nitrophenols and 3-nitrophenol

Chloronitrophenols (CNPs) constitute a group of environmental pollutants that are widely distributed in our surrounding environment due to human based activities. This group of chemicals is highly toxic to living beings due to its mutagenic and carcinogenic nature. Examples include 2-chloro-4-nitrophenol, 4-chloro-2-nitrophenol, 2-chloro-5-nitrophenol, 4-chloro-3-nitrophenol and 2,6-dichloro-4-nitrophenol. Several methods including advanced oxidation processes, adsorption and bacterial degradation have been used for degradation of CNPs. Among, bacterial degradation is an eco-friendly and effective way to degrade CNPs. Several bacterial metabolic pathways have been proposed for degradation of CNPs and their genes and enzymes have been identified in bacteria. These bacteria were able to degrade CNPs in broth culture and soil. Therefore, CNPs-degrading bacteria are suitable candidates for bioremediation of CNPs-contaminated sites. Few CNP-degrading bacteria exhibited chemotaxis towards CNPs to enhance their biodegradation. The present review summarizes recent progress in degradation of CNPs.

Topics: Biodegradation, Environmental; Humans; Nitrophenols

A very simple, as well as sensitive and selective, sensing protocol was developed on a pre-anodized graphite pencil electrode surface coated using poly(thionine) (APGE/PTH). The poly(thionine) coated graphite pencil was then used for simultaneous sensing of 3-nitrophenol (3-NP) and 4-nitrophenol (4-NP). The poly(thionine) coated electrode exhibited an enhanced electrocatalytic property towards nitrophenol (3-NP and 4-NP) reduction. Redox peak potential and current of both nitrophenols were found well resolved and their simultaneous analysis was studied. Under optimized experimental conditions, APGE/PTH showed a long linear concentration range from 20 to 230 nM and 15 nM to 280 nM with a calculated limit of detection (LOD) of 4.5 and 4 nM and a sensitivity of 22.45 µA/nM and 27.12 µA/nM for 3-NP and 4-NP, respectively. Real sample analysis using the prepared sensor was tested with different environmental water samples and the sensors exhibited excellent recovery results in the range from 98.16 to 103.43%. Finally, the sensor exposed an promising selectivity, stability, and reproducibility towards sensing of 3-NP and 4-NP.

Topics: Electrochemical Techniques; Electrodes; Graphite; Nitrophenols; Phenothiazines; Reproducibility of Results; Water

The present study reports the development of an electrochemical sensor based on sulfobutylether-β-cyclodextrin modified reduced graphene oxide hybrid (SBCD-rGO) for simultaneous detection of nitrophenol isomers. First, SBCD-rGO hybrid was synthesized and detailed characterized. Afterwards, a sensor was fabricated via the modification of glassy carbon electrode (GCE) with SBCD-rGO, and its electrochemical detection performances were also investigated. Then, the constructed electrochemical sensor was applied to detect nitrophenol isomers by voltammetry analysis. The results suggested that the sensitivities were 389.26, 280.88 and 217.19 μA/mM for p-nitrophenol (p-NP), m-nitrophenol (m-NP), and o-nitrophenol (o-NP), respectively, and their corresponding detection limits were all about 0.05 μM. Significantly, the combination of voltammetry analysis with the constructed sensor and data analysis by multiple linear regression realized the simultaneous detection of nitrophenol isomers.

Topics: beta-Cyclodextrins; Carbon; Electrochemical Techniques; Electrodes; Graphite; Isomerism; Limit of Detection; Nanostructures; Nitrophenols; Reproducibility of Results

Analysis of 4-cyanophenol and 3-nitrophenol was carried out using multi-walled carbon nanotubes-based solid-phase extraction (SPE) and capillary electrophoresis (CE) methods. Capillary electrophoresis was carried out with 18 kV voltage, 214 nm detection, and phosphate buffer (pH 7.0, 15 mM) as background electrolyte at 25 ± 1 °C temperature with 15.05 and 16.5 min migration times of 4-cyanophenol and 3-nitrophenol. The separation and resolution factors were 1.10 and 2.90. The optimized experimental conditions were 40 mg/L concentration, 1.0 g multi-walled carbon nanotubes (MWCNTs) per SPE cartridge, 5.0 mL/min flow rate of water, 0.1 mL flow rate of eluting solvent. The maximum recoveries were 91% and 98% at 0.1 mL/min flow rate of 4-cyanophenol and 3-nitrophenol. These methods were applied successfully for extraction and estimation of 4-cyanophenol and 3-nitrophenol in the municipal wastewater. The reported methods are reproducible, efficient, and practical for the estimation of these phenols in water.

