Microbial Cells as Biosorbents for Heavy Metals: Accumulation of Uranium by Saccharomyces cerevisiae and Pseudomonas aeruginosa

Uranium accumulated extracellularly on the surfaces of Saccharomyces cerevisiae cells. The rate and extent of accumulation were subject to environmental parameters, such as pH, temperature, and interference by certain anions and cations. Uranium accumulation by Pseudomonas aeruginosa occurred intracellularly and was extremely rapid (<10 s), and no response to environmental parameters could be detected. Metabolism was not required for metal uptake by either organism. Cell-bound uranium reached a concentration of 10 to 15% of the dry cell weight, but only 32% of the S. cerevisiae cells and 44% of the P. aeruginosa cells within a given population possessed visible uranium deposits when examined by electron microscopy. Rates of uranium uptake by S. cerevisiae were increased by chemical pretreatment of the cells. Uranium could be removed chemically from S. cerevisiae cells, and the cells could then be reused as a biosorbent.     

Rapid biodegradation and decolorization of Direct Orange 39 (Orange TGLL) by an isolated bacterium <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> strain BCH

A newly isolated novel bacterium from sediments contaminated with dyestuff was identified as Pseudomonas aeruginosa strain BCH by 16S rRNA gene sequence analysis. The bacterium was extraordinarily active and operative over a wide rage of temperature (10–60°C) and salinity (5–6%), for decolorization of Direct Orange 39 (Orange TGLL) at optimum pH 7. This strain was capable of decolorizing Direct Orange 39; 50 mg l⁻¹ within 45 ± 5 min, with 93.06% decolorization, while maximally it could decolorize 1.5 g l⁻¹ of dye within 48 h with 60% decolorization. Analytical studies as, UV–Vis spectroscopy, FTIR, HPLC were employed to confirm the biodegradation of dye and formation of new metabolites. Induction in the activities of lignin peroxidases, DCIP reductase as well as tyrosinase was observed, indicating the significant role of these enzymes in biodegradation of Direct Orange 39. Toxicity studies with Phaseolus mungo and Triticum aestivum revealed the non-toxic nature of degraded metabolites.     

Effects of selected physical and chemical parameters on uranium uptake by the bacterium Chryseomonas MGF-48

The effect of selected physical and chemical parameters, including media composition, uranium concentration, and metabolic inhibitors, on uranium uptake from aqueous solution by Chryseomonas sp. MGF-48, a bacterium isolated from electroplating effluent, was analysed. It was found that 198 mg/g (dry weight) of uranium was taken up from a 150 ppm (mg/l) solution of uranium when cells were maintained under starvation conditions. Uranium was released from the cells by addition of sodium carbonate. Uptake of uranium was reduced when cells were subjected to pretreatment with ultraviolet (u.v.) light (263.7 nm, 10 min) or heat (100 °C, 15 min), i.e., 99.8 and 57.5 mg/g, respectively. Addition of 2, 4-dinitrophenol (10 mM) or sodium azide (0.1%) resulted in decreased uptake (129 and 123 mg/g, respectively), whereas addition of glycerol-2-phosphate (G-2-P) resulted in deposition of uranium. Maximum uptake of uranium occurred at pH 6.5. Uranium uptake in the presence of selected carbohydrates decreased as follows: xylose > arabinose > mannose > maltose > glucose. When Chryseomonas sp. MGF-48 was exposed to 5, 50, or 100 mg/l uranium prior to uptake experiments, a decrease in uranium uptake was noted. Furthermore, immobilization of the bacterial cells in a calcium-alginate medium reduced the efficiency of uptake, yielding only 60% uranium uptake. It is concluded that uranium uptake and accumulation by this bacterium involves both metabolism-independent absorption and metabolism-dependent absorption, the former being considered to be more efficient.     

Characterization of a metal resistant Pseudomonas sp. isolated from uranium mine for its potential in heavy metal (Ni2+, Co2+, Cu2+, and Cd2+) sequestration

Heavy metal sequestration by a multimetal resistant Pseudomonas strain isolated from a uranium mine was characterized for its potential application in metal bioremediation. 16S rRNA gene analysis revealed phylogenetic relatedness of this isolate to Pseudomonas fluorescens. Metal uptake by this bacterium was monophasic, fast saturating, concentration and pH dependent with maximum loading of 1048 nmol Ni²⁺ followed by 845 nmol Co²⁺, 828 nmol Cu²⁺ and 700 nmol Cd²⁺ mg^?1 dry wt. Preferential metal deposition in cell envelope was confirmed by TEM and cell fractionation. FTIR spectroscopy and EDX analysis revealed a major role of carboxyl and phosphoryl groups along with a possible ion exchange mechanism in cation binding. Binary system demonstrated selective metal binding affinity in the order of Cu²⁺ > Ni²⁺ > Co²⁺ > Cd²⁺. A comparison with similar metal uptake reports considering live bacteria strongly indicated the superiority of this strain in metal sequestration, which could be useful for developing efficient metal removal system.     

