Sufia K. Kazy

Metal adsorption and desorption by lyophilized Pseudomonas aeruginosa

Biosorption of nickel (Ni2+) and copper (Cu2+) by lyophilized Pseudomonas aeruginosa cells was investigated based on Freundlich isotherm. Bacterial biomass showed significant sorption of both Ni (265 mg g−1) or Cu (137.6 mg g−1), and was also superior over the cation exchanger, IRA 400 (98 mg Ni g−1 or 26.6 mg Cu g−1). Metal binding by the test organism was a fast saturating, pH-dependent process. The optimum pH for Cu adsorption was 7.0 and for Ni 8.0. X-ray diffraction studies revealed that both cations were deposited on the cell predominantly as phosphide crystals. The participation of carboxyl, carbonyl, and phosphoryl groups along with H-bonding in metal sorption was evident in IR spectra. Biomass pretreatment by agents like NaOH, NH4OH or toluene enhanced the metal loading capacity, whereas, oven heating (80°C), autoclaving (120°C, 15 lb (in.2)−1), acid, detergent and acetone treatments were inhibitory. In bimetallic combination, Na, K or Ca increased sorption of Ni as well as Cu in contrast to Cd or Pb. Mineral acids (HCl, H2SO4 and HNO3) and NTA could recover more than 75% (on average) Ni or Cu adsorbed on the biomass. Calcium carbonate (10 mM) was efficient in Ni desorption (71%) compared to Cu (57%). Noticeably sodium carbonate remained specific for Cu remobilization (88%) than Ni (21%). The data are in favour of deployment of the test biomass as an efficient metal removal/recovery system.

Uranium and thorium sequestration by a Pseudomonas sp.: Mechanism and chemical characterization

The mechanism and chemical nature of uranium and thorium sequestration by a Pseudomonas strain was investigated by transmission electron microscopy, energy dispersive X-ray (EDX) analysis, FTIR spectroscopy and X-ray diffractometry. Atomic force microscopy (AFM) used in the tapping mode elucidated the morphological changes in bacterial cells following uranium and thorium binding. Transmission electron microscopy revealed intracellular sequestration of uranium and thorium throughout the cell cytoplasm with electron dense microprecipitations of accumulated metals. Energy dispersive X-ray analysis confirmed the cellular deposition of uranium and thorium. EDX and elemental analysis of sorption solution indicated the binding of uranium and thorium by the bacterial biomass via displacement of cellular potassium and calcium. The strong involvement of cellular phosphate, carboxyl and amide groups in radionuclide binding was ascertained by FTIR spectroscopy. X-ray powder diffraction (XRD) analyses confirmed cellular sequestration of crystalline uranium and thorium phosphates. Overall results indicate that a combined ion-exchange-complexation-microprecipitation mechanism could be involved in uranium and thorium sequestration by this bacterium. Atomic force microscopy and topography analysis revealed an undamaged cell surface with an increase in cell length, width and height following radionuclide accumulation. The arithmetic average roughness (R(a)) and root mean square (RMS) roughness (R(q)) values indicated an increase in surface roughness following uranium and thorium sequestration.

Extracellular polysaccharides of a copper-sensitive and a copper-resistant Pseudomonas aeruginosa strain: synthesis, chemical nature and copper binding

Extracellular polysaccharides (EPS) of a copper-sensitive (Cus) and a copper-resistant (Cur) Pseudomonas aeruginosa strain were investigated in terms of their production, chemical nature and response towards copper exposure. The extent of EPS synthesis by the resistant strain (4.78 mg mg−1 cell dry wt.) was considerably higher over its sensitive counterpart (2.78 mg mg−1 dry wt.). FTIR-spectroscopy and gas chromatography revealed that both the polymers were acidic in nature, containing alginate as the major component along with various neutral- and amino-sugars. Acid content in the Cur EPS (480.54 mg g−1) was greater than that in the Cus EPS (442.0 mg g−1). Presence of Cu2+ in the growth medium caused a dramatic stimulation (approximately 4-fold) in EPS synthesis by the Cur strain, while in a similar condition, the Cus failed to exhibit such response. The polymer of the resistant strain showed elevated Cu2+ binding (320 mg g−1 EPS) compared to that of the sensitive type (270 mg g−1). The overall observations show the potential of the Cur EPS for its deployment in metal bioremediation.

