Biosorption of Lead,Copper, and Cadmium by <Emphasis Type="Italic">Phanerochaete chrysosporium</Emphasis> in Ternary Metal Mixtures: Statistical Analysis of Individual and Interaction Effects

Biosorption of three divalent metals, viz., lead, copper, and cadmium in ternary aqueous mixtures was studied using Phanerochaete chrysosporium in batch shake flasks. The mixtures were prepared containing the metals at their either varying optimum or equal initial concentration combinations in aqueous solution of pH optimum to each of the metals. Following were the optimum initial concentration ranges of the metals in mixture: lead, 60–100 mg/L; copper, 20–60 mg/L; and cadmium, 5–15 mg/L. And, for varying these optimum concentration levels of the metals, a 2³ full factorial design of experiments was employed. The results revealed that an increase in lead and cadmium concentrations helped in their better biosorption by the fungus, but an increase in initial copper concentration slightly diminished its removal. Statistical analysis of the results in the form of analysis of variance and Student t test gave a clear interpretation on the roles of both the individual metals and their interactions in the uptake of metals from mixture. Compared to the uptake of metals when presented individually, lead biosorption in mixture was found to be enhanced to a degree as high as 99%; on the other hand, copper and cadmium removals from mixtures were inhibited to the extent of 100% and 98%, respectively. However, this extent of inhibition or enhancement in the metal removals compared to the individual removals was less in mixtures containing all equal concentrations of the metals.     

Removal and recovery of copper (II) ions by bacterial biosorption

Studies were conducted toinvestigate the removal and recovery of copper (II) ions from aqueous solutions by Micrococcus sp., which was isolated from a local activated sludge process. The equilibrium of copper biosorption followed the Langmuir isotherm model very well with a maximum biosorption capacity (q_max) of 36.5 mg of Cu²⁺/gofdry cell at pH 5.0 and 52.1 mg of Cu²⁺/g of dry cell at pH 6.0. Cells harvested at exponential growth phase and stationary phase showed similar biosorption characteristics for copper, Copper uptake by cells was negligible at pH 2.0 and then increased rapidly with increasing pH un til 6.0. In multim etal systems, Micrococcus sp. exhibited a preferential biosorption order: Cu−Pb>Ni−Zn. There is virtually no interference with copper uptake by Micrococcus sp. from solutions bearing high concentrations of Cl⁻, SO ₄ ²⁻ , and NO3/− (0–500 mg/L). Sulfuric acid (0.05 M) was the most efficient desorption medium, recovering >90% of the initial copper sorbed. The copper capacity of Micrococcus sp. remained unchanged after five successive sorption and desorption cycles. Immobilization of Micrococcus sp. in 2% calcium alginate and 10% polyacrylamide gel beads increased copper uptake by 61%. Biomass of Micrococcus sp. may be applicable to the development of potentially cost-effective biosorbent for removing and recovering copper from effluents.     

Study on the characterization of lead (II) biosorption by fungus <Emphasis Type="Italic">Aspergillus parasiticus</Emphasis>

The lead (II) biosorption potential of Aspergillus parasiticus fungal biomass has been investigated in a batch system. The initial pH, biosorbent dosage, contact time, initial metal ion concentrations and temperature were studied to optimize the biosorption conditions. The maximum lead (II) biosorption capacity of the fungal biosorbent was found as 4.02 × 10⁻⁴ mol g⁻¹ at pH 5.0 and 20°C. The biosorption equilibrium was reached in 70 min. Equilibrium biosorption data were followed by the Langmuir, Freundlich and Dubinin-Radushkevich (D-R) isotherm models. In regeneration experiments, no significant loss of sorption performance was observed during four biosorption-desorption cycles. The interactions between lead (II) ions and biosorbent were also examined by FTIR and EDAX analysis. The results revealed that biosorption process could be described by ion exchange as dominant mechanism as well as complexation for this biosorbent. The ion exchange mechanism was confirmed by E value obtained from D-R isotherm model as well.     