Cloning,expression, purification,and characterization of recombinant flagellin isolated from <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

Pseudomonas aeruginosa as an opportunistic pathogen causes lethal infections in immunocompromised individuals. This bacterium possesses a polar flagellum made up of flagellin subunits. Flagella have important roles in motility, chemotaxis, and establishment of P. aeruginosa in acute phase of infections. Isolation, cloning, and expression of flagellin were aimed at in this study. Flagellin gene (fliC) of P. aeruginosa strain 8821M was isolated by PCR and cloned into a pET expression vector. The recombinant flagellin (46 kDa) was overexpressed as inclusion bodies (IBs). IBs were solubilized in guanidine hydrochloride (GuHCl) followed by affinity-purification and renatured using Ni²⁺-Sepharose resin. Recombinant flagellins reacted with the serum from a rabbit previously immunized with native flagellin. In addition, polyclonal antiserum raised against the recombinant flagellin was shown to significantly inhibit the cell motility of P. aeruginosa strain 8821M in vitro.     

Studies on Uranium Removal by the Extracellular Polysaccharide of a Pseudomonas aeruginosa Strain

Extracellular polysaccharide (EPS) produced by a Pseudomonas aeruginosa strain BU2 was characterized for its ability to remove uranium from aqueous solution. The EPS was acidic in nature and found as a potent biosorbent for uranium (U), showing pH dependence and fast saturating metal sorption, being maximum (985 mg U g^{? 1} EPS) at pH 5.0. The polymer showed enhanced uranium sorption capacity and affinity with increasing solution pH, suggesting a preferential sorption of monovalent uranyl hydroxide ions over the nonhydroxylated divalent species. Pseudo-first-order and pseudo-second-order kinetic models were applied to the experimental data, assuming that the external mass transfer limitations in the system can be neglected and biosorption is sorption controlled. Equilibrium metal binding showing conformity to the Freundlich model suggested a multilayer sorption involving specific binding sites with affinity distribution. The presence of two types of metal binding sites corresponding to strong and weak binding affinity was interpreted from the Scatchard model equation. Uranium sorption by EPS was unaffected or only slightly affected in the presence of several interfering cations and anions, except iron and thorium. Fourier transform infrared (FTIR) spectroscopy ascertained the strong binding of uranium with the carboxylic groups of uronic acids of bacterial EPS at pH 5.0, whereas at lower pH, amino and hydroxyl groups played a major role in metal binding.     

The cell wall amidase AmiB is essential for Pseudomonas aeruginosa cell division,drug resistance and viability

The physiological function of cell wall amidases has been investigated in several proteobacterial species. In all cases, they have been implicated in the cleavage of cell wall material synthesized by the cytokinetic ring. Although typically non‐essential, this activity is critical for daughter cell separation and outer membrane invagination during division. In Escherichia coli, proteins with LytM domains also participate in cell separation by stimulating amidase activity. Here, we investigated the function of amidases and LytM proteins in the opportunistic pathogen Pseudomonas aeruginosa. In agreement with studies in other organisms, ^PaAmiB and three LytM proteins were found to play crucial roles in P. aeruginosa cell separation, envelope integrity and antibiotic resistance. Importantly, the phenotype of amidase‐defective P. aeruginosa cells also differed in informative ways from the E. coli paradigm; ^PaAmiB was found to be essential for viability and the successful completion of cell constriction. Our results thus reveal a key role for amidase activity in cytokinetic ring contraction. Furthermore, we show that the essential function of ^PaAmiB can be bypassed in mutants activated for a Cpx‐like envelope stress response, suggesting that this signaling system may elicit the repair of division machinery defects in addition to general envelope damage.     

