Ana Marqués

URANIUM ACCUMULATION BY PSEUDOMONAS SP. EPS-5028

SummaryPseudomonas sp. EPS-5028 was examined for the ability to accumulate uranium from solutions. The uptake of uranium by this microorganism is very rapid and is affected by pH but not by temperature, metabolic inhibitors, culture time and the presence of various cations and anions. The amount of uranium absorbed by the cells increased as the uranium concentration of the solution increased up to 55 mg uranium/g cell dry weight. Electron microscopy indicated that uranium accumulated intracellularly as needle-like fibrils. Uranium could be removed chemically from the cells, which could then be reused as a biosorbent.

Effect of pH on the biosorption of nickel and other heavy metals by Pseudomonas fluorescens 4F39

Accumulation of heavy metals by Pseudomonas fluorescens 4F39 was rapid and pH-dependent. The affinity series for bacterial accumulation of metal cations decreased in the order Ni>>Hg>U>>As>Cu>Cd>Co>Cr>Pb. Metal cations were grouped into those whose accumulation increased as the pH increased, with a maximum accumulation at the pH before precipitation (Ni, Cu, Pb, Cd, Co), and those whose maximum accumulation was not associated with precipitation (Cr, As, U, Hg). High Ni2+ accumulation was studied. Electron microscopy indicated that at pH 9, Ni2+ accumulated on the cell surface as needle and hexagon-like precipitates, whose crystalline structure was confirmed by electron diffraction analysis and corresponded to two different orientations of the nickel hydroxide crystals. Crystals on cells showed marked anisotropy by X-ray powder diffraction, which differentiated them from crystals observed in nickel solution at pH 10 and 11 and from commercial nickel hydroxide. Nickel biosorption by Pseudomonas fluorescens 4F39 was a microprecipitation consequence of an ion exchange. Journal of Industrial Microbiology & Biotechnology (2000) 24, 146–151.

Nickel biosorption by free and immobilized cells of Pseudomonas fluorescens 4F39: A comparative study

The biosorption of nickel ions on Pseudomonasfluorescens 4F39 free cells or immobilized cells in beads of agar (biobeads)has been studied in batch experiments to determine the effect ofcell immobilization on the metal accumulation properties of bothsystems. Bacterial cells were immobilized in agar beads followingthe interphase technique. When free cells were used, the sorptionequilibrium was reached in 5 min but with biobeads it took 24 hr as a consequence of metal diffusion. The pH of the Ni2+solution was found to be critical for Ni2+ accumulation,the optimum being 8, although the magnitude of this effect waslower in immobilized cells. The equilibrium data have been analysed using the Langmuir adsorption model. The qmax of free cells, immobilized cells and biobeads was 145, 37 and7.6 mg Ni2+/g dry sorbent, respectively. The removal capacity of free cells and immobilized cells increased when the cell concentration decreased. The maximum removal efficiency ofbiobeads was obtained when the cell concentration was 1.43 mg drycells/mL Ni2+ solution. The agar concentration in biobeads affected the Ni2+ accumulation, the optimum being 2%. Desorption of Ni2+ with 0.5 mM dipicolinic acid was efficient. Cycles of accumulation/desorption resulted in a lossof non immobilized cells. An increase of the removal efficiencyfrom the first cycle of accumulation/desorption was observed with biobeads.

The interphase technique: a simple method of cell immobilization in gel-beads

A variation of the emulsion techniques for microbial cell immobilization by encapsulation in gel beads is described. The utilized gels are silica gel or natural polymers like agar, agarose, phytagel or carrageenan. This technique utilizes the interfacial tension that is formed between two liquids of different polarity. The gel-beads are easily obtained and homogeneous in size.

Heavy metal biosorption by gellan gum gel beads

The Ni(2+) accumulation in batch mode from diluted solutions by gel beads of gellan gum (GG), alginate, kappa-carrageenan, agar, agarose, silica gel, polyacrylamide and two mixtures of GG+agar was investigated. All polymeric materials studied accumulated Ni(2+), but gel beads of GG were stable, easily obtainable and showed the highest Ni(2+) accumulation. The pH of the Ni(2+) solution was not critical for Ni(2+) accumulation. Accumulation of metals Cu(2+), Co(2+), Ni(2+), Pb(2+), Cd(2+) and Zn(2+) by GG gel beads reached the equilibrium after 24h. The removal of Pb(2+) and Cu(2+) from the aqueous solution was very efficient, with maximum metal uptake (q(max)) of 0.85 and 0.75 mmol/g dw GG, respectively. The general q(max) sequence was Pb>Cu>Ni approximately Zn=Co>Cd. In an equimolar metal mixture sorption experiment a clear reduction in accumulation was observed, except for Pb(2+) (30%). Heavy metals were desorbed with 100mM sodium citrate.

Permeabilization of biological and artificial membranes by a bacterial dirhamnolipid produced by Pseudomonas aeruginosa.

