Removal of heavy metals from aqueous solutions using activated carbon prepared from Cicer arietinum

Removal of Cu²⁺, Cd²⁺, Pb²⁺, and Zn²⁺ from aqueous solutions by activated carbon prepared from stems and seed hulls of Cicer arietinum, an agricultural solid waste, has been studied. The influence of various parameters, such as pH, contact time, adsorbent dose, and initial concentration of metal ions on removal was evaluated. The activated carbon was characterized by FT-IR spectroscopy, X-ray diffraction, and elemental analysis. Sorption isotherms were studied using Langmuir and Freundlich isotherm models. All experimental sorption data were fitted to the sorption models using nonlinear least-squares regression. The maximum adsorption capacity values for activated carbon prepared from Cicer arietinum waste for metal ions were 18 mg g^?1 (Cu²⁺), 18 mg g^?1 (Cd²⁺), 20 mg g^?1 (Pb²⁺), and 20 mg g^?1 (Zn²⁺), respectively. The Freundlich isotherm model fit was best, followed by the pseudo-second-order kinetic model. Desorption studies were carried out with dilute hydrochloric acid for quantitative recovery of the metal ions and for regeneration of the adsorbent.     

Single and combined effects of heavy metals and hormones on lysosomes of haemolymph cells from the mussel Mytilus galloprovincialis

Effects of heavy metals on lysosomes were studied in living cells from the mussel (Mytilus galloprovincialis Lam.). Haemolymph cells were obtained from the mussel adductor muscle, stained with neutral red (NR), and analysed by digital imaging to evaluate NR retention times within lysosomes. Exposure to Hg²⁺, Cd²⁺ and Cu²⁺ induced a reduction of NR retention time, indicating lysosomal membrane destabilisation. The intensity of these effects was correlated with the metal affinity for sulfhydryls. In contrast, Zn²⁺ showed no effect on lysosomes. Moreover, 200 μM Zn²⁺ protected lysosomes against the effects of Cd²⁺ and Cu²⁺, but not against Hg²⁺. Cell loading with the fluorescent pH probe Lyso Sensor followed by digital imaging showed a rise of lysosomal pH induced by Cd²⁺ and Hg²⁺, while Zn²⁺ prevented the effect of Cd²⁺ and also partially that of Hg²⁺. The different protective effect of Zn²⁺ against Hg²⁺ suggests a dual action of Hg²⁺ on lysosomes, possibly involving both membrane destabilisation and proton pump inhibition. Cell exposure to 17 β-estradiol also caused a reduction of NR retention time, which was synergistic to that of Hg²⁺. This suggests a common pathway between metals and hormone, possibly involving Ca²⁺ signaling. Received: 17 November 1999 / Accepted: 29 June 2000     

Immobilized Lentinus edodes residue as absorbent for the enhancement of cadmium adsorption performance

To investigate the potential use of Lentinus edodes (L. edodes) residue for Cd²⁺ adsorption, poly alcohol Na alginate (PVA) was applied to immobilize it. The parameters including contact time, pH, adsorbent dosages, and coexisting metal ions were studied. The suitable pH for immobilized L. edodes was 4?C7 wider than that for raw L. edodes (pH 6?C7). In the presence of Pb²⁺ concentration varying from 0 to 30 mg·L^?1, the Cd²⁺ adsorption ratios declined by 6.71% and 47.45% for immobilized and raw L. edodes, respectively. While, with the coexisting ion Cu²⁺ concentration varied from 0 to 30 mg·L^?1, the Cd²⁺ adsorption ratios declined by 12.97% and 50.56% for immobilized and raw L. edodes, respectively. The Cd²⁺ adsorption isotherms in single-metal and dual-metal solutions were analyzed by using Langmuir, Freundlich, and Dubinin-Radushkevich models. The Cd²⁺ adsorption capacities (q _m) in single-metal solution were 6.448 mg·L^?1 and 2.832 mg·L^?1 for immobilized and raw L. edodes, respectively. The q _m of immobilized L. edodes were 1.850 mg Cd·g^?1 in Cd²⁺ + Pb²⁺ solution and 3.961 mg Cd·g^?1 in Cd²⁺ + Cu²⁺ solution, respectively. The Cd²⁺ adsorption processes subjected to both adsorbents follow pseudo-second-order model. Mechanism study showed the functional group of L. edodes was -OH, -NH, -CO, and PVA played an important role in metal adsorbing. Mining wastewater treatment test showed that PVA-SA-immobilized L. edodes was effective in mixed pollutant treatment even for wastewater containing metal ions in very low concentration.     