The Algicidal Characteristics of One Algae-Lysing FDT5 Bacterium on Microcystis aeruginosa

One strain of algicidal bacterium which can inhibit Harmful algal blooms (HABs), FDT5, was isolated from activated sludge and found to have good algicidal effects on Microcystis aeruginosa. It was revealed that: The FDT5 was a Gram-negative bacterium and identified as Ochrobactrum sp.; The greater the initial bacterial cell density, the faster the degradation of chlorophyll a.; The algicidal efficiency was evaluated at the most favorable conditions which were a temperature of 30–35°C, a pH of 7.6 and complete darkness; The FDT5 strain lysed Microcystis aeruginosa not directly but by secreting metabolites which could withstand high temperatures and pressure.     

Concentration dependent growth/non-growth linked kinetics of endosulfan biodegradation by <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

The rates of biodegradation of endosulfan by P. aeruginosa were determined with different initial endosulfan concentrations (10, 50, 100, 150, 200 and 250 mg l⁻¹) and different growth linked kinetic models were fitted at these concentrations. At 10 mg endosulfan l⁻¹, Monod no growth model was well fitted. Monod with growth model described the biodegradation pattern at an initial concentration of 50, 100 and 150 mg endosulfan l⁻¹. Significant increases of P. aeruginosa MN2B14 density in broth culture during incubation further support this result. Conversely, zero order kinetic model was well fitted into the biodegradation data if initial endosulfan concentration was ≥200 mg endosulfan l⁻¹. The kinetics of endosulfan biodegradation by P. aeruginosa MN2B14 in liquid broth was highly dependent upon its initial concentration. The results of this study could be employed for predicting the persistence of endosulfan in water environment containing P. aeruginosa as an endosulfan degrading bacterium.     

The effect of <Emphasis Type="Italic">Poterioochromonas</Emphasis> abundance on production of intra- and extracellular microcystin-LR concentration

Due to its capability for producing various microcystins, Microcystis aeruginosa is recognized as one of the most toxic, bloom-forming cyanobacteria. In this study, the fates of intra- and extracellular microcystin-LR (MC-LR) were investigated when the mixotrophic golden alga Poterioochromonas sp. (ZX1) was grazing on M. aeruginosa cells. In the control groups, the total MC-LR concentration increased with the growth of M. aeruginosa with an MC-LR content per cell of 0.5–1.5 × 10⁻⁸ μg cell⁻¹. In the treatment with ZX1, the total MC-LR decreased linearly throughout the incubation period. In particular, intracellular MC-LR disappeared with a loss of M. aeruginosa cells in the first few days. Part of the intracellular MC-LR was released to the medium under the grazing stress, resulting in an increase of extracellular MC-LR. The degradation rate of MC-LR was positively related to the initial abundance of ZX1 and negatively related to that of M. aeruginosa. The inhibition ratio of MC-LR production dropped sharply from 98 to 67% when the initial abundance of M. aeruginosa increased from 10⁶ to 10⁷ cells ml⁻¹. However, it increased from 84 to 99% when the initial ZX1 abundance increased from 10⁴ to 10⁵ cells ml⁻¹. The effective removal of both M. aeruginosa cells and MC-LR was observed under lower M. aeruginosa abundance (<10⁶ cells ml⁻¹) and higher ZX1 abundance (>1% of M. aeruginosa abundance). Light had little impact on MC-LR degradation, but MC-LR degradation decreased due to the loss of ZX1 after 10 days of darkness. This study showed that the interactions between M. aeruginosa and ZX1 were strongly influenced by their initial abundances.     

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.     

Spectroscopic and Microscopic Characterization of Uranium Biomineralization in Myxococcus xanthus

In this work, synchrotron-based X-ray absorption spectroscopy (XAS) and transmission electron microscopy (TEM) studies were carried out to elucidate at molecular scale the interaction mechanisms of Myxococcus xanthus with uranium at different pH values. Extended X-ray absorption fine structure (EXAFS) spectroscopic measurements showed that there are significant differences in the structural parameters of the U complexes formed by this bacterium at pH 2 and 4.5. At very low acidic pH of 2, the cells accumulated U(VI) as organic phosphate-metal complexes. At pH 4.5, however, the cells of this bacterium precipitated U(VI) as meta-autunite-like phase. TEM analyses demonstrated that at pH 2 the uranium accumulates were located mainly at the cell surface, whereas at pH 4.5 a uranium precipitation occurred on the cell wall and within the extracellular polysaccharides (EPS) characteristic of this bacterium. Dead/live staining studies showed that 30% and 50% of the uranium treated cell populations were alive at pH 2 and 4.5, respectively. The precipitation of U(VI) as mineral meta-autunite-like phase is possibly due to the bacterial acidic phosphatase activity. The precipitation of uranium as mineral phases may lead to more stable U(VI) sequestration that may be suitable for remediation purposes. These observations, combined with the very high uranium accumulation capability of the studied bacterial cells indicate that M. xanthus may significantly influence the fate of uranium in soil environments where these bacterial species are mainly found.     

