Self-immobilization of Recombinant Caulobacter crescentus and Its Application in Removal of Cadmium from Water

Microbial biofilms can be valuable in many biotechnological applications, including bioremediation. We have previously constructed a recombinant strain of Caulobacter crescentus JS4022/p723-6H by inserting a hexahistidine peptide to a permissive site of the host surface layer (S-layer) protein RsaA. This engineered strain is highly effective in removal of cadmium from water as free cells. In this communication, we examined the biofilms formed by self-immobilized JS4022/p723-6H and evaluated their ability to retrieve cadmium from contaminated water samples. According to light and electron microscopic observations, JS4022/p723-6H cells developed a uniform monolayer biofilm on borosilicate surfaces through their intrinsic appendage, a stalk with an adhesive holdfast. The density of the biofilms reached a maximum after 48 h of incubation and was not affected by exposure to at least 1 ppm cadmium. When 0.4 ppm Cd(II) was added to the growth medium, immobilized JS4022/p723-6H removed 76.9% of the total metal, whereas the control strain only removed 13.5%. When a water sample collected from Lake Erie was spiked with various amounts of CdCl₂, immobilized JS4022/p723-6H was able to sequester 44～51% of the total metal, compared to 37～42% accumulated by the control strain. By combining two powerful techniques, cell surface display and self-immobilization, we achieved complete separation of dissolved heavy metals from contaminated water in a single step. This study laid down the foundation to cost-effectively construct large-scale bioreactors with high efficiency and specificity to retrieve environmental contaminants from water.     

Complexation Studies of N,N′-ethylenedi-L-cysteine with Some Metal Ions

As part of a search for environmentally friendly metal chelating ligands, the stability constants of N, N′-ethylenedi-L-cysteine (EC) complexes with Ca(II), Cu(II), Mg(II) and Mn(II) were determined by potentiometry with a glass electrode in aqueous solutions containing 0.1 mol⋅L⁻¹ KCl at 25 °C. Final models are proposed. For the Ca(II)–EC system, the overall stability constants are log ₁₀ β _CaHL=14.53±0.03, log ₁₀ β _CaL=4.79±0.01 and log ₁₀ β _CaL2=8.38±0.04. For the M(II)–EC systems, where M=Cu(II) or Mg(II), the overall stability constants are log₁₀ β _CuHL=31.19±0.02 and log ₁₀ β _CuL=27.02±0.06 for Cu(II), and are log ₁₀ β _MgHL=14.84±0.02 and log ₁₀ β _MgL=6.164±0.008 for Mg(II). For the Mn(II)–EC system, the overall stability constant is log ₁₀ β _MnL=10.12±0.01. Metal–chelate speciations simulations showed that EC is an efficient chelating agent for Cd(II), Co(II), Cu(II), Ni(II), Pb(II) and Zn(II) for pH≥7.     

X-Ray Diffraction and Raman Spectroscopic Study of Bis-(Pyridine Base) Complexes of Cadmium(II) Halogenides

Summary.  The molecular structures of bis-(pyridine base) complexes of cadmium(II) chloride and bromide, where the pyridine base is pyridine ( py), 3-methylpyridine (3-Me-py), 4-methylpyridine (4-Me-py), and 4-ethylpyridine (4-Et-py), were investigated by means of single-crystal X-ray diffraction and Raman spectroscopy. The crystal structures of CdCl₂ py ₂ (1), CdCl₂(3-Me-py)₂ (2), and CdCl₂(4-Me-py)₂ (3) were determined. All crystals are monoclinic; 1: a = 17.784(2), b = 8.666(1), c = 3.8252(7) ?, β = 91.54(1)°, space group: P2₁/n; 2: a = 11.89(1), b = 14.41(1), c = 3.874(6) ?, β = 92.3(1)°, space group: P2₁/a; 3: a = 21.091(2), b = 3.8884(5), c = 18.2317(3) ?, β = 113.64(1)°, space group: C2/c. The structures were refined to R/R _w values (%) of 3.2/5.5, 3.0/5.0, and 3.4/5.1 for 1–3. All cadmium atoms are octahedrally coordinated with the chloride ions forming infinite di-μ-chloro polymeric linear chains and the nitrogen atoms of the pyridine base in trans configuration. The Cd chains are oriented along the c-axis in 1 and 2 and along the b-axis in 3. The crystal structures indicate the absence of a peculiar interaction between the polymeric chains. The Raman spectra of eight complexes were measured in the range of 550–50 cm⁻¹, and the Raman peaks originating from cadmium-halogen vibrations were assigned. The Raman spectra of 1 and 2 are quite alike in the lattice mode vibration region. The resemblance of the cadmium-halogen vibration peaks indicates the same halogenide ion bridged octahedral structure for all complexes. Received March 27, 2001. Accepted (revised) June 19, 2001     