Biosorption of zinc ions from aqueous solution by the microalga Scenedesmus obliquus

Aquatic environments are often exposed to toxic heavy metals, which gain access to the food chain via microalgae and may cause severe problems at higher trophic levels. However, such a metabolic specificity can be taken advantage of in bioremediation strategies. The potential of a novel wild strain of Scenedesmus obliquus, previously isolated from a heavy metal-contaminated site in northern Portugal, to remove Zn from aqueous solutions was thus studied, using several initial concentrations. The removal extent reached its maximum by 1 day: 836.5 mg Zn/g biomass, at the initial concentration of 75 mg/L, mainly by adsorption onto the cell surface. Comparative studies encompassing a commercially available strain of the same microalgal species led to a maximum removal extent of only 429.6 mg Zn/g biomass, under identical conditions. Heat-inactivated cells permitted a maximum removal of 209.6 mg Zn/g biomass, at an initial concentration of 50 mg Zn/L. The maximum adsorption capacity of Zn, estimated via Langmuir’s isotherm, was 330 mg Zn/g biomass. Finally, Zn removal was highest at pH 6.0–7.0. It was proven, for the first time, that such a wild microalga can uptake and adsorb Zn very efficiently, which unfolds a particularly good potential for bioremediation. Its performance is far better than similar (reference) species, especially near neutrality, and even following heat-treatment.     

Application of Zr(IV) tungstate for removal of metal ions from aqueous solutions

A heteropolyacid Zr(IV) tungstate-based cation exchanger has been synthesized. An amorphous sample, prepared at pH 1.2 and having a Na⁺ ion exchange capacity of 0.92?meq?g^?1, was selected for further studies. Its physicochemical properties were determined using Fourier transform infrared spectrometer, X-ray diffraction, thermogravimetric, and scanning electron studies. To understand the cation exchange behavior of the material, distribution coefficients (K _d) for metal ions in various solvent systems were determined. Some important binary separations of metal ions, namely Mg²⁺–Bi³⁺, Cd²⁺–Bi³⁺, Fe³⁺–Bi³⁺, Th⁴⁺–Bi³⁺, and Fe³⁺–Zn²⁺, were achieved on such columns. The practical utility of these separations was demonstrated by separating Fe³⁺ and Zn²⁺ ions quantitatively in commercial pharmaceutical formulation. The cation exchanger has been successfully applied also for the treatment of industrial wastewater and a synthetic mixture. All the results suggests that Zr(IV) tungstate has excellent potential for the removal of metals from aqueous systems using packed columns of this material.     

Isolation and characterisation of cadmium-resistant bacteria from an industrially polluted coastal ecosystem on the southeast coast of India

A total of 35 bacterial strains were isolated from the industrially polluted Cuddalore coast, on the southeast coast of India. Of these, 17 strains were cadmium resistant and the remainder were sensitive. Six strains (C-1, C-8, C-10, C-12, C-14 and N-1) were selected based on high levels of cadmium tolerance (>150 mg L^?1) and were termed highly cadmium-resistant bacteria (HCRB). These HCRB were identified on the basis of morphological, biochemical and partial sequencing of their 16S rRNA genes. The antibiotic-susceptibility patterns and minimum inhibitory concentrations (MIC) of different metals (Cu²⁺, Pb²⁺ and Zn²⁺) against each HCRB were determined. Among the isolates, C-14 showed high degrees of metal and antibiotic resistance compared with other HCRB. Growth rates of HCRB at two different Cd²⁺ concentrations (50 and 100 mg L^?1) and under different metal conditions (Cd²⁺, Cu²⁺ and Pb²⁺) were also investigated. HCRB growth rates were lower in the metal-treated condition than in the untreated condition. Isolates C-14 and N-1 removed>80% of Cd²⁺ from cadmium-treated broth. However, isolate C-14 removed 92.3% of Cd²⁺ compared with 86.5% for isolate N-1. Bacteria showing residual growth rates under metal stress conditions might be useful in metal removal applications under growing conditions.     