Wheat Bran Enhances the Cytotoxicity of Immobilized Alcaligenes aquatilis F8 against Microcystis aeruginosa

Algicidal bacteria offer a promising option for killing cyanobacteria. Therefore, a new Alcaligenes aquatilis strain F8 was isolated to control Microcystis aeruginosa in this study. The algicidal activity of strain F8 was dependent on the cell density of M. aeruginosa, and the maximal algicidal rate of the free bacterium reached 88.45% within 72 h. With a view to its application to the control of M. aeruginosa in the natural environment, strain F8 was immobilized in sodium alginate beads, but immobilization of the strain decreased its algicidal rate compared to that of the free bacterium. However, addition of wheat bran to the sodium alginate matrix used to immobilize strain F8 not only eliminated the adverse effects of immobilization on the bacteria but also resulted in an 8.83% higher algicidal rate of the immobilized than free bacteria. Exclusion and recovery methods were used to identify key ingredients of wheat bran and gain insight into the mechanism underlying the observed enhancement of algicidal activity. This analysis indicated that certain factors in wheat bran, including vitamins B₁, B₂, B₉, and E were responsible for promoting bacterial growth and thereby improving the algicidal rate of immobilized strain F8. Our findings indicate that wheat bran is able to improve the algicidal efficiency of A. aquatilis strain F8 for killing M. aeruginosa and is a good source of not only carbon and nitrogen but also vitamins for bacteria.     

Phytoremediation through Bidens pilosa L., a nonhazardous approach for uranium remediation of contaminated water

Abstract

A few plant species are recognized for uranium bioaccumulation, particularly as upper accumulator. Uranium has a dynamic impact on the physiological, biochemical, and photochemical reactions. Therefore, the superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), soluble sugar, protein, photochemical reactions, and accumulation of uranium characteristics were studied in Bidens pilosa L. while applying altered concentrations of uranium in the form of C₄H₆O₆U. It was done to measure the capacity of B. pilosa L. to remediate uranium from wastewater. In this study, the results showed that B. pilosa L. not only has the ability of uranium accumulation but it can accumulate in the upper parts i.e. leaves and stem. Overall it can accumulate as much as 1538?mg/kg on a dry weight basis. Uranium accumulation is a complex process which changes both physiological and biochemical index in plant species under different treatments. SOD decreased in leaves and stem in response to all treatments whereas POD and CAT increased at treatment 3 and 72?h up to 1335?μ/g-FW and 47?μ/g-FW at 72?h, respectively. This increase was followed by a downward trend. The correlation coefficient between fluorescence ratio F_v/F_m and the concentrations of uranium treatment were significantly negative i.e. ?0.928. The Fourier transform infrared spectroscopy (FTIR) analysis also highlighted that uranium does not change the basic chemical composition of B. pilosa L., but has an effect on the contents of chemical constituents. From the study, it is concluded that B. Pilosa L. has shown a capacity for uranium enrichment, especially as an upper accumulator.     

The effects of K+ growth conditions on the accumulation of cesium by the bacterium Thermus sp. TibetanG6

The accumulation of cesium by the bacterium Thermus sp. TibetanG6 was examined under different K⁺ growth conditions. The effects of external pH and Na⁺ on the accumulation of cesium were also studied, and the mechanism involved was discussed. K⁺ regimes played an important role in the accumulation of cesium by the strain TibetanG6. The quantity of cesium accumulated (24 h) was much higher in K⁺-deficient regime than that in K⁺-sufficient regime. The pH and Na⁺ had different effects on the accumulation of cesium in the two K⁺ regimes. IR spectra analyses indicated that the biosorption is a process of homeostasis with cesium initially accumulated on the cell wall.     