Topics: Electrophoresis, Capillary; Nanotubes, Carbon; Nitrophenols; Phenols; Solid Phase Extraction; Water

Zinc molybdate (ZnMoO₄) was prepared by thermal decomposition of an oxalate complex under a controlled temperature of 500 °C. Analyses of the oxalate complex were carried out using Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). On the other hand, analyses of the synthesized zinc molybdate were carried out by X-ray diffraction (XRD), transmission electron microscopy (TEM), and Brunauer-Emmett-Teller technique (BET). The efficiency of the synthesized catalyst was tested with the reduction reaction of 3-nitrophenol (3-NP), and was also applied as a sorbent for methylene blue dye (MB) in aqueous solutions. The catalytic test of zinc molybdate shows a very high activity. The concentration reduction progress and adsorption of the dye were followed by an ultraviolet-visible (UV-vis) spectrophotometer.

Topics: Catalysis; Methylene Blue; Microscopy, Electron, Transmission; Molecular Structure; Molybdenum; Nitrophenols; Oxalates; X-Ray Diffraction; Zinc

Quantifying the extent of biodegradation of nitroaromatic compounds (NACs) in contaminated soils and sediments is challenging because of competing oxidative and reductive reaction pathways. We have previously shown that the stable isotope fractionation of NACs reveals the routes of degradation even if it is simultaneously caused by different bacteria. However, it is unclear whether compound-specific isotope analysis (CSIA) can be applied in situations where multiple pollutants are biodegraded by only one microorganism under multi-substrate conditions. Here we examined the C and N isotope fractionation of 2-nitrophenol (2-NP) and 3-nitrophenol (3-NP) during biodegradation by Pseudomonas putida B2 through monooxygenation and partial reductive pathways, respectively, in the presence of single substrates vs. binary substrate mixtures. Laboratory experiments showed that the reduction of 3-NP by Pseudomonas putida B2 is associated with large N and minor C isotope fractionation with C and N isotope enrichment factors, ε

Topics: Biodegradation, Environmental; Carbon Isotopes; Chemical Fractionation; Chromatography, High Pressure Liquid; Geologic Sediments; Isomerism; Kinetics; Nitrogen Isotopes; Nitrophenols; Oxidation-Reduction; Pseudomonas putida; Soil; Soil Pollutants

To investigate green synthesis of gold nanoparticles (AuNPs) by Trichosporon montevideense, and to study their reduction of nitroaromatics.. AuNPs had a characteristic absorption maximum at 535 nm. Scanning electron microscopy images revealed that the biosynthesized nanoparticles were attached on the cell surface. X-ray diffraction analysis indicated that the particles formed as face-centered cubic (111)-oriented crystals. The average size of AuNPs decreased from 53 to 12 nm with increasing biomass concentration. The catalytic reduction of 2-nitrophenol, 3-nitrophenol, 4-nitrophenol, o-nitrophenylamine and m-nitrophenylamine (0.1 mM) by NaBH4 had reaction rate constants of 0.32, 0.44, 0.09, 0.24 and 0.39 min(-1) with addition of 1.45 × 10(-2) mM AuNPs.. An eco-friendly approach for synthesis of AuNPs by T. montevideense is reported for the first time. The biogenic AuNPs could serve as efficient catalysts for hydrogenation of various nitroaromatics.

Topics: Catalysis; Gold; Green Chemistry Technology; Hydrogenation; Metal Nanoparticles; Nitrophenols; Trichosporon

Reliable, selective and sensitive approaches for trinitrophenol (TNP) detection are highly desirable with respect to national security and environmental protection. Herein, a simple and novel fluorescent strategy for highly sensitive and specific TNP assay has been successfully developed, which is based on the quenching of the fluorescent poly(thymine)-templated copper nanoclusters (DNA-CuNCs), through the synergetic effects of acid induction and electron transfer. Upon the addition of TNP, donor-acceptor complexes between the electron-deficient nitro-groups in TNP and the electron-donating DNA templates are formed, resulting in the close proximity between TNP and CuNCs. Moreover, the acidity of TNP contributes to the pH decrease of the system. These factors combine to dramatically quench the fluorescence of DNA-CuNCs, providing a "signal-off" strategy for TNP sensing. The as-proposed strategy demonstrates high sensitivity for TNP assay, and a detection limit of 0.03μM is obtained, which is lower than those reported by using organic fluorescent materials. More significantly, this approach shows outstanding selectivity over a number of TNP analogues, such as 2,4,6-trinitrotoluene (TNT), 2,4-dinitrotoluene (DNT), 2,4-dinitrophenol (DNP), 3-nitrophenol (NP), nitrobenzene (NB), phenol (BP), and toluene (BT). Compared with previous studies, this method does not need complex DNA sequence design, fluorescent dye labeling, or sophisticated organic reactions, rendering the strategy with additional advantages of simplicity and cost-effectiveness. In addition, the as-proposed strategy has been adopted for the detection of TNP in natural water samples, indicating its great potential to be applied in the fields of public safety and environmental monitoring.

Topics: Biosensing Techniques; Copper; DNA; Fluorescence; Fluorescent Dyes; Lakes; Nanostructures; Nitrophenols; Oxidation-Reduction; Spectrometry, Fluorescence; Water Pollutants, Chemical

An express method for determination of acidity constants of organic acids, based on the analysis of the integral transmittance vs. pH dependence is developed. The integral value is registered as a photocurrent of photometric device simultaneously with potentiometric titration. The proposed method allows to obtain pKa using only simple and low-cost instrumentation. The optical part of the experimental setup has been optimized through the exclusion of the monochromator device. Thus it only takes 10-15 min to obtain one pKa value with the absolute error of less than 0.15 pH units. Application limitations and reliability of the method have been tested for a series of organic acids of various nature.