Synthesis and characterization of an ion exchanger based on tamarind kernel powder and its application for removal of some metal ions from industrial effluents

A resin synthesized from tamarind kernel powder possesses high selectivity for metal ions. Distribution coefficients for some metal ions has been determined by the batch method. The influence of pH on ion exchange capacity and K _d value of metal ions were studied. The resin has been characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis, chemical composition and ion exchange capacity (IEC). The selectivity order is Pb²⁺?>?Cu²⁺?>?Fe²⁺?>?Zn²⁺?>?Ni²⁺. Removal of metal ions from the aqueous solution and from effluents of a steel mill has been studied.     

Polarographic investigations on the complexation of cadmium and zinc by thiol peptides

Complex formation of Cd²⁺ and Zn²⁺ with thiol derivatives has been investigated by differential pulse polarography. The binding of Cd²⁺ and Zn²⁺ with cysteine (CySH), glutathione (GSH) and the model peptide N‐acetyl‐cysteine‐methylamide (ASH) reveals different stoichiometry. Thus, Cd²⁺ forms 1:1 and 1:2 complexes with CySH while 1:2 and 1:4 complexes have been observed with GSH and ASH, respectively. Overall formation constants of Cd²⁺ with CySH (Iogβ ₂ 15.3) and with GSH (Iogβ5₂ 14.4) have been estimated using competitive complexation with nitrilotriacetic acid (NTA). Investigation of competition between Zn²⁺ and Cd²⁺ for the thiol complexation has underlined the role played by the amino group in CySH for the stabilization of Zn complexes in contrary to Cd complexes.     

In vitro toxic effects of heavy metals on fungal growth and phosphate-solubilising abilities of isolates obtained from Phragmites australis rhizosphere

In this research, we evaluated the toxic effect of metal ions on mycelial growth and phosphate-solubilising activity of soil-borne micromycetes isolated from the Phragmites australis rhizosphere using Pikovskaya-agar plates supplemented with four metal concentrations. The diameter growth rate (DGR) decreased as the metal concentration rise for all tested fungi. Trichoderma atroviride had the fastest growth rate (1.48?cm²?day^?1) and was the least susceptible to Al³⁺, Cd²⁺, Cr³⁺, Cu²⁺ and Pb²⁺ with a median effective concentration (MEC₅₀) of 12.19, 0.48, 4.51, 11.44 and 50.05?mM, respectively. Aspergillus japonicus was the most tolerant to Co²⁺, Ni²⁺ and Zn²⁺, with MEC₅₀ values of 3.36, 1.095 and 2.34?mM, respectively. Penicillium italicum was the most tolerant to Cr6⁺ (MEC₅₀?=?0.677?mM). The ability to solubilise phosphate remained, despite the decrease in the DGR, and P. italicum and Penicillium dipodomyicola had the highest Phosphate Solubilisation Indexes (PSIs) at 1.97 and 2.12, respectively. In particular, P. italicum recorded the highest PSI of all the studied isolates at 0.62?mM Cr³⁺ (PSI?=?4.74). A. japonicus and T. atroviride were the most tolerant isolates to all tested metals, which suggests that these isolates are promising candidates for further study with regard to mycoremediation and biofertilisation of metal-polluted soils.     