Electrochemical solid phase micro-extraction and determination of salicylic acid from blood samples by cyclic voltammetry and differential pulse voltammetry

The electrochemical solid phase micro-extraction of salicylic acid (SA) at graphite-epoxy-composed solid electrode surface was studied by cyclic voltammetry. SA was oxidized electrochemically in pH 12.0 aqueous solution at 0.70 V (vs. saturated calomel electrode) for 7 s. The oxidized product shows two surface-controlled reversible redox couples with two proton transferred in the pH range of 1.0∼6.0 and one proton transferred in the pH range of 10.0∼13.0 and is extracted on the electrode surface with a kinetic Boltzman function of i _p = 3.473–4.499/[1 + e^{(t − 7.332)/6.123}] (χ ² = 0.00285 μA). The anodic peak current of the extracted specie in differential pulse voltammograms is proportional to the concentration of SA with regression equation of i _p = −5.913 + 0.4843 c (R = 0.995, SD = 1.6 μA) in the range of 5.00∼200 μM. The detection limit is 5.00 μM with RSD of 1.59% at 60 μM. The method is sensitive and convenient and was applied to the detection of SA in mouse blood samples with satisfactory results.     

Preparation and characterization of AuPt alloy nanoparticle–multi-walled carbon nanotube–ionic liquid composite film for electrocatalytic oxidation of cysteine

Gold–platinum (AuPt) alloy particles were fabricated directly on multi-walled carbon nanotubes (MWNT)–ionic liquid (i.e., trihexyltetradecylphosphonium bis(trifluoromethylsulfonyl)imide, [P_6,6,6,14][NTf₂]) composite coated glassy carbon electrode (GCE) by electrodeposition method. Scanning electron microscope image showed that they were well-dispersed nanocluster consisting of smaller nanoparticles, and their size was about 70 nm. X-ray diffraction experiment showed that they were single-phase alloy nanomaterial, and the calculated composition was consisting with that obtained by energy dispersive X-ray spectroscopy. The resulting modified electrode (i.e., AuPt–MWNT–[P_6,6,6,14][NTf₂]/GCE) presented high catalytic activity for the electrochemical oxidation of cysteine. The peak potential of cysteine shifted to 0.42 V (versus saturated calomel electrode) in 0.1 M H₂SO₄ and the peak current increased greatly in comparison with that on the corresponding Pt (or Au)–MWNT–[P_6,6,6,14][NTf₂]/GCE. Under the optimized conditions, the oxidation current of cysteine at 0.45 V was linear to its concentration in the range of 5.0 × 10⁻⁷ ∼ 4.0 × 10⁻⁵ M with a sensitivity of 43.8 mA M⁻¹.     

Impedance study of adsorption of iodide ions at Cd(0001) and Bi(111) electrode from various solutions with constant ionic strength

Adsorption of iodide ions at the Bi(111) and Cd(0001) electrodes from the aqueous solutions with constant ionic strength 0.1x M KI + 0.1(1−x) M KF and 0.1x M KI + 0.033(1−x) M K₂SO₄ has been studied by impedance spectroscopy. It was found that, to a first approximation, the classical Frumkin–Melik–Gaikazyan equivalent circuit with the slow diffusion-like and adsorption steps can be applied for fitting the experimental impedance data for iodide ions adsorption on Bi(111) and Cd(0001) from aqueous solutions with constant ionic strength. The modified Grafov–Damaskin circuit can be used in the region of electrode potentials, where parallel faradic processes (electroreduction of protons, oxygen traces) are probable. The more complicated Ershler equivalent circuit, taking into account the slow diffusion-like, adsorption and charge transfer steps, is not applicable for characterization of the adsorption process of I⁻ at Bi(111) and Cd(0001) electrodes.     