Topics: Acids; Hydrogen-Ion Concentration; Nitrophenols; Photometry; Reproducibility of Results; Spectrophotometry, Ultraviolet

This work concerns a comparison of experimental and theoretical results of the electron charge density distribution and the electrostatic potential around the m-nitrophenol molecule (m-NPH) known for its interesting physical characteristics. The molecular experimental results have been obtained from a high-resolution X-ray diffraction study. Theoretical investigations were performed using the Density Functional Theory at B3LYP level of theory at 6-31G* in the Gaussian program. The multipolar model of Hansen and Coppens was used for the experimental electron charge density distribution around the molecule, while we used the DFT methods for the theoretical calculations. The electron charge density obtained in both methods allowed us to find out different molecular properties such us the electrostatic potential and the dipole moment, which were finally subject to a comparison leading to a good match obtained between both methods. The intramolecular charge transfer has also been confirmed by an HOMO-LUMO analysis.

Topics: Crystallography, X-Ray; Hydrogen Bonding; Models, Molecular; Molecular Conformation; Nitrophenols; Quantum Theory; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis, Raman; Static Electricity; Thermodynamics; X-Ray Diffraction

Nitroaromatics are widely used for industrial purposes and constitute a group of compounds of environmental concern because of their persistence and toxic properties. Biological processes used for decontamination of nitroaromatic-polluted sources have then attracted worldwide attention. In the present investigation m-nitrophenol (MNP) biodegradation was studied in batch and continuous reactors. A bacterial community able to degrade the compound was first selected from a polluted freshwater stream and the isolates were identified by the analysis of the 16S rRNA gene sequence. The bacterial community was then used in biodegradation assays. Batch experiments were conducted in a 2L aerobic microfermentor at 28 °C and with agitation (200 rpm). The influence of abiotic factors in the biodegradation process in batch reactors, such as initial concentration of the compound and initial pH of the medium, was also studied. Continuous degradation of MNP was performed in an aerobic up-flow fixed-bed biofilm reactor. The biodegradation process was evaluated by determining MNP and ammonium concentrations and chemical oxygen demand (COD). Detoxification was assessed by Vibrio fischeri and Pseudokirchneriella subcapitata toxicity tests. Under batch conditions the bacterial community was able to degrade 0.72 mM of MNP in 32 h, with efficiencies higher than 99.9% and 89.0% of MNP and COD removals respectively and with concomitant release of ammonium. When the initial MNP concentration increased to 1.08 and 1.44 mM MNP the biodegradation process was accomplished in 40 and 44 h, respectively. No biodegradation of the compound was observed at higher concentrations. The community was also able to degrade 0.72 mM of the compound at pH 5, 7 and 9. In the continuous process biodegradation efficiency reached 99.5% and 96.8% of MNP and COD removal respectively. The maximum MNP removal rate was 37.9 gm(-3) day(-1). Toxicity was not detected after the biodegradation process.

Topics: Aliivibrio fischeri; Biodegradation, Environmental; Biofilms; Bioreactors; Chlorophyta; Gram-Negative Bacteria; Inactivation, Metabolic; Nitrophenols; RNA, Ribosomal, 16S; Water Pollutants, Chemical

A new organic-organic salt, 3-nitrophenol-1,3,5-triazine-2,4,6-triamine (2/1) (3-NPM) has been synthesized by slow evaporation technique at room temperature. Single crystal X-ray diffraction analysis reveals that 3-NPM crystallizes in orthorhombic system with centrosymmetric space group Pbca and the lattice parameters are a=15.5150(6) Å, b=12.9137(6) Å, c=17.8323(6) Å, α=β=γ=90° and V=3572.8(2)(Å)(3). The geometry, fundamental vibrational frequencies are interpreted with the aid of structure optimization and normal coordinate force field calculations based on density functional theory (DFT) B3LYP/6-311G(d,p) method. IR and Raman spectra of 3-NPM have been recorded and analyzed. The complete vibrational assignments are made on the basis of potential energy distribution (PED). The electric dipole moment, polarizability and the first order hyperpolarizability values of the 3-NPM have been calculated. (1)H and (13)C NMR chemical shifts are calculated by using the gauge independent atomic orbital (GIAO) method with B3LYP method with 6-311G (d,p) basis set. Moreover, molecular electrostatic potential (MEP) and thermodynamic properties are performed. Mulliken and Natural charges of the title molecule are also calculated and interpreted. Thermal decomposition behavior of 3-NPM has been studied by means of thermogravimetric analysis. The dielectric measurements on the powdered sample have been carried out and the variation of dielectric constant and dielectric loss at different frequencies of the applied field has been studied and the results are discussed in detail.