Immobilization of some metals in contaminated sludge by zeolite prepared from local materials

In this study, zeolite was prepared from cheap local Egyptian clay (kaolin) and characterized by X-ray diffraction, X-ray fluorescence spectroscopy, SEM. The prepared zeolite was used as a binder for immobilization of the metals Cd²⁺, Cu²⁺, Ni²⁺, Pb²⁺, and Zn²⁺ in contaminated sewage sludge. Different leaching tests were conducted to determine the efficiency of the prepared zeolite for metal stabilization. The leaching of the metals from stabilized sludge decreased as the zeolite amount increased. It was found that 10% of zeolite is sufficient for the stabilization of all metal ions under investigation. It was suggested that the metal uptake mechanism by zeolite was by an ion-exchange mechanism. Examination of the solidified sample for its compressive strength after curing for 28 days yielded a value of 0.83?MPa, which indicates that the treated sludge was well solidified and safe to be used in a wide variety of applications, for instance as a raw material for pavement blocks.     

Removal of metal ions from aqueous solution by chelating polymeric hydrogel

Polysaccharide natural seed coat from the tree Magonia pubescens, in the form of hydrogel was used to remove metals in aqueous solution. Swelling tests indicate that seed coat presents hydrogel behavior, with maximum water absorption of 292 g water/g. Adsorption experiments performed using Na⁺, Mg²⁺, K⁺, Ca²⁺, Cr³⁺, Fe³⁺ and Zn²⁺ demonstrated that the polysaccharide structure has a high capacity to extract these ions from the aqueous solution. Scanning electron microscopy revealed significant morphological changes of the material before and after water contact. Differential scanning calorimetry measurements indicate a signal shift of the water evaporation temperature in the material with adsorbed zinc. X-ray photoelectron spectroscopy analysis combined with theoretical studies by the density functional theory and on Hartree–Fock (HF) level evidence that the metallic ions were adsorbed through coordination with hydroxyl groups of polysaccharide. In the case of Zn²⁺ the lowest HF energy was observed for the tetracoordination mode, where Zn²⁺ is coordinated by two hydroxyl groups and two water molecules.     

Cd2+ adsorption on alkaline-pretreated diatomaceous earth: equilibrium and thermodynamic studies

Naturally occurring diatomaceous earth was modified by alkaline pretreatment, and its effectiveness for Cd²⁺ removal from contaminated water was investigated. Batch experiments were carried out to determine Cd²⁺ adsorption capacity and the efficiency of the sorption process under different experimental conditions. Experimental data showed good fitting to Langmuir and Freundlich isotherms models. The Cd²⁺ maximum adsorption capacity was 0.058 mmol g⁻¹ for raw diatomite and increased to 0.195 mmol g⁻¹ for alkaline-pretreated diatomite with efficiency higher than 96% (diatomite dose 2.5 g L⁻¹, pH 6). Adsorption of Cd²⁺ to alkaline-pretreated diatomite increased as the temperature increased. Thermodynamic parameters were calculated to evaluate the feasibility of the adsorption process at different temperatures. The adsorption process was spontaneous and endothermic. The interaction between Cd²⁺ ions and diatomite surface was weak enough to be considered as physical sorption, confirmed by the low value of activation energy.     

重金属对禾谷镰刀菌的生长及毒素合成的影响

以禾谷镰刀菌为研究对象,研究了常见重金属(Cu、Pb、Zn、Cd)对禾谷镰刀菌菌株生长及其毒素合成的影响。研究结果表明,重金属对菌株的生长和其产毒能力均产生影响。Cu~(2+)和Cd~(2+)对菌株生长影响较大,随着浓度的增加对生长的抑制作用增强,当离子浓度分别为20 mg·L~(-1)和40 mg·L~(-1)时,能够完全抑制菌株生长。Zn~(2+)在0~160 mg·L~(-1)浓度范围内促进菌株生长,在10 mg·L~(-1)浓度下促进毒素合成。Pb~(2+)在察氏培养基中对菌株生长的影响没有明显规律可循,但是,随着Pb~(2+)浓度增加,抑制毒素合成作用增强。     

Inhibition of Na+/K+-ATPase and of active ion-transport functions in the gills of the shore crab Carcinus maenas induced by cadmium