Formation of cuprous oxide layers in Cu(II) solutions containing gluconic acid

In accordance with thermodynamic analysis, cuprous oxide layers are formed spontaneously in the Cu|Cu(II), gluconic acid system at pH > 3.7 under open-circuit conditions. A current peak of Cu₂O reduction is observed on cathodic voltammograms at ca −0.7 V, its height being dependent on the exposure time. The analysis of the charge transferred in this region yields the rate of Cu₂O formation equal to 1.25 × 10⁻¹⁰ mol cm⁻² s⁻¹. The light perturbation of Cu electrode under open-circuit conditions results in the generation of a negative photopotential, which is indicative of n-type conductivity. The threshold wavelength is equal to ∼590 nm and is consistent with a band gap of ∼2.1 eV. Anodic photocurrents, which are observed near the open-circuit potential, decrease with cathodic polarization and change their sign at ∼0.05 V. Analysis of impedance data was performed, invoking the equivalent circuit that accounts for the two-step charge transfer. In the presence of Cu₂O, some retardation of Cu(II) reduction was found to occur with a slight increase in the admittance of the double layer. The suggestion has been made that oxide layers formed in Cu(II) gluconate solutions cannot be compact and uniformly distributed over the entire electrode surface. Relevant investigations of surface morphology support this conclusion.     

An “off–on” fluorescence probe for chromium(III) ion determination in aqueous solution

An “off–on” rhodamine-based fluorescence probe for the selective signaling of Cr(III) has been designed by exploiting the guest-induced structure transform mechanism. This system shows a sharp Cr(III)-selective fluorescence enhancement response in 100% aqueous system under physiological pH value and possesses high selectivity against the background of environmentally and biologically relevant metal ions including Cr(VI), Al(III), Fe(III), Cd(II), Co(II), Cu(II), Ni(II), Zn(II), Mg(II), Ba(II), Pb(II), Na(I), and K(I). Under optimum conditions, the fluorescence intensity enhancement of this system is linearly proportional to Cr(III) concentration from 5.0 × 10⁻⁸ to 7.0 × 10⁻⁶ mol L⁻¹ with a detection limit of 1.6 × 10⁻⁸ mol L⁻¹.     

Study of the optimal condition for electroplating of Bi2S3 thin films and their photoelectrochemical characteristics

Bismuth sulfide (Bi₂S₃) thin films were electrodeposited from non-aqueous dimethyl sulfoxide medium containing Bi(NO₃)₃ and thiourea as the precursor salts, triethanol amine as the complexing agent, and TritonX-100 as the surface active agent. The prepared films were subjected to rigorous experimentation in order to validate their potential candidature for solar cells. The films exhibited band gap energy of ∼1.3 eV and resistivity of the order of 2 × 10⁶ Ω cm at room temperature as was obtained from UV–Vis spectroscopy and four-probe measurements, respectively. Scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and energy dispersive analysis of X-ray were employed to reveal the morphology, structure, and chemical composition of the film matrix. The Bi₂S₃ films were found to be non-decomposable up to the temperature of 1,000 °C with the help of thermogravimetry–differential thermal analysis. The Nyquist and Mott–Schottky plots derived from electrochemical impedance spectroscopy measurements provided important information regarding electrical and semiconducting properties of the films. The n-type film with a donor density of the order of ∼10²³ m⁻³ displayed reasonable photoactivity under illumination and is recommended as a promising candidate for potential photoelectrochemical applications.     

Self-assembled sulfonated β-Cyclodextrin layer on gold electrode for the selective electroanalysis of dopamine

Gold electrode with self-assembled D,L-cysteine grafted β-cyclodextrin sulfonic acid (Cys-β-CD∼SO₃) layer was fabricated and used to investigate the electrochemical behavior of dopamine. The experimental results indicated that the self-assembled Cys-β-CD∼SO₃ layer modified gold electrode has selective electrochemical response to dopamine with high sensitivity and excellent tolerance of ascorbic acid, which is the most common accompanying component in biological samples. Dopamine could be accurately determined in the concentration range of 1–200 μM in the presence of ascorbic acid of 5 mM. The relative standard deviation of 1.9% (n = 5) was achieved at a dopamine concentration of 5 × 10⁻⁵ M. The proposed sensor was successfully applied to the determination of dopamine in human blood serum samples.     