Topics: Magnetic Resonance Spectroscopy; Models, Molecular; Nitrophenols; Quantum Theory; Spectroscopy, Fourier Transform Infrared; Thermodynamics; Triazines

A 4-chloro-3-nitrophenol (4C3NP)-mineralizing bacterium, Pseudomonas sp. JHN was isolated from a waste water sample collected from a chemically-contaminated area, India by an enrichment method. Pseudomonas sp. JHN utilized 4C3NP as a sole carbon and energy source and degraded it with the release of stoichiometric amounts of chloride and nitrite ions. Gas chromatography and gas chromatography-mass spectrometry detected 4-chlororesorcinol as a major metabolite of the 4C3NP degradation pathway. Inhibition studies using 2,2'-dipyridyl showed that 4-chlororesorcinol is a terminal aromatic compound in the degradation pathway of 4C3NP. The activity for 4C3NP-monooxygenase was detected in the crude extracts of the 4C3NP-induced JHN cells that confirmed the formation of 4-chlororesorcinol from 4C3NP. The capillary assay showed that Pseudomonas sp. JHN exhibited chemotaxis toward 4C3NP. The bioremediation capability of Pseudomonas sp. JHN was monitored to carry out the microcosm experiments using sterile and non-sterile soils spiked with 4C3NP. Strain JHN degraded 4C3NP in sterile and non-sterile soil with same degradation rates. This is the first report of (i) bacterial degradation and bioremediation of 4C3NP, (ii) formation of 4-chlororesorcinol in the degradation pathway of 4C3NP, (iii) bacterial chemotaxis toward 4C3NP.

Topics: Biodegradation, Environmental; Chromatography, High Pressure Liquid; Humans; India; Metabolic Networks and Pathways; Nitrophenols; Pseudomonas; Resorcinols; Soil Microbiology; Wastewater

The use of cyclodextrins (CDs) for controlled delivery of drugs is largely presented in the literature. However, the question of whether CDs themselves linked to a polymeric network are able to sustain the release of drugs still persists. Here, CD immobilization within dextran microspheres is reported, and CD-dextran complexes were packed in a glass column and then, the retention time of different drugs and drug model compounds was determined by liquid chromatography. The release profiles of drugs and of drug model compounds (indole, 3-nitrophenol, p-hydroxybenzoic acid, diclofenac), characterized by different values of the retention time (high, moderate or low), were investigated. The release rates were quite high even for drugs that exhibit very high retention time (high association equilibrium constant). Moreover, the volume of the release fluid strongly influences the rate of drug release. As a whole, "the sink conditions" must be continuously maintained, since at each drug concentration in the release medium, equilibrium occurs between the free and the CD-bound drug.

Topics: alpha-Cyclodextrins; beta-Cyclodextrins; Chemistry, Pharmaceutical; Chromatography, Liquid; Cyclodextrins; Delayed-Action Preparations; Dextrans; Diclofenac; Drug Carriers; gamma-Cyclodextrins; Indoles; Kinetics; Microspheres; Models, Chemical; Nitrophenols; Parabens; Solubility; Technology, Pharmaceutical

Many biochromophore anions located within protein pockets display charge-transfer (CT) transitions that are perturbed by the nearby environment, such as water or amino acid residues. These anions often contain the phenolate moiety as the electron donor and an acceptor group that couples to the donor via a π-conjugated system. Here we show using action spectroscopy that single molecules of water, methanol, and acetonitrile cause blue shifts in the electronic transition energy of the bare m-nitrophenolate anion by 0.22, 0.22, and 0.12 eV, respectively (uncertainty of 0.05 eV). These shifts are similar to CC2-predicted ones and are in accordance with the weaker binding to the phenolate end of the ion by acetonitrile in comparison with water and methanol. The nitro acceptor group is almost decoupled from the phenolate donor, and this ion therefore represents a good model for CT excitations of an anion. We found that the shift caused by one acetonitrile molecule is almost half of that experienced in bulk acetonitrile solution, clearly emphasizing the important role played by the microenvironment. In protic solvents, the shifts are larger because of hydrogen bonds to the phenolate oxygen. Finally, but not least, we provide experimental data that serve to benchmark calculations of excited states of ion-solvent complexes.

Topics: Acetonitriles; Anions; Methanol; Molecular Structure; Nitrophenols; Solvents; Water

The effect of nitro-substituent on mononitrophenol (o-nitrophenol (ONP), m-nitrophenol (MNP) and p-nitrophenol (PNP)) reduction in a bioelectrochemical system (BES) was investigated in this study. The results show that the removal of all three nitrophenols was significantly enhanced with more negative cathode potential and shortened hydraulic retention time in the BESs. Moreover, the reduction of the three nitrophenols followed in the order of ONP > MNP > PNP in the BESs. Both quantum chemical calculation using density function theory and cyclic voltammetry analysis confirmed the reductive sequence of the three nitrophenols. In addition, the acute toxicity of nitrophenol effluent significantly decreased while its biodegradability was enhanced after treatment in the BES. Therefore, the BES technology offers bright prospects for efficient treatment of nitrophenol-containing wastewater.