Inhibition of Na⁺/K⁺-ATPase from gill plasma membranes of the shore crab Carcinus maenas by cadmium was investigated and compared with inhibitory effects by known antagonists (ouabain and Ca²⁺). For comparative considerations the Cd²⁺-inhibition of the enzyme from dog kidney was also tested. Na⁺/K⁺-ATPase from dog kidney and from crab gill differed greatly in sensitivity against ouabain. The inhibition constant K _i of the dog enzyme amounted to 9.1 × 10⁻⁷ mol l⁻¹, i.e. more than 300-fold smaller than the K _i of 2.9 × 10⁻⁴ mol l⁻¹ determined for the crab enzyme. Ca²⁺ inhibited the activity of Na⁺/K⁺-ATPase from crab gill plasma membranes with a K _i of 4.3 × 10⁻⁴ mol l⁻¹. The Na⁺/K⁺-ATPase from crab gill was inhibited by Cd²⁺ with a K _i of 9.1 × 10⁻⁵ mol l⁻¹. Cd²⁺ inhibited the Na⁺/K⁺-ATPase from dog kidney with a K _i (6.4 × 10⁻⁵ mol l⁻¹) comparable to that observed in the crab gill enzyme. Under experimental conditions Cd²⁺-inhibition of Na⁺/K⁺-ATPase was irreversible. Repeated washing, centrifugation and homogenization of the plasma membranes (four times) with Cd²⁺-free buffer did not restore any activity lost in the presence of 1 × 10⁻³ mol l⁻¹ Cd²⁺. Since ouabain-insensitive (nonspecific) ATPases in the plasma membrane fraction of crab gills were inhibited by Cd²⁺ in the same way as Na⁺/K⁺-ATPase, the heavy metal is considered as an unspecific ATPase inhibitor. Comparing these results with literature data on Cd²⁺-binding to electrophoretically separated proteins suggests that Na⁺/K⁺-ATPase is a Cd²⁺-binding enzyme. The results obtained on Na⁺/K⁺-ATPase were reflected by Cd²⁺-inhibition of the branchial ion-transport functions depending on this enzyme. The transepithelial short-circuit current of isolated gill half lamellae, a direct measure of area-specific active ion uptake, and the transepithelial potential difference of isolated, perfused whole gills, also indicative of active ion uptake, were inhibited by the heavy metal in a time- and dose-dependent mode. Remarkably these inhibitions were also irreversible. These findings are ecologically and biomedically significant: even when the actual environmental or tissue concentrations measured are low, biological microstructures such as Na⁺/K⁺-ATPase may accumulate the heavy metal by tight binding over prolonged periods until the first inhibitory effects occur. Received: 25 June 1997 / Accepted: 25 August 1997     

Sorption of cadmium(II) and copper(II) by soil humic acids: temperature effects and sorption heterogeneity

Sorption by humic acids is known to modify the bioavailability and toxicity of metals in soils and aquatic systems. The sorption of cadmium(II) and copper(II) to two soil humic acids was measured at pH 6.0 using ion-selective electrode potentiometric titration at different temperatures. Sorption reactions were studied with all components in aqueous solution, or with the humates in suspension. Adsorption reactions were described using a multiple site-binding model, and a model assuming a continuous log-normal distribution of adsorption constants. Adsorption of Cu²⁺ was more favourable than adsorption of Cd²⁺. The log-normal distribution model provided the closest fit to observations and allowed parameterisation of adsorption data using a mean adsorption constant (log K _μ). Sorption of Cd²⁺ to dissolved humic acids increased slightly in extent and sorption affinity with increasing temperature, but the effect was small (log K _μ 2.96–3.15). A slightly greater temperature effect occurred for sorption of Cd²⁺ to solid-phase humic acids (log K _μ 1.30–2.08). Sorption of copper(II) to both aqueous- and colloidal-phase humates showed more pronounced temperature dependence, with extent of sorption, and sorption affinity, increasing with increasing temperature (log K _μ 3.4–4.9 in solution and 1.4–4.5 in suspension). The weaker adsorption of Cd²⁺ than Cu²⁺, and smaller temperature effects for dissolved humates than suspended humates, suggested that the observed temperature effects had a kinetic, rather than thermodynamic, origin. For any metal-to-ligand ratio, free metal ion concentration, and by inference metal bioavailability, decreased with increasing temperature. The consistency of the data with kinetic rather than thermodynamic control of metal bioavailability suggests that equilibrium modelling approaches to estimating bioavailability may be insufficient.     

