-type bradykinin receptors in guinea pig.     

Uranium phosphate biomineralization by fungi

Geoactive soil fungi were investigated for phosphatase‐mediated uranium precipitation during growth on an organic phosphorus source. Aspergillus niger and Paecilomyces javanicus were grown on modified Czapek–Dox medium amended with glycerol 2‐phosphate (G2P) as sole P source and uranium nitrate. Both organisms showed reduced growth on uranium‐containing media but were able to extensively precipitate uranium and phosphorus‐containing minerals on hyphal surfaces, and these were identified by X‐ray powder diffraction as uranyl phosphate species, including potassium uranyl phosphate hydrate (KPUO₆.3H₂O), meta‐ankoleite [(K_1.7Ba_0.2)(UO₂)₂(PO₄)₂.6H₂O], uranyl phosphate hydrate [(UO₂)₃(PO₄)₂.4H₂O], meta‐ankoleite (K(UO₂)(PO₄).3H₂O), uramphite (NH₄UO₂PO₄.3H₂O) and chernikovite [(H₃O)₂(UO₂)₂(PO₄)₂.6H₂O]. Some minerals with a morphology similar to bacterial hydrogen uranyl phosphate were detected on A. niger biomass. Geochemical modelling confirmed the complexity of uranium speciation, and the presence of meta‐ankoleite, uramphite and uranyl phosphate hydrate between pH 3 and 8 closely matched the experimental data, with potassium as the dominant cation. We have therefore demonstrated that fungi can precipitate U‐containing phosphate biominerals when grown with an organic source of P, with the hyphal matrix serving to localize the resultant uranium minerals. The findings throw further light on potential fungal roles in U and P biogeochemistry as well as the application of these mechanisms for element recovery or bioremediation.     

Effects of water hardness and alkalinity on the toxicity of uranium to a tropical freshwater hydra (Hydra viridissima)

In tropical Australian freshwaters, uranium (U) is of potential ecotoxicological concern, largely as a consequence of mining activities. Although the toxicity of uranium to Australian freshwater biota is comprehensive, by world standards, few data are available on the effects of physicochemical variables, such as hardness, alkalinity, pH and organic matter, on uranium speciation and bioavailability. This study determined the individual effects of water hardness (6.6, 165 and 330 mg l^-1 as CaCO₃) and alkalinity (4.0 and 102 mg l^-1 as CaCO₃), at a constant pH (6.0), on the toxicity (96 h population growth) of uranium to Hydra viridissima (green hydra). A 50-fold increase in hardness (Ca and Mg concentration) resulted in a 92% (two-fold) decrease in the toxicity of uranium to H. viridissima [i.e. an increase in the EC₅₀ value and 95% confidence interval from 114 (107-121) to 219 (192-246) µg l^-1]. Conversely, at a constant hardness (165 mg l-1 as CaCO₃), the toxicity of uranium to H. viridissima was not significantly (P &gt; 0.05) affected by a 25-fold increase in alkalinity (carbonate concentration) [i.e. EC₅₀ values of 177 (166-188) and 171 (150-192) µg l^-1 at 4.0 and 102 mg l^-1 as CaCO₃, respectively]. A knowledge of the relationship between water chemistry variables, including hardness and alkalinity, and uranium toxicity is useful for predicting the potential ecological detriment in aquatic systems, and can be used to relax national water quality guidelines on a site-specific basis.     