Topics: Biodegradation, Environmental; Electrochemistry; Molecular Structure; Nitrophenols; Waste Disposal, Fluid; Wastewater

Pollutant interactions during the aerobic biodegradation of phenolic mixtures with either 2-nitrophenol (2-NP) or 3-nitrophenol (3-NP) by a NP-adapted microbial consortium in simulated wastewater were studied in a packed-bed bench scale bioreactor continuously operated in a flow mode. Phenol/2-NP and phenol/3-NP mixtures with varied phenol/nitrophenol ratios were shown to exhibit different biodegradability patterns. The presence of 2-NP led to a much lower overall elimination capacity and lower process stability in comparison to mixtures with 3-NP. In contrast to the expected greater degradation of a more biodegradable substrate in mixtures, phenol was degraded with a lower efficiency at higher phenol concentrations than NPs, although this difference became less pronounced with the gradual biofilm adaptation to phenol. This unusual substrate interaction, which appears to be common in the biotreatment of substituted phenol mixtures, was explained by prior biofilm adaptation to less degradable substrates, NPs. The biofilm composition was significantly altered during the long-term reactor operation. Although eukaryotes were not present in the inoculum, four fungal species were isolated from the biofilm after 1.5 years of operation. Of the initially present strains, only Chryseobacterium sp. and several Pseudomonas species persisted till the end of operation.

Topics: Bacterial Physiological Phenomena; Biodegradation, Environmental; Biofilms; Bioreactors; Chromatography, High Pressure Liquid; Nitrophenols; Phenol; Waste Disposal, Fluid; Wastewater; Water Pollutants, Chemical

The accelerated biodegradation of 3-nitrophenol (3-NP) in the rhizosphere of giant duckweed (Spirodela polyrrhiza) was investigated.. Biodegradation of 3-nitrophenol in the rhizosphere of a floating aquatic plant, S. polyrrhiza, was investigated by using three river water samples supplemented with 10 mg l(-1) of 3-NP. Isolation and enrichment culture of 3-NP-degrading bacteria were performed in basal salts medium containing 3-NP (50 mg l(-1)). The isolated strains were physiologically and phylogenetically characterized by using an API20NE kit and 16S rRNA gene sequencing.. Accelerated removal of 3-NP (100%) was observed in river water samples with S. polyrrhiza compared with their removal in plant-free river water. Also, 3-NP persisted in an autoclaved solution with aseptic plants, suggesting that the accelerated 3-NP removal resulted largely from degradation by bacteria inhabiting the plant rather than from adsorption and uptake by the plant. We successfully isolated six and four strains of 3-NP-degrading bacteria from the roots of S. polyrrhiza and plant-free river water, respectively. Phylogenetic analysis based on 16S rRNA gene divided the 3-NP-degrading bacteria into two taxonomic groups: the genera Pseudomonas and Cupriavidus. The strains belonging to the genus Cupriavidus were only isolated from the roots of duckweed. All strains isolated from the roots utilized 3-NP (0.5 mM) as a sole carbon and energy source, indicating that they could have contributed to the accelerated degradation of 3-NP in the rhizosphere of S. polyrrhiza.. The rhizoremediation using S. polyrrhiza and its rhizosphere bacteria can be an effective strategy for cleaning up the 3-NP-contaminated surface waters.

Topics: Araceae; Biodegradation, Environmental; Cupriavidus; Fresh Water; Molecular Sequence Data; Nitrophenols; Phylogeny; Pseudomonas; Rhizosphere; Rivers; RNA, Ribosomal, 16S

Four biodegradability tests (BOD(5)/COD ratio, production of carbon dioxide, relative oxygen uptake rate and relative enzymatic activity) were used to determine the aerobic biodegradability of 3-nitrophenol (3-NP), 2,4-dinitrophenol (2,4-DNP) and 2,6-dinitrophenol (2,6-DNP). Furthermore, biodegradation kinetics of the compounds was investigated in sequencing batch reactors both in the presence of glucose (co-substrate) and with nitrophenol as the sole carbon source. Among the three tested compounds, 3-NP showed the best biodegradability while 2,6-DNP was the most difficult to be biodegraded. The Haldane equation was applied to the kinetic test data of the nitrophenols. The kinetic constants are as follows: the maximum specific degradation rate (K(max)), the saturation constants (K(S)) and the inhibition constants (K(I)) were in the range of 0.005-2.98 mg(mgSS d)(-1), 1.5-51.9 mg L(-1) and 1.8-95.8 mg L(-1), respectively. The presence of glucose enhanced the degradation of the nitrophenols at low glucose concentrations. The degradation of 3-NP was found to be accelerated with the increasing of glucose concentrations from 0 to 660 mg L(-1). At high (1320-2000 mg L(-1)) glucose concentrations, the degradation rate of 3-NP was reduced and the K(max) of 3-NP was even lower than the value obtained in the absence of glucose, suggesting that high concentrations of co-substrate could inhibit 3-NP biodegradation. At 2,4-DNP concentration of 30 mg L(-1), the K(max) of 2,4-DNP with glucose as co-substrate was about 30 times the value with 2,4-DNP as sole substrate. 2,6-DNP preformed high toxicity in the case of sole carbon source degradation and the kinetic data was hardly obtained.