Efficient uranium(VI) biosorption on grapefruit peel: kinetic study and thermodynamic parameters

The uranium(VI) biosorption by grapefruit peel was studied from aqueous solutions. Batch experiments was conducted to evaluate the effect of contact time, initial uranium(VI) concentration, initial pH, adsorbent dose, salt concentration and temperature. The equilibrium process was well described by the Langmuir, Redlich–Peterson and Koble–Corrigan isotherm models, with maximum sorption capacity of 140.79 mg g⁻¹ at 298 K. The pseudo second order model and Elovish model adequately describe the kinetic data in comparison to the pseudo first order model and the process involving rate-controlling step is much complex involving both boundary layer and intra-particle diffusion processes. The effective diffusion parameter D _i and D _f values were estimated at different initial concentration and the average values were determined to be 1.167 × 10⁻⁷ and 4.078 × 10⁻⁸ cm² s⁻¹. Thermodynamic parameters showed that the biosorption of uranium(VI) onto grapefruit peel biomass was feasible, spontaneous and endothermic under studied conditions. The physical and chemical properties of the adsorbent were determined by SEM, TG-DSC, XRD and elemental analysis and the nature of biomass–uranium (VI) interactions was evaluated by FTIR analysis, which showed the participation of COOH, OH and NH₂ groups in the biosorption process. Adsorbents could be regenerated using 0.05 mol L⁻¹ HCl solution at least three cycles, with up to 80% recovery. Thus, the biomass used in this work proved to be effective materials for the treatment of uranium (VI) bearing aqueous solutions.     

Biosorption of heavy metals by bacteria isolated from activated sludge

Twelve aerobic bacteria from activated sludge were isolated and identified. These included both Gram-positive (e.g., Bacillus) and Gram-negative (e.g., Pseudomonas) bacteria. The biosorption capacity of these strains for three different heavy metals (copper, nickel, and lead) was determined at pH 5.0 and initial metal concentration of 100 mg/L. Among these 12 isolates, Pseudomonas pseudoalcaligenes was selected for further investigation owing to its high metal biosorption capacity. The lead and copper biosorption of this strain followed the Langmuir isotherm model quite well with maximum biosorption capacity (q _max) reaching 271.7mg of Pb²⁺/g of dry cell and 46.8 mg of Cu²⁺/g of dry cell at pH 5.0. Study of the effect of pH on lead and copper removal indicated that the metal biosorption increased with increasing pH from 2.0 to 7.0. A mutual inhibitory effect was observed in the lead-copper system because the presence of either ion affected the sorption capacity of the other. Unequal inhibitions were observed in all the nickel binary systems. The increasing order of affinity of the three metals toward P. pseudoalcaligenes was Ni<Cu<Pb. The metal biosorptive potential of these isolates, especially P. pseudoalcaligenes, may have possible applications in the removal and recovery of metals from industrial effluents.     

Determination of Ultratrace Amounts of Copper and Cadmium in Seawater by Graphite Furnace Atomic Absorption Spectrometry with Flow-Injection Semi On-line Preconcentration

Methods are described for the determination of ultratrace amounts of copper and cadmium in seawater by graphite furnace atomic absorption spectrometry with flow-injection, microcolumn preconcentration. A new type of C₁₈ column loaded with sodium diethyldithiocarbamate (sodium-DDC) was used to extract copper and cadmium from seawater as the DDC chelates. The analytical effects of the pH of the mixture of the sample and sodium-DDC solutions and the concentration of the chelating reagent were studied. Sodium-DDC-loaded columns and unloaded C₁₈ columns with different shapes and volumes were compared. To determine copper in seawater, a simple aqueous calibration was made with a mixture of palladium and magnesium nitrate as a matrix modifier, while for cadmium no matrix modifier was necessary. This method required only small seawater volumes, 600 and 400 μl for the determination of copper and cadmium respectively, with preconcentration factors of 15-fold for copper and 10-fold for cadmium. Detection limits for the preconcentration of aqueous solutions of copper and cadmium were 0.024 and 0.004 μg liter⁻¹ (3σ), respectively. Results for determinations of copper and cadmium in National Research Council of Canada, CASS-2, Nearshore Seawater Reference Material showed no significant differences between the certified values and the measured values, based on Student′s t test at the 95% confidence level. The relative standard deviations of the various measurements varied between 2 and 8%.     