Preconcentration of some trace metal ions on coated alumina modified by 1-((6-(-(2-hydroxynaphthalen-1-yl)methyleneamino) hexylimino) methyl) naphthalen-2-ol

A sensitive and efficient method for preconcentration of trace amounts of some metal ions such as Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺, Pb²⁺, Cd²⁺, Cr³⁺, and Fe³⁺ ions based on modification of sodium-dodecyl-sulphate (SDS) coated alumina with 1-(6-(-(2-hydroxynaphthalen-1-yl) methyleneamino) hexylimino) methyl) naphthalen-2-ol (HNMAHN) is reported. The method is based on the uptake of these ions following their chelation with HNMAHN and their recovery using a suitable eluent. The influence of parameters such as pH, concentration of ligand and amount of coated alumina, SDS concentration, eluent (type and concentrations), and elution volume on metal ion recoveries are investigated. The preconcentration factor is 150 (10?mL elution volume) for a 1500?mL sample volume. The method has been successfully applied for extraction and determination of these ions content in some real samples. Extraction efficiency is generally >95% with low relative standard deviations between 1.8% and 2.4 %.     

Molecular dynamics of stability and structures in phytochelatin complexes with Zn,Cu, Fe,Mg, and Ca: Implications for metal detoxification

Phytochelatins, or (γ-glutamyl-cysteine) _n -glycine, are specialized peptides produced by plants and algae to mitigate toxic metal exposure, for instance in response to high levels of metals such as Cu, Cd, and Zn. Stability constants and structural characterization of metal–phytochelatin complexes are lacking. This information is required to gain mechanistic insights on the metal selectivity of phytochelatins. Here, we studied structural coordination and thermodynamic stability by performing molecular dynamics simulations of a fully hydrated phytochelatin molecule complexed with Ca²⁺, Mg²⁺, Fe²⁺, Zn²⁺, and Cu²⁺. Our results predict the following decreasing order for the thermodynamic stability of the phytochelatin complexes: Zn²⁺ ≥ Cu²⁺ ≥ Fe²⁺ > Mg²⁺ > Ca²⁺. The favorable binding energies with Zn²⁺ and Cu²⁺ over the other metal cations can be explained by shorter binding distances and greater coordination from carboxylate and keto O atoms. Conformational rearrangement of phytochelatin following metal chelation was captured by monitoring changes in the solvent-accessible volume. Accessibility of solvent molecules to the phytochelatin structure was inversely proportional to the distance between the coordinated ligands and the chelated metal. These new findings demonstrate the influence of the metal–phytochelatin structure on the metal-binding thermodynamics and the phytochelatin conformation, both of which are important to evaluate the intracellular role of phytochelatin in mediating algal response to toxic heavy metal exposure.     