The effects of K+ growth conditions on the accumulation of cesium by the bacterium Thermus sp. TibetanG6

The accumulation of cesium by the bacterium Thermus sp. TibetanG6 was examined under different K⁺ growth conditions. The effects of external pH and Na⁺ on the accumulation of cesium were also studied, and the mechanism involved was discussed. K⁺ regimes played an important role in the accumulation of cesium by the strain TibetanG6. The quantity of cesium accumulated (24 h) was much higher in K⁺-deficient regime than that in K⁺-sufficient regime. The pH and Na⁺ had different effects on the accumulation of cesium in the two K⁺ regimes. IR spectra analyses indicated that the biosorption is a process of homeostasis with cesium initially accumulated on the cell wall.     

Natural uranium impairs the differentiation and the resorbing function of osteoclasts

BackgroundUranium is a naturally occurring radionuclide ubiquitously present in the environment. The skeleton is the main site of uranium long-term accumulation. While it has been shown that natural uranium is able to perturb bone metabolism through its chemical toxicity, its impact on bone resorption by osteoclasts has been poorly explored. Here, we examined for the first time in vitro effects of natural uranium on osteoclasts.MethodsThe effects of uranium on the RAW 264.7 monocyte/macrophage mouse cell line and primary murine osteoclastic cells were characterized by biochemical, molecular and functional analyses.ResultsWe observed a cytotoxicity effect of uranium on osteoclast precursors. Uranium concentrations in the μM range are able to inhibit osteoclast formation, mature osteoclast survival and mineral resorption but don't affect the expression of the osteoclast gene markers Nfatc1, Dc-stamp, Ctsk, Acp5, Atp6v0a3 or Atp6v0d2 in RAW 274.7 cells. Instead, we observed that uranium induces a dose-dependent accumulation of SQSTM1/p62 during osteoclastogenesis.ConclusionsWe show here that uranium impairs osteoclast formation and function in vitro. The decrease in available precursor cells, as well as the reduced viability of mature osteoclasts appears to account for these effects of uranium. The SQSTM1/p62 level increase observed in response to uranium exposure is of particular interest since this protein is a known regulator of osteoclast formation. A tempting hypothesis discussed herein is that SQSTM1/p62 dysregulation contributes to uranium effects on osteoclastogenesis.General significanceWe describe cellular and molecular effects of uranium that potentially affect bone homeostasis.     

Biosorption of uranium and lead by Streptomyces longwoodensis

Biosorption of uranium and lead by lyophilized cells of Streptomyces longwoodensis was examined as a function of metal concentration, pH, cell concentration, and culture age. Cells harvested from the stationary growth phase exhibited an exceptionally high capacity for uranium (0.44 g U/g dry weight) at pH 5. Calculated values of the distribution coefficient and separation factor indicated a strong preference of the cell mass for uranyl ions over lead ions. The specific uranium uptake was similar for the cell wall and the cytoplasmic fraction. Uranium uptake was associated with an increase in hydrogen ion concentration, and phosphorus analysis of whole cells indicated a simple stoichiometric ratio between uranium uptake and phosphorus content. It is proposed that metal ions are bound to phosphodiester residues present both in the cell wall and cytoplasmic fractions. Based on this model, it was shown that uranium accumulation exhibits a maximum at pH 4.6 that is supported by experimental data from previous investigations.     

Quantitative studies on phosphorus transference occuring between <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and its attached bacterium (<Emphasis Type="Italic">Pseudomonas</Emphasis> sp.)

Phosphorus release from Microcystis aeruginosa and attached bacterium (Pseudomonas sp.) isolated from Lake Taihu was examined using a phosphorus isotope tracer in order to investigate the phosphorus transference between the two species. Our results reveal that the amount of phosphorus released form ³²P-saturated M. aeruginosa is determined by its growth phase and most of phosphorus is assimilated by Pseudomonas finally while the amount of phosphorus released from ³²P-saturated Pseudomonas is also determined by the growth phase of M. aeruginosa and most of them are assimilated by M. aeruginosa. The results suggest that phosphorus transference occurs between M. aeruginosa and its attached Pseudomonas . This process makes microenvironment of mucilage of M. aeruginosa attached bacteria maintain relative high amounts of phosphorus. Attached bacteria may be a temporary phosphorus bank to the growth of M. aeruginosa, and assimilation of phosphorus by M. aeruginosa becomes easy when M. aeruginosa is in lag growth phase. Thus, the phosphorus exchange between M. aeruginosa and attached Pseudomonas in microenvironment may be important to microfood web and cyanobacteria bloom.