Identification of platelet membrane thrombospondin binding molecules using an anti-thrombospondin antibody

A rat monoclonal IgG_2a antibody, 5G11, was raised against native human platelet thrombospondin (TSP). Western blot analysis revealed that 5G11 bound (i) to TSP before and after disulfide reduction, and (ii) to a 15-kDa fragment released after prolonged trypsin digestion. Crossed immunoelectrophoresis confirmed that the binding epitope was expressed in the presence of Ca²⁺ and after treatment of TSP with EDTA. Since 5G11 had no effect on platelet aggregation, the antibody was used to immunoprecipitate Ca²⁺-dependent and Ca²⁺-independent TSP-binding molecules on the surface of thrombin-activated surface-labeled ¹²⁵I-platelets. The experimental basis was that ligand-receptor interactions are of high affinity and that anti-ligand antibodies should precipitate the ligand-receptor complex. With platelets activated in the presence of EDTA, 5G11 predominantly precipitated a ¹²⁵I-labeled band of M_r 88 000, identified as glycoprotein (GP) IV. In contrast, in the presence of 2 mM Ca²⁺ and 1 mM Mg²⁺, 5G11 precipitated a complex of five radiolabeled proteins, among which GPIIb, GPIIIa and GPIV were the most prominent.     

Relationship between growth parameters of the amphipod Gammarus zaddachi (Sexton 1912) and the permeable body surface area determined by the acid-base titration method

More than 10000 specimens of the amphipod Gammarus zaddachiwere collected from the River Hunte (Huntebrück, Germany) in January, April, July and September 1999 and Jan 2000 to analyse relationships between body-length distributions, growth and the permeable body surface area (from which ions can enter the cell) as indicated by the acid–base titration method. Thus, body wet weights (BWWs), dry weights (BDWs), length (BL), surface area and specific surface area were measured. Mean BWWs were 56.0 mg, BDWs 11.1 mg and BLs 16.4 mm. The relationship between BDW and BL could be successfully described by the power-function: BDW=0.0029×BL ^2.88, probably indicating that the body volume of the test animals can be approximately expressed by an `equal-effectiveness ball'. The body-length distributions were analysed using two combined normal-distribution equations, suggesting that the population of G. zaddachi at Huntebrück was composed of two generations. A sigmoid logistic equation was applied to estimate the body length-based growth. It can be inferred that the reproduction must have occurred around October to December 1998 and that the size of juveniles in the brood pouch was 0.1 mm. The permeable body surface area (ABSA) of G. zaddachi was measured by applying the acid–base titration method, with an average measurement accuracy of 13.7%. ABSA ranged from 17.1 to 236.9 cm² animal^–1 in a variety of the body lengths studied. The specific surface areas (SSA), equivalent to the ratio of permeable body surface area to body volume, decreased from 14.2 to 8.5 cm² mg^–1 (dry wt) as BL increased from 8.1 to 18.6 mm and then remained nearly constant up to 24.1 mm, indicating a single exponential decay relationship between SSA and BL. Mean body length (BL _mean) of G. zaddachi collected simultaneously at Huntebrück increased seasonally from 0.1 mm in autumn–winter to 17.6 in next winter–spring, resulting in an increase of ABSA but a decrease of SSA. Our present results provide a sound and necessary basis for a quantitative study not only on the age-dependent biological/physiological conditions but also on size-dependent bioconcentration of metal by gammaridean amphipods.     

The Arabidopsis heterotrimeric G‐protein β subunit,AGB1, is required for guard cell calcium sensing and calcium‐induced calcium release