Pseudomonas aeruginosa, when cultured under the appropriate conditions, secretes rhamnolipids to the external medium. These glycolipids constitute one of the most interesting classes of biosurfactants so far. A dirhamnolipid fraction was isolated and purified from the crude biosurfactant, and its action on model and biological membranes was studied. Dirhamnolipid induced leakage of internal contents, as measured by the release of carboxyfluorescein, in phosphatidylcholine unilamellar vesicles, at concentrations below its CMC. Membrane solubilization was not observed within this concentration range. The presence of inverted cone-shaped lipids in the membrane, namely lysophosphatidylcholine, accelerated leakage, whereas cone-shaped lipids, like phosphatidylethanolamine, decreased leakage rate. Increasing concentrations of cholesterol protected the membrane against dirhamnolipid-induced leakage, which was totally abolished by the presence of 50 mol% of the sterol. Dirhamnolipid caused hemolysis of human erythrocytes through a lytic mechanism, as shown by the similar rates of K(+) and hemoglobin leakage, and by the absence of effect of osmotic protectants. Scanning electron microscopy showed that the addition of the biosurfactant changed the usual disc shape of erythrocytes into that of spheroechinocytes. The results are discussed within the frame of the biological actions of dirhamnolipid, and the possible future applications of this biosurfactant.

The physicochemical properties and chemical composition of trehalose lipids produced by Rhodococcus erythropolis 51T7

This study analyzed the chemical and physical properties of a biosurfactant synthesized by Rhodococcus sp. 51T7. The biosurfactant was a trehalose tetraester (THL) consisting of six components: one major and five minor. The hydrophobic moieties ranged in size from 9 to 11 carbons. The critical micelle concentration (CMC) was 0.037g L(-1) and the interfacial tension against hexadecane was 5mN m(-1). At pH 7.4 the glycolipid CMC/critical aggregation concentration (CAC) was 0.05g L(-1) and at pH 4 it was 0.034g L(-1). A phase diagram revealed effective emulsification with water and paraffin or isopropyl myristate. A composition of 11.3-7.5-81.8 (isopropyl myristate-THL-W) was stable for at least 3 months. The HLB was 11 and the phase behaviour of the glycolipid revealed the formation of lamellar and hexagonal liquid-crystalline textures.

Removal of uranium by an exopolysaccharide from Pseudomonas sp.

SummaryAccumulation of uranium (U) is reported for isolated exopolysaccharide produced by Pseudomonas sp. EPS-5028. A maximum uptake of 96 μg U/mg polymer was observed. In contrast, the maximum accumulation of uranium by deacylated polysaccharide was 46 μg/mg. This metal-complexing capacity observed suggests that the anionic reactive sites on the structure could be responsible for this activity. Metal uptake was affected by pH and was not affected by temperature. Expolysaccharide from Pseudomonas sp. EPS-5028 obeyed the Freundlich isotherm indicating single layer adsorption.

Isolation and partial characterization of a biosurfactant mixture produced by Sphingobacterium sp. isolated from soil

Strain 6.2S, isolated from soil and identified as a Sphingobacterium sp., is the first strain in this genus to be reported as a biosurfactant producer, being able to reduce the surface tension of its culture supernatant to 32 mN/m. In this work, biosurfactants from the culture supernatant were purified and partially characterized. The crude extract (10 g/L) was very effective in reducing surface tension (22 mN/m). Thin layer chromatography (TLC) indicated that a mixture of various biosurfactants was present in the 6.2S crude extract. After purification, Fraction A, a phospholipid mixture, reduced surface tension to 33 mN/m. Fraction B was a mixture of lipopetides and at least one glycolipid. The surface tension-concentration curve showed two plateaux, the first of which can be attributed to a critical aggregation concentration of the biosurfactant with a protein (2.7 g/L) and the second to the true cmc in water (6.3g/L).

Interactions of a bacterial biosurfactant trehalose lipid with phosphatidylserine membranes

Trehalose lipids are biosurfactants produced by rhodococci that, in addition to their well known potential industrial and environmental uses, are gaining interest in their use as therapeutic agents. The study of the interaction of biosurfactants with membranes is important in order to understand the molecular mechanism of their biological actions. In this work we look into the interactions of a bacterial trehalose lipid produced by Rhodococcus sp. with dimyristoylphosphatidylserine membranes by using differential scanning calorimetry, X-ray diffraction and infrared spectroscopy. Differential scanning calorimetry and X-ray diffraction show that trehalose lipid broadens and shifts the phospholipid gel to liquid-crystalline phase transition to lower temperatures, does not modify the macroscopic bilayer organization and presents good miscibility both in the gel and the liquid-crystalline phases. Infrared experiments show that trehalose lipid increases the fluidity of the phosphatidylserine acyl chains, changed the local environment of the polar head group, and decreased the hydration of the interfacial region of the bilayer. Trehalose lipid was also able to affect the thermotropic transition of dimyristoylphosphatidyserine in the presence of calcium. These results support the idea that trehalose lipid incorporates into the phosphatidylserine bilayers and produces structural perturbations which might affect the function of the membrane.