Topics: 2,4-Dinitrophenol; Aerobiosis; Biodegradation, Environmental; Biological Oxygen Demand Analysis; Glucose; Kinetics; Models, Theoretical; Nitrophenols; Wastewater; Water Pollutants, Chemical; Water Purification

SEIRA, SERS, TPD and DFT were used to study 4-nitrophenol (4NP), 3-nitrophenol (3NP) and 2-nitrophenol (2NP) adsorption on nanoscale silver films/powder. SERS and DFT demonstrated that 4NP adsorbed as the 4-nitrophenolate ion. SEIRA results revealed that a 4NP multilayer condensed differently using deposition solvents with and without polar bonds. 3NP and 2NP adsorption were not altered by the polar properties of the solvent. The nanoscale properties of the silver films/powder were shown to impact how the polar properties of the deposition solvent altered nitrophenol adsorption. In the SEIRA spectra of 4NP and 3NP a C=O stretch was observed above 1700cm(-1) using a highly volatile n-pentane deposition solvent. No other solvent yielded such a peak for 4NP or 3NP adsorption including n-heptane. 2NP had a C=O stretch regardless of deposition solvent. A C=O stretch confirmed nitrophenol ionization in the monolayer and pointed toward the significance of resonance in NP adsorption. 2NP never formed a multilayer at high exposures as demonstrated using TPD and SEIRA. Results of this work will have environmental implications and will aid biochemical and industrial applications where phenolic compounds are employed.

Topics: Adsorption; Isomerism; Models, Molecular; Nanostructures; Nitrophenols; Silver; Spectrum Analysis

In order to investigate nitro-substitutent's effect on degradation of phenols at boron-doped diamond (BDD) anodes, cyclic voltammetries of three nitrophenol isomers: 2-nitrophenol (2NP), 3-nitrophenol (3NP) and 4-nitrophenol (4NP) were studied, and their bulk electrolysis results were compared with phenol's (Ph) under alkaline condition. The voltammetric study showed nitrophenols could be attacked by hydroxyl radicals and nitro-group was released from the aromatic ring. Results of bulk electrolysis showed degradation of all phenols were fit to a pseudo first-order equation and followed in this order: 2NP>4NP>3NP>Ph. Molecular structures, especially carbon atom charge, significantly influenced the electrochemical oxidation of these isomers. Intermediates were analyzed during the electrolysis process, and were mainly catechol, resorcinol, hydroquinone, and carboxylic acids, such as acetic acid and oxalic acid. A simple degradation pathway was proposed. Moreover, a linear increasing relationship between degradation rates and Hammett constants of the studied phenols was observed, which demonstrated that electrochemical oxidation of these phenols was mainly initiated by electrophilic attack of hydroxyl radicals at BDD anodes.

Topics: Boron; Diamond; Electrochemical Techniques; Electrodes; Electrolysis; Environmental Restoration and Remediation; Hydroxyl Radical; Nitrophenols; Oxidation-Reduction; Phenols

Cupriavidus necator JMP134 utilizes meta-nitrophenol (MNP) as the sole source of carbon, nitrogen, and energy. The metabolic reconstruction of MNP degradation performed in silico suggested that MnpC might have played an important role in MNP degradation. In order to experimentally confirm the prediction, we have now characterized the mnpC-encoded (amino)hydroquinone dioxygenase involved in the ring-cleavage reaction of MNP degradation. Real-time PCR analysis indicated that mnpC played an essential role in MNP degradation. MnpC was purified to homogeneity as an N-terminal six-His-tagged fusion protein, and it was proved to be a dimer as demonstrated by gel filtration. MnpC was a Fe(2+)- and Mn(2+)-dependent dioxygenase, catalyzing the ring-cleavage of hydroquinone to 4-hydroxymuconic semialdehyde in vitro and proposed as an aminohydroquinone dioxygenase involved in MNP degradation in vivo. Phylogenetic analysis suggested that MnpC diverged from the other (chloro)hydroquinone dioxygenases at an earlier point, which might result in the preference for its physiological substrate.

Topics: Amino Acid Sequence; Carbon; Cloning, Molecular; Cupriavidus necator; Dioxygenases; Gene Expression; Genes, Bacterial; Hydroquinones; Nitrogen; Nitrophenols; Nitroreductases; Phylogeny; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Substrate Specificity