Cytosolic calcium concentration ([Ca²⁺]_cyt) and heterotrimeric G‐proteins are universal eukaryotic signaling elements. In plant guard cells, extracellular calcium (Ca_o) is as strong a stimulus for stomatal closure as the phytohormone abscisic acid (ABA), but underlying mechanisms remain elusive. Here, we report that the sole Arabidopsis heterotrimeric Gβ subunit, AGB1, is required for four guard cell Ca_o responses: induction of stomatal closure; inhibition of stomatal opening; [Ca²⁺]_cyt oscillation; and inositol 1,4,5‐trisphosphate (InsP3) production. Stomata in wild‐type Arabidopsis (Col) and in mutants of the canonical Gα subunit, GPA1, showed inhibition of stomatal opening and promotion of stomatal closure by Ca_o. By contrast, stomatal movements of agb1 mutants and agb1/gpa1 double‐mutants, as well as those of the agg1agg2 Gγ double‐mutant, were insensitive to Ca_o. These behaviors contrast with ABA‐regulated stomatal movements, which involve GPA1 and AGB1/AGG3 dimers, illustrating differential partitioning of G‐protein subunits among stimuli with similar ultimate impacts, which may facilitate stimulus‐specific encoding. AGB1 knockouts retained reactive oxygen species and NO production, but lost YC3.6‐detected [Ca²⁺]_cyt oscillations in response to Ca_o, initiating only a single [Ca²⁺]_cyt spike. Experimentally imposed [Ca²⁺]_cyt oscillations restored stomatal closure in agb1. Yeast two‐hybrid and bimolecular complementation fluorescence experiments revealed that AGB1 interacts with phospholipase Cs (PLCs), and Ca_o induced InsP3 production in Col but not in agb1. In sum, G‐protein signaling via AGB1/AGG1/AGG2 is essential for Ca_o‐regulation of stomatal apertures, and stomatal movements in response to Ca_o apparently require Ca²⁺‐induced Ca²⁺ release that is likely dependent on Gβγ interaction with PLCs leading to InsP3 production.     

Glucose-6-phosphate dehydrogenase-dependent hydrogen peroxide production is involved in the regulation of plasma membrane H+-ATPase and Na+/H+ antiporter protein in salt-stressed callus from Carex moorcroftii

Glucose‐6‐phosphate dehydrogenase (G6PDH) is important for the activation of plant resistance to environmental stresses, and ion homeostasis is the physiological foundation for living cells. In this study, we investigated G6PDH roles in modulating ion homeostasis under salt stress in Carex moorcroftii callus. G6PDH activity increased to its maximum in 100 mM NaCl treatment and decreased with further increased NaCl concentrations. K⁺/Na⁺ ratio in 100 mM NaCl treatment did not exhibit significant difference compared with the control; however, in 300 mM NaCl treatment, it decreased. Low‐concentration NaCl (100 mM) stimulated plasma membrane (PM) H⁺‐ATPase and NADPH oxidase activities as well as Na⁺/H⁺ antiporter protein expression, whereas high‐concentration NaCl (300 mM) decreased their activity and expression. When G6PDH activity and expression were reduced by glycerol treatments, PM H⁺‐ATPase and NADPH oxidase activities, Na⁺/H⁺ antiporter protein level and K⁺/Na⁺ ratio dramatically decreased. Simultaneously, NaCl‐induced hydrogen peroxide (H₂O₂) accumulation was abolished. Exogenous application of H₂O₂ increased G6PDH, PM H⁺‐ATPase and NADPH oxidase activities, Na⁺/H⁺ antiporter protein expression and K⁺/Na⁺ ratio in the control and glycerol treatments. Diphenylene iodonium (DPI), the NADPH oxidase inhibitor, which counteracted NaCl‐induced H₂O₂ accumulation, decreased G6PDH, PM H⁺‐ATPase and NADPH oxidase activities, Na⁺/H⁺ antiporter protein level and K⁺/Na⁺ ratio. Western blot result showed that G6PDH expression was stimulated by NaCl and H₂O₂, and blocked by DPI. Taken together, G6PDH is involved in H₂O₂ accumulation under salt stress. H₂O₂, as a signal, upregulated PM H⁺‐ATPase activity and Na⁺/H⁺ antiporter protein level, which subsequently resulted in the enhanced K⁺/Na⁺ ratio. G6PDH played a central role in the process.     