Lanthanum biosorption by a Pseudomonas sp.: equilibrium studies and chemical characterization

Lanthanum biosorption by a Pseudomonas sp. was characterized in terms of equilibrium metal loading, model fitting, kinetics, effect of solution pH, lanthanum–bacteria interaction mechanism and recovery of sorbed metal. Lanthanum sorption by the bacterium was rapid and optimum at pH 5.0 with equilibrium metal loading as high as 950xa0mgxa0g−1xa0biomassxa0dryxa0wt. Scatchard model and potentiometric titration suggested the presence of at least two types of metal-binding sites, corresponding to a strong and a weak binding affinity. The chemical nature of metal–microbe interaction has been elucidated employing FTIR spectroscopy, energy dispersive X-ray analysis (EDX) and X-ray diffraction analysis (XRD). FTIR spectroscopy and XRD analysis revealed strong involvement of cellular carboxyl and phosphate groups in lanthanum binding by the bacterial biomass. EDX and the elemental analysis of the sorption solution ascertained the binding of lanthanum with the bacterial biomass via displacement of cellular potassium and calcium. Transmission electron microscopy exhibited La accumulation throughout the bacterial cell with some granular deposits in cell periphery and in cytoplasm. XRD confirmed the presence of LaPO4 crystals onto the bacterial biomass after La accumulation for a long period. A combined ion-exchange–complexation–microprecipitation mechanism could be involved in lanthanum accumulation by the biomass. Almost 98% of biomass-bound La could be recovered using CaCO3 as the desorbing agent.

Intracellular nickel accumulation by Pseudomonas aeruginosa and its chemical nature

Aims: To investigate intracellular localization of nickel and its chemical nature in Pseudomonas aeruginosa.

Copper uptake and its compartmentalization in Pseudomonas aeruginosa strains: Chemical nature of cellular metal

Copper-sensitive (Cus) and copper-resistant (Cur) strains of Pseudomonas aeruginosa were characterized in terms of Cu2+ sensitivity, uptake and its compartmentalization in the possible cell sectors. Minimum inhibitory concentrations (MICs) of Cu2+ for the Cur strain (3.2 mM and 0.12 mM in enriched- and in minimal-medium, respectively) were almost 5-fold higher over that of its sensitive counterpart. While Cus strain accumulated Cu2+ to a maximum of 1.8 μ mol mg−1 protein, Cur strain increased it to 2.37 μmol mg−1 protein. Both the strains also demonstrated energy- and pH-dependent Cu2+ uptake through the broad-substrate range divalent cation (Zn2+, Mg2+, Co2+) uptake system as well as through the system specific for Cu2+. Cell-fractionation study revealed that in Cur strain, periplasm and membrane are the main Cu2+ binding sites, whereas, in case of Cus strain, it is the cytoplasm. The overall observations indicate that the Cur strain restricted Cu2+ sequestration exterior to the cytoplasm as the possible strategy for Cu-resistance. The chemical nature of Cu2+ deposition in the respective strains was also ascertained by X-ray powder diffraction analysis.