Interaction of Cucurbit[n = 6∼8]urils and Benzimidazole Derivatives

The structures and optical properties of host–guest complexes produced from cucurbit[n = 6–8]urils and some benzimidazole derivatives have been investigated by ¹H NMR spectroscopy, electronic absorption spectroscopy and fluorescence spectroscopy. The experimental results reveal that calculations of A∼N_Q[n]/N_guest and I_f∼N_Q[n]/N_guest for the same association complex both support a good fit to an identical binding model. In particular, the A∼N_Q[n]/N_guest, I_f∼N_Q[n]/N_guest calculations and the ¹H NMR determinations for three Q[6]–ge(1∼3) complexes and three Q[8]–ge(1∼3) complexes all support a binding model of 1:1 and 1:2 respectively.     

Voltammetric sensor for nitrite determination based on its electrocatalytic reduction at the surface of p-duroquinone modified carbon paste electrode

A p-duroquinone (tetramethyl-p-benzoquinone) modified carbon paste electrode (DMCPE) was employed to study the electrocatalytic reduction of nitrite in aqueous solutions using cyclic voltammetry (CV), double potential-step chronoamperometry, and differential pulse voltammetry (DPV). It has found that under an optimum condition (pH 1.00), the reduction of nitrite at the surface of DMCPE occurs at a potential of about 660 mV less negative than that of an unmodified carbon paste electrode (CPE). The catalytic rate constant, k′_h, based on Andrieux and Saveant theoretical model was calculated as for scan rate 10 mV s^-1. Also, the apparent diffusion coefficient, D _app, was found as 2.5 × 10^–10 and 3.61 × 10^–5 cm² s^-1 for p-duroquinone in carbon paste matrix and nitrite in aqueous buffered solution, respectively. The values for αn_α were estimated to be −0.65 and −0.19 for the reduction of nitrite at the surface of DMCPE and CPE, respectively. The electrocatalytic reduction peak currents showed a linear dependence on the nitrite concentration, and a linear analytical curve was obtained in the ranges of 5.0 × 10^–5 M to 8.0 × 10^–3 M and 6.0 × 10^–6 M to 8.0 × 10^–4 M of nitrite concentration with CV and DPV methods, respectively. The detection limits (2σ) were determined as 2.5 × 10^–5 M and 4.3 × 10^–6 M by CV and DPV methods. This method was also applied as a simple, selective and precise method for determination of nitrite in real samples (the weak liquor from the wood and paper factory of Mazandaran province in Iran) by using a standard addition method.     

Separation and determination of cadmium and zinc as their thenoyltrifluoroacetone complexes with dibenzo-18-crown-6 by means of synergistic extraction and atomic absorption spectrometry

 A new method for the separation and determination of trace amounts of cadmium and zinc in water as their thenoyltrifluoroacetone (TTA) complexes with dibenzo-18-crown-6 (DB18C6) in o-dichlorobenzene has been established by means of synergistic extraction and back-extraction combined with atomic absorption spectrometry. The effect of various factors (synergism with TTA and DB18C6, shaking time, composition of the extracted species, and mutual separation etc.) on the extraction and back-extraction of cadmium and zinc has been in- vestigated. When the mixtures were extracted for 4 min at pH 4.9, only zinc was extracted quantitatively, whereas cadmium remained in the aqueous phase. After the phases were separated, cadmium was again extracted quantitatively at pH 7.5. Then, the two phases were each shaken with 0.05–0.1 mol/l HCl in order to back-extract cadmium and zinc from the organic phases; the ions were determined individually by atomic absorption spectrometry. In the process cadmium and zinc TTA chelates in o-dichlorobenzene form stable adducts with DB18C6 (Cd(TTA)₂ ⋅ nDB18C6 and Zn(TTA)₂ ⋅ nDB18C6, n=0∼2). The stability constants (βn) of the adducts determined by means of the curve fitting method were log β₁=4.62 and log β₂=6.74 for cadmium, and log β₁=3.48 and log β₂=5.18 for zinc. Received: 8 September 1995/Revised: 22 January 1996/Accepted: 24 January 1996     