Conformational preferences of modified nucleoside N(2)-methylguanosine (m(2)G) and its derivative N(2), N(2)-dimethylguanosine (m(2)(2)G) occur at 26th position (hinge region) in tRNA

Conformational preferences of the modified nucleosides N²-methylguanosine (m²G) and N², N²-dimethylguanosine (m₂²G) have been studied theoretically by using quantum chemical perturbative configuration interaction with localized orbitals (PCILO) method. Automated complete geometry optimization using semiempirical quantum chemical RM1, along with ab initio molecular orbital Hartree–Fock (HF-SCF), and density functional theory (DFT) calculations has also been made to compare the salient features. Single-point energy calculation studies have been made on various models of m²G26:C/A/U44 and m₂²G26:C/A/U44. The glycosyl torsion angle prefers “syn” (χ = 286°) conformation for m²G and m₂²G molecules. These conformations are stabilized by N(3)–HC2′ and N(3)–HC3′ by replacing weak interaction between O5′–HC(8). The N²-methyl substituent of (m²G26) prefers “proximal” or s-trans conformation. It may also prefer “distal” or s-cis conformation that allows base pairing with A/U44 instead of C at the hinge region. Thus, N²-methyl group of m²G may have energetically two stable s-trans m²G:C/A/U or s-cis m²G:A/U rotamers. This could be because of free rotations around C–N bond. Similarly, N², N²-dimethyl substituent of (m₂²G) prefers “distal” conformation that may allow base pairing with A/U instead of C at 44th position. Such orientations of m²G and m₂²G could play an important role in base-stacking interactions at the hinge region of tRNA during protein biosynthesis process.     

Uranium luminescence in La2Zr2O7: effect of concentration and annealing temperature

The speciation of a particular element in any given matrix is a prerequisite to understanding its solubility and leaching properties. In this context, speciation of uranium in lanthanum zirconate pyrochlore (La₂Zr₂O₇ = LZO), prepared by a low‐temperature combustion route, was carried out using a simple photoluminescence lifetime technique. The LZO matrix is considered to be a potential ceramic host for fixing nuclear and actinide waste products generated during the nuclear fuel cycle. Special emphasis has been given to understanding the dynamics of the uranium species in the host as a function of annealing temperature and concentration. It was found that, in the LZO host, uranium is stabilized as the commonly encountered uranyl species (UO₂²⁺) up to a heat treatment of 500 °C at the surface. Above 500 °C, the uranyl ion is diffused into the matrix as the more symmetric octahedral uranate species (UO₆^6–). The uranate ions thus formed replace the six‐coordinated ‘Zr’ atoms at regular lattice positions. Further, it was observed that concentration quenching takes place beyond 5 mol% of uranium doping. The mechanism of the quenching was found to be a multipolar interaction. Copyright © 2016 John Wiley & Sons, Ltd.     

The ATPase activity of the <Emphasis Type="Italic">G2alt</Emphasis> gene encoding an aluminium tolerance protein from <Emphasis Type="Italic">Anoxybacillus gonensis</Emphasis> G2

The G2ALT gene was cloned and sequenced from the thermophilic bacterium Anoxybacillus gonensis G2. The gene is 666 bp long and encodes a protein 221 amino acids in length. The gene was overexpressed in E. coli and purified to homogeneity and biochemically characterized. The enzyme has a molecular mass of 24.5 kDa and it could be classified as a member of the family of bacterial aluminium resistance proteins based on homology searches. When this fragment was expressed in E. coli, it endowed E. coli with Al tolerance to 500 μM. The purified G2ALT protein is active at a broad pH range (pH 4.0–10.0) and temperature range (25°C–80°C) with optima of 6.0 and the apparent optimal temperature of 73°C respectively. Under optimal conditions, G2ALT exhibited a low ATPase activity with K _m ⁻ and V _max ⁻ values of 10±0.55 μM and 26.81±0.13 mg Pi released/min/mg enzyme, respectively. The ATPase activity of G2ALT requires Mg²⁺ and Na⁺ ions, while Zn²⁺ and Al³⁺ stimulate the activity. Cd²⁺ and Ag⁺ reduced the activity and Li⁺, Cu²⁺, and Co²⁺ inhibited the activity. Known inhibitors of most ATPases, like such as β-mercaptoethanol and ouabain, also inhibited the activity of the G2ALT. These biochemical characterizations suggested that G2ALT belongs to the PP-loop ATPase superfamily and it can be responsible for aluminium tolerance in A. gonensis G2.     