Sorption behavior of Am<Superscript>3+</Superscript> on suspended pyrite

Sorption behavior of ²⁴¹Am (~10⁻⁹ M) on naturally occurring mineral pyrite (particle size: ≤70 μm) has been studied under varying conditions of pH (2–11), and ionic strength (0.01–1.0 M (NaClO₄)). The effects of humic acid (2 mg/L), other complexing anions (1 × 10⁻⁴ M CO₃ ²⁻, SO₄ ²⁻, C₂O₄ ²⁻ and PO₄ ³⁻), di- and trivalent metal ions (1 × 10⁻³ M Mg²⁺, Ca²⁺ and Nd³⁺) on sorption behavior of Am³⁺ at a fixed ionic strength (I = 0.10 M (NaClO₄)) have been studied. The sorption of ²⁴¹Am on pyrite increased with pH from 2.8 (84%) to 8.1 (97%). The sorption of ²⁴¹Am decreased with ionic strength at low pH values (2 ≤ pH ≤ 4), but was insensitive in the pH range of 4–10, suggesting the formation of outer-sphere complexes on pyrite surface at lower pH, and inner-sphere complexes at higher pH values. The sorption of ²⁴¹Am increased in the presence of (i) humic acid (5 < pH < 7.5), and (ii) C₂O₄ ²⁻ (2 < pH < 3). By contrast, other complexing anions such as (carbonate, phosphate, and sulphate) showed negligible influence on ²⁴¹Am sorption. The presence of Mg²⁺, Ca²⁺ ions showed marginal effect on the sorption profile of ²⁴¹Am; while the presence of Nd³⁺ ion suppressed its sorption significantly under the conditions of present study. The sorption of ²⁴¹Am on pyrite decreased with increased temperature indicating an exothermic process.     

Transport and sorption of <Superscript>85</Superscript>Sr and <Superscript>125</Superscript>I in crushed crystalline rocks under dynamic flow conditions

Transport and sorption of water-soluble ⁸⁵Sr²⁺ and ¹²⁵I⁻ in the columns with beds of crushed crystalline rocks from synthetic groundwater has been studied under dynamic flow conditions. Samples of crystalline rocks: diorite-I, diorite-II, gabbro, granite and tonalite, having the grain size between 0.25 and 0.80 mm, were used. Plastic syringes of 8.8 cm length and 2.1 cm in diameter were applied as columns. The synthetic groundwater was pumped downward through the columns with a seepage velocity of about 0.2 cm/min and the given radioactive nuclide was added into the water stream individually in a form of a short pulse. In case of ⁸⁵Sr, desorption from diorite-I was also studied using an artificial acid rainfall and then, the longitudinal distribution of the residual ⁸⁵Sr activity along the bed was measured. Retardation, distribution and hydrodynamic dispersion coefficients were determined by the evaluation of respective breakthrough curves. A corrected integral form of a simple advection–dispersion equation was derived and used for fitting the experimental data. The K _d-parameters resulting from dynamic experiments were also compared with the results of static sorption experiments.     

Formation and Stability of Cadmium(II)/Phytate Complexes by Different Electrochemical Techniques. Critical Analysis of Results

In the present work the stability constants of various cadmium(II)/phytate (Phy) species were determined at T=298.15 K in NaNO₃(aq) at I=0.1 mol⋅L⁻¹ by DP-ASV (Differential Pulse Anodic Stripping Voltammetry) and by potentiometric titrations using an ISE-Cd²⁺. Cyclic voltammograms were also recorded to check the electrochemical behavior of cadmium in the presence of phytate. The results were analyzed together with previous data determined by ISE-H⁺ measurements. Data obtained were used to provide an exhaustive speciation scheme for the phytate/cadmium(II) system at different conditions, as well as a comprehensive representation of the binding ability of phytate toward cadmium(II). Different pL₅₀ values {a previously proposed empirical parameter, expressed as −log ₁₀ C _Phy, where C _Phy is the total phytate concentration necessary to bind 50% cadmium(II)} were also calculated by modeling its dependence on pH.     

Bioadsorption and ion exchange of Cr3+ and Pb2+ solutions with algae

Heavy metals can be removed from effluents and recovered using physico-chemical mechanisms as biosorption processes. In this work “Arribada” seaweed biomass was employed to assess its biosorptive capacity for the chromium (Cr³⁺) and lead (Pb²⁺) cations that usually are present in waste waters of plating industries. Equilibrium and kinetic experiments were conducted in a mixed reactor on a batch basis. Biosorption equilibrium and fluid-solid mass transfer constants data were analyzed through the concept of ion exchange sorption isotherm. The respective equilibrium exchange constants (K _eqCr=173.42, K _eqPb=58.86) and volumetric mass transfer coefficients ((k _mCr a)′=1.13×10⁻³ s⁻¹, (k _mPb a)′=0.89×10⁻³ s⁻¹) were employed for the dynamic analysis of Cr and Pb sorption in a fixed-bed flow-through sorption column. The breakthrough curves obtained for both metals were compared with the predicted values by the heterogeneous model (K _eqCr=171.29, K _eqPb=60.14; k _mCr a=7.81×10⁻² s⁻¹, k _mPb a=2.43×10⁻² s⁻¹), taking into account the mass transfer process. The results suggest that these algae may be employed in a metal removal/recovery process at low cost. An erratum to this article can be found at     