Uranium Sorption by Pseudomonas Biomass Immobilized in Radiation Polymerized Polyacrylamide Bio-Beads

A Pseudomonas strain identified as a potent biosorbent of uranium (U) and thorium was immobilized in radiation-induced polyacrylamide matrix for its application in radionuclide containing wastewater treatment. The immobilized biomass exhibited a high U sorption of 202 mg g−1 dry wt. with its optimum at pH 5.0. A good fit of experimental data to the Freundlich model suggested multilayered uranium binding with an affinity distribution among biomass metal binding sites. Scanning electron microscopy revealed a highly porous nature of the radiation-polymerized beads with bacterial cells mostly entrapped on pore walls. Energy dispersive X-ray analysis (EDXA) coupled with SEM ascertained the accumulation of uranium by the immobilized biomass without any physical damage to the cells. A significant (90%) part of biosorbed uranium was recovered using sodium bicarbonate with the immobilized biomass maintaining their U resorption capacity for multiple sorption–desorption cycles. Uranium loading and elution behavior of immobilized biomass evaluated within a continuous up-flow packed bed columnar reactor showed its effectiveness in removing uranium from low concentration (50 mg U L−1) followed by its recovery resulting in a 4–5-fold waste volume reduction. The data suggested the suitability of radiation polymerization in obtaining bacterial beads for metal removal and also the potential of Pseudomonas biomass in treatment of radionuclide containing waste streams.

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

ABSTRACT 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.

Nickel uptake by Pseudomonas aeruginosa : Role of modifying factors

Abstract.Pseudomonas aeruginosa cells growing in minimal medium were 40-fold more sensitive to Ni2+ than cells growing in enriched medium, suggesting a possible protective role of medium ingredients. Likewise, cells pre-grown in enriched medium showed a high Km (6.15 mM) and increased Ni2+ uptake (950 nmol mg−1 protein, 1h) over cells pre-sown in minimal medium (Km, 0.48 mM; 146 nmol mg−1 protein, 1 h). The overall pattern indicates that cells pre-grown in enriched medium were characterized by having lowered affinity towards Ni2+ than those with minimal medium background. The enhanced Ni2+ uptake by enriched medium-grown cells can be correlated with the improved metabolic state of the cells. Ni2+ uptake was optimum at neutrality (pH 7.0). A major Ni2+ transport system was competitively inhibited by Mg2+, Zn2+, Cd2+, or Co2+ (400 μM each). Noticeably, a minor Ni2+ transport pathway was still operative even in the higher concentration range of Mg2+ (4 mM and 40 mM). The stimulation of Ni2+ uptake monitored in the presence of different carbon sources (0.5% wt/vol, each) showed the sequence: glucose (1.6-fold) > phenol = gallic acid (1.5-fold). Succinate, in comparison, reduced Ni2+ uptake (0.5-fold) possibly because of its acting as a metal chelator as well. Sensitivity of Ni2+ transport towards methyl viologen, azide, 2-4 DNP, and DCCD suggested that transport was energy-linked.

Molecular assessment of microbial diversity and community structure at uranium mines of Jaduguda, India

Microbial diversity associated with uranium mine areas of Jaduguda, India has been investigated using a culture independent molecular approach. Soil samples collected from existing and proposed mine sites were analyzed for physicochemical parameters. Community DNA was extracted from five samples. Small subunit rRNA gene (16S rRNA) was PCR amplified using bacterial primers. The diversity of the total bacterial community was described at molecular level by amplified ribosomal DNA restriction analysis (ARDRA). Dominant bacterial groups (represents by OTUs) selected by ARDRA were identified by sequencing the 16S rRNA genes. From the bacterial rDNA clone library around 230 clones were used for further analysis. The unique OTUs and number of clones representing such OTUs were determined. Dominant OTUs were sequenced and identified. These phylotypes spanned a wide range within the bacterial domain occupying Proteobacteria, Acidobacteria, Bacteroidetes, Firmicutes, Cyanobacteria as major phyla. About 46 % of clones sequenced from various sites were identified as Proteobacteria. The present findings on microbial diversity at the molecular level are the first of its kind for uranium mine sites of India. Around 20 % of the clone sequences showed little affiliation with known taxa and probably represent new organisms adapted to this habitat.