Studies on the growth ofThiobacillus ferrooxidans

Summary Uranyl sulphate (0.2–0.9 mM) inhibited ferrous iron oxidation by growing cultures ofThiobacillus ferrooxidans. The addition of 5–100 mM uranium to the cultures caused immediate cessation of carbon dioxide fixation, rapid loss of viability and gradual depression of ferrous iron oxidation. Virtually no uranium was found in washed cells grown in the presence of subtoxic to toxic amounts of uranyl sulphate. Uranium-poisoned organisms appeared plasmolyzed in electron micrographs. Cultures tolerant to 5 mM UO₂ ²⁺ were develoepd by successive subculturing in increased uranium concentrations. The tolerance was maintained during subculturing in uranium-free medium. Frequency of mutants resistant to 1.0 and 1.5 mM UO₂ ²⁺ was 1 per 1.3×10⁶ and 1 per 9.0×10⁸, respectively. The frequency was increased in the presence of 15–150 mM nickel, zinc and manganese. In liquid cultures, bivalent cations and EDTA alleviated the toxicity of 2 mM uranyl sulphate.     

Recovery of uranium by immobilized microorganisms

Summary Some attempts were made to recover uranium from sea and fresh water using immobilized Streptomyces viridochromogenes and Chlorella regularis cells. The cells immobilized in polyacrylamide gel have the most favorable features for uranium recovery; high adsorption ability, good mechanical properties, and applicability in a column system. The adsorption of uranium by the immobilized cells is not affected by the pH values between 4 and 9. These results show that uranium adsorption becomes independent of pH after immobilization. The amounts of uranium adsorbed by the immobilized cells increased linearly with temperature, suggesting that the adsorption of uranium by the immobilized cells is an endothermic reaction. The immobilized cells can recover uranium almost quantitatively from both fresh and sea water containing uranium, and almost all uranium adsorbed is desorbed with a solution of Na₂CO₃. Thus the immobilized cells of Streptomyces and Chlorella can be used repeatedly in adsorption-desorption process.Studies on the Accumulation of Heavy Metal Elements in Biological Systems. XXI     

A single amino acid substitution in the MurF UDP‐MurNAc‐pentapeptide synthetase renders Streptococcus pneumoniae dependent on CO2 and temperature

The respiratory tract pathogen Streptococcus pneumoniae encounters different levels of environmental CO₂ during transmission, host colonization and disease. About 8% of all pneumococcal isolates are capnophiles that require CO₂‐enriched growth conditions. The underlying molecular mechanism for caphnophilic behaviour, as well as its biological function is unknown. Here, we found that capnophilic S. pneumoniae isolates from clonal complex (CC) 156 (i.e. Spain^9V‐3 ancestry) and CC344 (i.e. Norway^NT‐42 ancestry) have a valine at position 179 in the MurF UDP‐MurNAc‐pentapeptide synthetase. At ≤ 30°C, the growth characteristics of capnophilic and non‐capnophilic CC156 strains were equal, but at > 30°C growth and survival of MurF^V179 strains was dependent on > 0.1% CO₂‐enriched conditions. Expression of MurF^V179 in S. pneumoniae R6 and G54 rendered these, otherwise non‐capnophilic strains, capnophilic. Time‐lapse microscopy revealed that a capnophilic CC156 strain undergoes rapid autolysis upon exposure to CO₂‐poor conditions at 37°C, and staining with fluorescently labelled vancomycin showed a defect in de novo cell wall synthesis. In summary, in capnophilic S. pneumoniae strains from CC156 and CC344 cell wall synthesis is placed under control of environmental CO₂ levels and temperature. This mechanism might represent a novel strategy of the pneumococcus to rapidly adapt and colonize its host under changing environmental conditions.