Determination of Kinetic Parameters in the Biosorption of Cr (VI) on Immobilized <Emphasis Type="Italic">Bacillus cereus</Emphasis> M<Superscript>1</Superscript><Subscript>16</Subscript> in a Continuous Packed Bed Column Reactor

Due to technological advancement, environment suffers from untreated toxic heavy metal bearing effluent coming from different industries. Chromium (VI) is one of those heavy metals having adverse impact on ecological balance, human, and plant health because of its carcinogenic properties. Biosorption is presented as an alternative to traditional technologies which are costly and inefficient for treatment of industrial wastes containing low amount of heavy metals. In this study, bioremediation of Cr (VI) ions by immobilized Bacillus cereus M¹ ₁₆ was investigated in a laboratory scale packed bed up-flow column reactor. The effect of important parameters, such as the inlet flow rate, influent concentration, and effective bed height, has been studied. External mass transfer, surface adsorption, and intrabead mass transfer were also studied to conclude the rate limiting step for removal of Cr (VI) and to determine the process parameters which are important for biosorption optimization. The external mass transfer coefficient was calculated at different flow rates (6.51 × 10⁻² to 7.58 × 10⁻² cm/min). Using the model, the surface adsorption rate constant (k _ad) and the intrabead mass transfer coefficient (k _i) were predicted as 0.0267 × 10⁻³ and 0.7465 × 10⁻³ l/g/min, respectively. Both are much lower than the external mass transfer coefficient (k _e). The surface adsorption phenomenon is acting as the rate-limiting step due to its high resistance for removal of Cr (VI).     

ESR studies of doping C60 fullerene by zinc, cadmium, copper, gallium, and indium vapors

In the ESR spectra recorded at the beginning of the doping reaction of C₆₀ fullerene by zinc, gallium, and indium vapors, signals corresponding to C₆₀ ⁻ were observed. In the course of reduction, these signals disappeared and reversibly transformed into the signals of higher charged C₆₀ ⁿ⁻ anions. In the case of cadmium and copper, only the spectra of C₆₀ ⁿ⁻ were recorded. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 2, pp. 303–305, February, 1997.     

1H-NMR study of Na alginates extracted from Sargassum spp. in relation to metal biosorption

The use of a number of species of marine brown algae in the implementation of bioremediation strategies for toxic heavy metals is being considered and evaluated. The biosorption capacity of these algae for heavy metals resides mainly in a group of linear polysaccharides known as alginates that occur as a gel in the algal thallus. The potential for selective metal binding by the biomass of two species of Sargassum was evaluated by ¹H-NMR (nuclear magnetic resonance) following a high temperature, alkaline extraction and purification of their alginate polysaccharide. The alkaline extraction protocol applied to Sargassum fluitans and Sargassum siliquosum yielded alginate samples of low viscosity, suitable for direct acquisition of well-resolved spectra. Estimates of both the ratio of β-d-mannopyranuronosyl (M) and α-l-gulopyranuronosyl (G) residues along the polymer chain and the frequencies of occurrence of diad uronic acid residue pairs were obtained. Guluronic acid (G) was the major component in all, extracts and the GG diads accounted for more than 49% of the polymer diads. Whereas the performance of Sargassum spp. in the metal biosorption process is a function of both its alginate content and composition, the occurrence of “G-blocks” in both purified alginates and in the raw brown seaweed is critical because it results in a well-established selectivity for divalentions, potentially increasing the commercial effectiveness of targeted biosorption as a means of remediation.     

Profiling of the reactive oxygen species-related ecotoxicity of CuO, ZnO, TiO2, silver and fullerene nanoparticles using a set of recombinant luminescent Escherichia coli strains: differentiating the impact of particles and solubilised metals