镉和铅对菲律宾蛤仔脂质过氧化及抗氧化酶活性的影响

为了研究亚致死浓度的重金属对海洋贝类的毒性效应,探讨其可能的作用机理,在实验生态条件下以菲律宾蛤仔(Ru-ditapes philip pinarum)为目标生物,采用半静态毒性实验方法,研究了不同浓度Cd2+(0.0948、0.237和0.474mg·L-1)和Pb2+(0.276、0.690和1.380mg·L-1)对菲律宾蛤仔鳃和消化腺组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性以及丙二醛(MDA)含量的影响。结果显示:(1)鳃和消化腺中的抗氧化酶及MDA的变化呈现出类似的趋势,在胁迫初期,各浓度处理组菲律宾蛤仔鳃和消化腺组织SOD和CAT与对照组相比活性都显著升高(P<0.01),呈现出明显的诱导效应,而MDA含量与对照组相比差异不显著(P>0.05);随着胁迫时间的延长,高浓度Cd2+(0.474mg·L-1)和Pb2+(1.380mg·L-1)处理组中SOD、CAT活性快速下降,与对照组相比差异显著;低浓度处理组中的抗氧化活性虽然也较对照组有所下降,但总体下降幅度不如高浓度组明显,并且所历暴露时间较长。各浓度处理组中MDA含量变化一致,均呈现出先升高后降低的趋势,且含量均高于对照组。(2)通过相关性分析,菲律宾蛤仔消化腺组织中的SOD活性与Cd2+浓度的相关性大于鳃组织,与Pb2+浓度的相关性则小于鳃组织;消化腺组织中的CAT活性与Cd2+、Pb2+浓度呈抛物线型相关,与鳃组织CAT活性相关性不十分显著。这说明消化腺组织中SOD活性对Cd2+的敏感性大于鳃组织,消化腺组织中CAT活性对Cd2+、Pb2+的敏感性大于鳃组织。因此,菲律宾蛤仔消化腺中SOD、CAT对水环境中的重金属反应敏感,且存在一定的剂量-效应关系,消化腺组织中SOD和CAT活性与其他敏感性指标一起可以作为指示早期海洋重金属污染的生物标志物。     

Purification and some properties of β-N-acetyl-D-glucosaminidase from prawn (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

The -N-acetyl-D-glucosaminidase (NAGase, EC 3.2.1.52) from prawn (Penaeus vannamei) was purified by extraction with 30% ethanol solution and ammonium sulfate fractionation, then chromatographed on Sephadex G-100 followed by DEAE-cellulose (DE-32) columns. The purified enzyme determined to be homogeneous by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The specific activity of the purified enzyme was 1,560 U mg^–1. Enzyme molecular weight was determined to be 105,000 Da; it contained two subunits of the same mass (45,000 Da). The pI value was calculated to be 4.8 by isoelectric focusing. The optimum pH and optimum temperature of the enzyme for the hydrolysis of pNP--D-GlcNAc (enzyme substrate) were determined to be pH 5.2 and 45°C, respectively. The behavior of the enzyme during hydrolysis of pNP--D-GlcNAc followed Michaelis–Menten kinetics, with K_m=0.254 mM and V_m=9.438 M min^–1, at pH 5.2 and 37°C. The stability of the enzyme was investigated, and the results showed that the enzyme was stable in a pH range from 4.2 to 10.0 and at temperatures <40°C. The effects of metal ions on the enzyme were also studied. Li⁺, Na⁺ and K⁺ had no influence on enzyme activity. Mg²⁺, Ca²⁺ and Mn²⁺ activated the enzyme, while Ba²⁺, Zn²⁺, Co²⁺, Cd²⁺, Hg²⁺, Pb²⁺ Cu²⁺, Fe³⁺ and Al³⁺ showed various degrees of inhibitory effects on the enzyme.Communicated by O. Kinne, Oldendorf/Luhe     

Bioleaching of cadmium and nickel from synthetic sediments by Acidithiobacillus ferrooxidans

The microbial leaching process was evaluated for the treatment of synthetic sediments contaminated with cadmium and nickel sulfides. A series of batch leaching experiments was conducted to compare metal solubilization in sediment inoculated with Acidithiobacillus ferrooxidans -inoculated sediments to that in sterile control sediment. The rate and extent of metal solubilization were significantly higher in A. ferrooxidans -inoculated reactors than in acidified sterile reactors. The efficiency of cadmium (Cd) solubilization (80) in the bioleaching process was higher than that of nickel (Ni) solubilization (60). The performance of leaching reactors containing only culture supernatants was comparable to that of A. ferrooxidans -inoculated reactors, indicating that indirect non-contact leaching by the products of microbial metabolism is the predominant mechanism for metal solubilization rather than direct microbial sulfide oxidation. Moreover, the similar (60–75%) extents of Cd²⁺ leaching with A. ferrooxidans , cell-free filtrate, and Fe³⁺ suggest that abiotic oxidation of CdS by Fe³⁺ controls the overall leaching rate, and the role of A.␣ferrooxidans is most likely not to oxidize CdS mineral directly but to regenerate Fe³⁺ as an oxidant.