Magnesium ion-conducting gel polymer electrolytes dispersed with fumed silica for rechargeable magnesium battery application

Effect of fumed silica dispersion on poly(vinylidene fluoride-co-hexafluoropropylene)-based magnesium ion-conducting gel polymer electrolyte has been studied using various physical and electrochemical techniques. The composite gel electrolytes are free-standing and flexible films with enough mechanical strength. The optimized composition with 3 wt.% filler offers a maximum ionic conductivity of ∼1.1 × 10⁻² S cm⁻¹ at ∼25 °C with good thermal and electrochemical stabilities. The Mg²⁺ ion conduction in the gel nanocomposite film is confirmed from the cyclic voltammetry, impedance spectroscopy, and transport number measurements. The space-charge layers formed between filler particles and gel electrolyte are responsible for the enhancement in ionic conductivity. The applicability of the gel nanocomposite to a rechargeable battery is examined by fabricating a prototype cell consisting of Mg [or Mg-multiwalled carbon nanotube (MWCNT) composite] and MoO₃ as negative and positive electrodes, respectively. The discharge capacity and the rechargeability of the cell have been improved when Mg metal is substituted by Mg-MWCNT composite. The discharge capacity of the optimized cell has found to be ∼175 mAh g⁻¹ of MoO₃ for an initial ten charge–discharge cycles.     

Nafion-based solid-state gas sensors: Pt / Nafion electrodes prepared by an impregnation-reduction method in sensing oxygen

The influences of the reductant concentration of NaBH₄ and the quantity of Pt loading on the active surface area and the sensitivity of the Pt/Nafion electrodes prepared by an impregnation-reduction method in detecting oxygen were investigated in this study. The Pt/Nafion electrodes with a Pt loading of 4.99 mg/cm², obtained at 0.0107 M Pt(NH₃)₄Cl₂ and 0.06 M NaBH₄, show maximum sensitivities of 0.0528 A/ppm and 0.0538 A/ppm obtained in O₂ concentration regions of 0–5000 and 5000–50 000 ppm, respectively. A sensing model was also proposed to illustrate the sensing phenomenon. Received: 21 January 1998 / Accepted: 10 March 1998     

Spectral, magnetic, thermal, antimicrobial, and eukaryotic DNA studies on acetone [N-(3-hydroxy-2-naphthoyl)]hydrazone complexes

Abstract  

Acetone [N-(3-hydroxy-2-naphthoyl)] hydrazone (H₂AHNH) has been prepared and its structure confirmed by elemental analysis and ¹H NMR spectroscopy. It has been used to produce diverse complexes with Co(II), Ni(II), Cu(II), Zn(II), Cd(II), and U(VI)O₂ ions. The complexes obtained have been investigated by thermal analysis, spectral studies (¹H NMR, IR, UV–visible, ESR), and magnetic measurements. IR spectra suggest that H₂AHNH acts as a bidentate ligand. The electronic spectra of the complexes and their magnetic moments provide information about geometries. The ESR spectra give evidence for the proposed structure and the bonding for some Cu(II) complexes. Thermal decomposition of the Ni(II) and Cu(II) complexes afforded metal oxides as final products. Kinetic data were obtained for each stage of thermal degradation of some of the complexes using the Coats–Redfern method. The formation of complexes in solution was studied pH-metrically and the order of their stability constants (log K) was found to be U(VI)O₂ > Cu(II) > Zn(II) > Ni(II) > Cd(II) > Co(II). Antimicrobial and eukaryotic DNA studies were carried out.     

Hydrogen-Related Paramagnetic Centers in Ge-Doped Sol-Gel Silica Induced by γ-Ray Irradiation

We have studied the generation mechanisms of H(II) paramagnetic centers in Ge-doped silica by investigating up to 10⁴ mol ppm sol-gel Ge-doped silica materials. We have considered materials with the same concentrations of Ge but that are produced by two different densification routes that give rise to different concentrations of Ge-related oxygen deficient centers (GeODC(II)). These centers are characterized by an optical absorption band at ∼5.2 eV (B_2β band) and two related emissions at ∼3.2 eV and ∼4.3 eV. The GeODC(II) content was estimated by absorption and emission measurements. The H(II) centers were induced by room temperature γ-ray irradiation and their concentration was determined by electron paramagnetic resonance measurements. The comparison between the two kinds of materials, obtained by different preparations, shows that the GeODC(II) concentration increases with the Ge content and is enhanced by vacuum densification. The comparison of irradiated samples shows that beyond the already known process of conversion of preexisting GeODC(II) into H(II) centers, another generation process of H(II) centers is effective that involves irradiation induced GeODC(II).     