We propose a novel combination of high-throughput luminescent bacterial tests for the evaluation of the reactive oxygen species (ROS)-generating potential of engineered nanoparticles (eNPs) and the role of solubilised metal ions in this process. The set of tests consists of differently engineered recombinant Escherichia coli strains: (1) a new sensor strain, which bioluminescence is induced by superoxide anions; (2) six recombinant E. coli strains (superoxide dismutase (sod) single, double and triple mutants and a respective wild-type strain), transformed with luxCDABE genes responding to toxic compounds by decreasing their luminescence; and (3) three strains in which bioluminescence is specifically induced by bioavailable metals (Cu, Zn and Ag). The applicability of this battery of tests in profiling oxidative potential of eNPs was evaluated on nTiO₂, nCuO, nZnO and nAg (25, 30, 70 and <100 nm, respectively) NPs and fullerenes. As controls for the size or solubility, the bulk formulations (bTiO₂, bCuO and bZnO) and soluble salts (ZnSO₄, CuSO₄ and AgNO₃) were also analysed. Bacterial toxicity tests showed that nCuO was four-fold more toxic, and nAg was 15-fold more toxic to triple sod mutant than to wild type (2-h EC₅₀ values were 8.1 and 2.0 mg Cu l⁻¹, respectively, and 46 and 3.1 mg Ag l⁻¹, respectively). Formation of ROS by nCuO and nAg was proved by superoxide anion-inducible strain. The metal sensor bacteria showed that the ROS formation by CuO NPs was caused by solubilised Cu ions, but in case of nAg, particles also had an effect. nZnO was remarkably more toxic to sod triple mutant than to wild type strain (2-h EC₅₀ were 4.5 and 54 mg Zn l⁻¹, respectively). Fullerenes inhibited the bioluminescence of sod triple mutant at 3,882 mg l⁻¹ but had no effect on the wild-type strain even at 20,800 mg l⁻¹. Nano and bTiO₂ showed some effect on viability of bacteria only at high concentrations (>4,000 mg l⁻¹) although nTiO₂ (but not bTiO₂) induced the bioluminescence of the superoxide anion sensing bacteria starting from 100 mg l⁻¹. Thus, our innovative combined approach is expected to provide more consistent and informative data concerning the general toxicity, ROS-production potential and also solubilisation of metals in the case of metallic NPs.     

Sorption of cesium from water solutions on potassium nickel hexacyanoferrate-modified <Emphasis Type="Italic">Agaricus bisporus</Emphasis> mushroom biomass

In order to gain biosorbent that would have the ability to bind cesium ions from water solution effectively, potassium nickel hexacyanoferrate(II) (KNiFC) was incorporated into the mushroom biomass of Agaricus bisporus. Cesium sorption by KNIFC-modified A. bisporus biosorbent was observed in batch system, using radiotracer technique using ¹³⁷Cs radioisotope. Kinetic study showed that the cesium sorption was quite rapid and sorption equilibrium was attained within 1 h. Sorption kinetics of cesium was well described by pseudo-second order kinetics. Sorption equilibrium was the best described by Freundlich isotherm and the distribution coefficient was at interval 7,662–159 cm³ g⁻¹. Cesium sorption depended on initial pH of solution. Cesium sorption was very low at pH₀ 1.0–3.0. At initial pH 11.0, maximum sorption of cesium was found. Negative effect of monovalent (K⁺, Na⁺, NH₄ ⁺) and divalent (Ca²⁺, Mg²⁺) cations on cesium sorption was observed. Desorption experiments showed that 0.1 M potassium chloride is the most suitable desorption agent but the complete desorption of cesium ions from KNiFC-modifed biosorbent was not achieved.     

Scale up and Application of Biosurfactant from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in Enhanced Oil Recovery

There is a lack of fundamental knowledge about the scale up of biosurfactant production. In order to develop suitable technology of commercialization, carrying out tests in shake flasks and bioreactors was essential. A reactor with integrated foam collector was designed for biosurfactant production using Bacillus subtilis isolated from agricultural soil. The yield of biosurfactant on biomass (Y _p/x), biosurfactant on sucrose (Y _p/s), and the volumetric production rate (Y) for shake flask were obtained about 0.45 g g⁻¹, 0.18 g g⁻¹, and 0.03 g l⁻¹ h⁻¹, respectively. The best condition for bioreactor was 300 rpm and 1.5 vvm, giving Y _x/s, Y _p/x, Y _p/s, and Y of 0.42 g g⁻¹, 0.595 g g⁻¹, 0.25 g g⁻¹, and 0.057 g l⁻¹ h⁻¹, respectively. The biosurfactant maximum production, 2.5 g l⁻¹, was reached in 44 h of growth, which was 28% better than the shake flask. The obtained volumetric oxygen transfer coefficient (K _L a) values at optimum conditions in the shake flask and the bioreactor were found to be around 0.01 and 0.0117 s⁻¹, respectively. Comparison of K _L a values at optimum conditions shows that biosurfactant production scaling up from shake flask to bioreactor can be done with K _L a as scale up criterion very accurately. Nearly 8% of original oil in place was recovered using this biosurfactant after water flooding in the sand pack.     