A three-phase hollow fiber liquid-phase microextraction method coupled with CE was developed and used for the determination of partition coefficients and analysis of selected nitrophenols in water samples. The selected nitrophenols were extracted from 14 mL of aqueous solution (donor solution) with the pH adjusted to pH 3 into an organic phase (1-octanol) immobilized in the pores of the hollow fiber and finally backextracted into 40.0 microL of the acceptor phase (NaOH) at pH 12.0 located inside the lumen of the hollow fiber. The extractions were carried out under the following optimum conditions: donor solution, 0.05 M H(3)PO(4), pH 3.0; organic solvent, 1-octanol; acceptor solution, 40 microL of 0.1 M NaOH, pH 12.0; agitation rate, 1050 rpm; extraction time, 15 min. Under optimized conditions, the calibration curves for the analytes were linear in the range of 0.05-0.30 mg/L with r(2)>0.9900 and LODs were in the range of 0.01-0.04 mg/L with RSDs of 1.25-2.32%. Excellent enrichment factors of up to 398-folds were obtained. It was found that the partition coefficient (K(a/d)) values were high for 2-nitrophenol, 3-nitrophenol, 4-nitrophenol, 2,4-dinitrophenol and 2,6-dinitrophenol and that the individual partition coefficients (K(org/d) and K(a/org)) promoted efficient simultaneous extraction from the donor through the organic phase and further into the acceptor phase. The developed method was successfully applied for the analysis of water samples.

Topics: 1-Octanol; 2,4-Dinitrophenol; Chromatography, Liquid; Electrophoresis, Capillary; Limit of Detection; Nitrophenols; Solvents; Uncoupling Agents; Water

The spectral shifts in the visible electronic absorption spectra of three amino-nitro-benzene derivatives in different solvents were correlated with the macroscopic parameters (refractive index and electric permittivity) of the solvents. The wavenumbers in the maximum of the visible charge transfer absorption band of o-nitro-aniline, 4-amino-3-nitrophenol and 3-amino-4-nitrophenol depend linearly on the Baur-Nicol function of electric permittivity. This dependence allows to estimate the electric polarizability in the electronic excited states if the electric polarizability in the ground state of the spectrally active molecule is determined by other procedures, such as by quanto-mechanical calculations.

Topics: Aniline Compounds; Benzene Derivatives; Coloring Agents; Humans; Molecular Structure; Nitrophenols; Solvents; Spectrum Analysis

Carbonaceous adsorbents prepared from olive stones biomass and novolac resin, as well as a commercial activated carbon for comparison reasons, have been examined for the removal of phenol and 3-nitrophenol from aqueous solutions. All carbonaceous adsorbents have been characterized by SEM-EDAX analysis and mercury porosimetry. The experimental data were examined according to the following kinetic models: pseudo first order, pseudo second order, Natarajan and Khalaf, Elovich, power function equations and intraparticle diffusion. By plotting the amount of adsorbate (phenol or 3-nitrophenol) adsorbed per unit mass of adsorbent b(t), versus the square root of time, four regions can be distinguished (A, B, C and D). By applying all the previously described models it is concluded that: (a) the intraparticle diffusion model is valid for the B and C region, whereas macropore diffusion and mesopore diffusion, respectively, take place. The pore diffusion coefficient, D for each carbonaceous material was calculated and indicated that the adsorption is controlled by diffusion, (b) the power model for the adsorption of phenol on each of the three carbonaceous materials is acceptable only for the C region and (c) the pseudo second order for the adsorption of 3-nitrophenol on each of the three carbonaceous materials is acceptable for the C region.

Topics: Adsorption; Carbon; Diffusion; Kinetics; Microscopy, Electron, Scanning; Nitrophenols; Phenol; Solutions; Spectrophotometry, Ultraviolet; Time Factors; Waste Disposal, Fluid; Water Pollutants, Chemical; Water Purification

Experiments have been performed with a semicontinuous batch reactor to compare the degradation efficiency of nitrobenzene in aqueous solution by the ultrasonic processes of single field, opposite dual fields, and orthogonal dual fields. Ultrasound with dual fields can improve the degradation efficiency of nitrobenzene compared to that of single field, and the improvement phenomenon is even more pronounced in the orthogonal dual-field system. The degradation reactions of nitrobenzene in the three processes all follow the pseudofirst-order kinetic model. The mechanism investigation indicates the degradation proceeds via hydroxyl radical (*OH) oxidation. The enhancement efficiency of orthogonal dual fields is attributed to an obvious synergetic effect, which accelerates the *OH initiation from 0.28 micromol L(-1) min(-1) for a single field to 0.98 micromol L(-1) min(-1) compared with 0.42 micromol L(-1) min(-1) for opposite dual fields, resulting in rapid formation of an increased diversity of byproducts and an advanced degree of mineralization of total organic carbon (TOC). The introduction of an ultrasonic field placed in the different spatial position causes a variable kinetic order during the removal of TOC. The degradation byproducts are identified by gas chromatography mass spectrometry and ion chromatography, including p-, m-nitrophenol, malonic acid, nitrate ion, 4-nitrocatechol, phenol, maleic acid, oxalic acid, hydroquinone, 1,2,3-trihydroxy-5-nitrobenzene, and acetic acid.