Polymer supported 5,10,15,20-tetrakis(phenoxy acetic acid)porphyrin derivative for separation and preconcentration of d- and f-electron metals

5,10,15,20-tetrakis(phenoxy acetic acid) porphyrin (PAAP) was covalently linked to Merrifield chloromethylated resin. Characterization of PAAP and the modified polymeric matrix were performed by ¹H NMR, FTIR and elemental analysis. The sorbent was used for the separation and enrichment of the d-electron metals (Mn(II), Co(II), Ni(II), Cu(II) and Zn(II)) at pH 6–8 and of the f-electron metals U(VI) and Th(IV) at pH 4–5. The metals ions were preconcentrated with a concentration factor range of 115–215 and then determined by flame atomic absorption spectrometry or visible spectrophotometry using Arsenazo(III). The retained metals were eluted with 2.0 mol L⁻¹ HNO₃ in the case of the d-electron metals and 0.1/0.25 mol L⁻¹ HCl in the case of the f-electron metals. The procedure was validated by analyzing the NIST standard reference material 2709 (San Joaquin Soil). Correspondence: Melek Merdivan, Chemistry Department, Faculty of Arts and Sciences, Dokuz Eylul University, 35160 Buca, Izmir, Turkey     

A new amperometric H<Subscript>2</Subscript>O<Subscript>2</Subscript> biosensor based on nanocomposite films of chitosan–MWNTs,hemoglobin, and silver nanoparticles

A new H₂O₂ biosensor was fabricated on the basis of nanocomposite films of hemoglobin (Hb), silver nanoparticles (AgNPs), and multiwalled carbon nanotubes (MWNTs)–chitosan (Chit) dispersed solution immobilized on glassy carbon electrode (GCE). The immobilized Hb displayed a pair of well-defined and reversible redox peaks with a formal potential (E ^θ′) of −22.5 mV in 0.1 M pH 7.0 phosphate buffer solution. The apparent heterogeneous electron transfer rate constants (k _s) in the Chit–MWNTs film was evaluated as 2.58 s⁻¹ according to Laviron’s equation. The surface concentration (Γ*) of the electroactive Hb in the Chit–MWNTs film was estimated to be (2.48 ± 0.25) × 10⁻⁹ mol cm⁻². Meanwhile, the Chit–MWNTs/Hb/AgNPs/GCE demonstrated excellently electrocatalytical ability to H₂O₂. Its apparent Michaelis–Menten constant (K _M^app) for H₂O₂ was 0.0032 mM, showing a good affinity. Under optimal conditions, the biosensors could be used for the determination of H₂O₂ ranging from 6.25 × 10⁻⁶ to 9.30 × 10⁻⁵ mol L⁻¹ with a detection limit of 3.47 × 10⁻⁷ mol L⁻¹ (S/N = 3). Furthermore, the biosensor possessed rapid response to H₂O₂ and good stability, selectivity, and reproducibility.     

Solid-state Cu (II) ion-selective electrode based on polyaniline-conducting polymer film doped with copper carmoisine dye complex

A pencil graphite electrode coated by copper (II)–carmoisine dye complex in polyaniline (emeraldine base form) matrix (termed as PGE/PA/Cu-Car) was prepared and used as copper ion-selective electrode. The introduced electrode was found to have high selectivity toward copper ion (II) and exhibited wide working concentration range, low response time, and good shelf life. The sensor electrode showed a linear Nernstian response over the range of 5.0 × 10⁻⁶ to 1.0 × 10⁻¹ M with a slope of 29.7 ± 1 mV per decade change in concentration. A detection limit of 2.0 × 10⁻⁶ M was obtained. The optimum pH working range of the electrode was found to be 4.0–7.0.