Kinetics and mechanism of interaction of hexaaquachromium(III) with <Emphasis Type="SmallCaps">l</Emphasis>-Dopa in aqueous medium: comparative antiparkinsonian studies

The kinetics and mechanism of the substitution reaction between [Cr(H₂O)₆]³⁺ and l-Dopa in aqueous medium has been studied over the range 1.8 ≤ pH ≤ 2.6, 1.68 × 10⁻² mol dm⁻³ ≤ [Dopa] ≤ 5.04 × 10⁻² mol dm⁻³, I = 0.1 mol dm⁻³ (KNO₃) at 50 °C. The reaction takes place via an outer sphere association between Cr³⁺ and l-Dopa followed by chelation. The product was characterized by physicochemical and infrared spectroscopic methods. The antiparkinsonian activity of the product was found to be higher than that of l-Dopa.     

The influence of humic acid on the pH-dependent sorption of americium(III) onto kaolinite

This work investigates the sorption of americium [Am(III)] onto kaolinite and the influence of humic acid (HA) as a function of pH (3–11). It has been studied by batch experiments (V/m = 250:1 mL/g, C _Am(III) = 1 × 10⁻⁵ mol/L, C _HA = 50 mg/L). Results showed that the Am(III) sorption onto the kaolinite in the absence of HA was typical, showing increases with pH and a distinct adsorption edge at pH 3–5. However in the presence of HA, Am sorption to kaolinite was significantly affected. HA was shown to enhance Am sorption in the acidic pH range (pH 3–4) due to the formation of additional binding sites for Am coming from HA adsorbed onto kaolinite surface, but reduce Am sorption in the intermediate and high pH above 6 due to the formation of aqueous Am-humate complexes. The results on the ternary interaction of kaolinite–Am–HA are compared with those on the binary system of kaolinite–HA and kaolinite–Am and adsorption mechanism with pH are discussed. Effect of different molecular weight of HA, with three HA fractions separated by ultrafiltration techniques, on the Am sorption to kaolinite were also studied. The results showed that the enhancement of the sorption of Am onto kaolinite at the acidic pH conditions (pH 3–4) was higher with HA fractions of higher molecular weight. Also, the Am sorption over a pH range from 6 to 10 decreased with decreasing molecular weight of HA.     

Chiral ligand-exchange chromatography on an RP HPLC column coated with a new chiral selector derived froml-spinacine

Summary A commercial reversed-phase (RP) C₁₈ HPLC column has been dynamically coated with the chiral selectorN ^τ-n-decyl-l-spinacine and then loaded with copper(II) ions. Several racemic mixtures of underivatized amino acids and oligopeptides were resolved on the column by chiral ligand-exchange chromatography. The most important experimental conditions affecting column efficiency, retention, and selectivity (temperature and mobile phase flow rate and composition) were extensively investigated.     

A Temperature and Salt-Tolerant <Emphasis Type="SmallCaps">l</Emphasis>-Glutaminase from Gangotri Region of Uttarakhand Himalaya: Enzyme Purification and Characterization

Purification and characterization of halotolerant, thermostable alkaline l-glutaminase from a Bacillus sp. LKG-01 (MTCC 10401), isolated from Gangotri region of Uttarakhand Himalaya, is being reported in this paper. Enzyme has been purified 49-fold from cell-free extract with 25% recovery (specific activity 584.2 U/mg protein) by (NH₄)₂SO₄ precipitation followed by anion exchange chromatography and gel filtration. Enzyme has a molecular weight of 66 kDa. l-Glutaminase is most active at pH 11.0 and stable in the pH range 8.0–11.0. Temperature optimum is 70 °C and is completely stable after 3 h pre-incubation at 50 °C. Enzyme reflects more enhanced activity with 1–20% (w/v) NaCl, which is further reduced to 80% when NaCl concentration was increased up to 25%. l-Glutaminase is almost active with K⁺, Zn²⁺, and Ni²⁺ ions and K _m and V _max values of 240 μM and 277.77 ± 1.1 U/mg proteins, respectively. Higher specific activity, purification fold, better halo-tolerance, and thermostability would make this enzyme more attractive for food fermentation with respect to other soil microbe derived l-glutaminase reported so far.