Topics: Acetic Acid; Carbon; Catechols; Chromatography, Gas; Hydroquinones; Hydroxyl Radical; Ions; Kinetics; Maleates; Malonates; Nitrates; Nitrobenzenes; Nitrophenols; Oxalic Acid; Ultrasonics; Water

Resolution of overlapped chromatographic peaks is generally accomplished by modeling the peaks as Gaussian or modified Gaussian functions. It is possible, even preferable, to use actual single analyte input responses for this purpose and a nonlinear least squares minimization routine such as that provided by Microsoft Excel Solver can then provide the resolution. In practice, the quality of the results obtained varies greatly due to small shifts in retention time. I show here that such deconvolution can be considerably improved if one or more of the response arrays are iteratively shifted in time.

Topics: Algorithms; Chromatography, Ion Exchange; Data Interpretation, Statistical; Nitrophenols; Normal Distribution; Software

We have previously characterized a number of small molecule organic compounds that prevent the aggregation of the beta-amyloid peptide and its neurotoxicity in hippocampal neuronal cultures. We have now evaluated the effects of such compounds on amyloid precursor protein (APP) accumulation in the CTb immortalized cell line derived from the cerebral cortex of a trisomy 16 mouse, an animal model of Down's syndrome. Compared to a non-trisomic cortical cell line (CNh), CTb cells overexpress APP and exhibit slightly elevated resting intracellular Ca2+ levels ([Ca2+] inverted exclamation mark). Here, we show that the compounds 2,4-dinitrophenol, 3-nitrophenol and 4-anisidine decreased intracellular accumulation of APP in CTb cells. Those compounds were non-toxic to the cells, and slightly increased the basal [Ca2+] inverted exclamation mark. Results indicate that the compounds tested can be leads for the development of drugs to decrease intracellular vesicular accumulation of APP in trisomic cells.

Topics: 2,4-Dinitrophenol; Alzheimer Disease; Amyloid beta-Protein Precursor; Aniline Compounds; Animals; Cell Line; Cerebral Cortex; Disease Models, Animal; Down Syndrome; Mice; Nitrophenols

A simple, novel and sensitive spectrophotometric method was described for simultaneous determination of nitrophenol isomers mixtures. All factors affecting on the sensitivity were optimized and the linear dynamic range for determination of nitrophenol isomers were found. The simultaneous determination of nitrophenol mixtures by using spectrophotometric methods is a difficult problem, due to the spectral interferences. The partial least squares modeling was used for the multivariate calibration of the spectrophotometric data. The orthogonal signal correction was used for preprocessing of data matrices and the prediction results of model, with and without using orthogonal signal correction, were statistically compared. The experimental calibration matrix was designed by measuring the absorbance over the range 300-520 nm for 21 samples of 1-20, 1-20 and 1-10 microg ml(-1) of m-nitrophenol, o-nitrophenol and p-nitrophenol, respectively. The RMSEP for m-nitrophenol, o-nitrophenol and p-nitrophenol with and without OSC were 0.3682, 0.5965, 0.3408 and 0.7351, 0.9962, 1.0055, respectively. The proposed method was successfully applied for the determination of m-nitrophenol, o-nitrophenol and p-nitrophenol in synthetic and real matrix samples such as water.

Topics: Industrial Waste; Least-Squares Analysis; Nitrophenols; Rivers; Spectrophotometry; Water Pollutants, Chemical; Water Supply

The removal efficiencies of 3-nitrophenol (3-NP) and 2, 6-dinitrophnol (2, 6-DNP) were investigated in two lab-scale upflow anaerobic sludge bed (UASB) reactors using two different co-substrates. Initially, glucose was used as co-substrate and followed by sodium acetate. The results showed that glucose was found to be a better co-substrate for 3-NP degradation compared to sodium acetate. While for the degradation of 2,6-dinitrophenol, sodium acetate was better. For the study of 3-NP degradation, input COD concentration was kept as 2,500 mg/L and hydraulic retention time (HRT) was kept as 26 h with glucose as co-substrate. Maximum 3-NP concentration was 254.6 mg/L and 3-NP removal efficiencies were always more than 99.0%. While HRT was 30 h with sodium acetate as co-substrate, maximum 3-NP concentration was 71.6 mg/L and over 90.0% 3-NP removal efficiencies could be obtained. For the study of 2,6-DNP degradation, HRT was 35 h using the same input COD concentration as 3-NP degradation. The maximum 2,6-DNP concentration was 170.0 mg/L and 2,6-DNP removal efficiencies were always more than 98.0% using glucose as co-substrate. While HRT was 30 h with sodium acetate as co-substrate, maximum 2,6-DNP concentration was 189.5 mg/L and over 99.2% 2,6-DNP removal efficiencies could be obtained.

Topics: Bioreactors; Carbon; Chemical Precipitation; Glucose; Nitrophenols; Sewage; Sodium Acetate; Waste Disposal, Fluid

Characteristics of anaerobic granules before and after acclimation, removal efficiencies of 3-nitrophenol (3-NP) and performance of 3-NP wastewater treatment were investigated in a lab-scale upflow anaerobic sludge bed (UASB). The results showed that granular sludge acclimatized to 3-NP rapidly during acclimation. Using SEM, the filamentous bacteria was the predominant bacteria on the surface of granules. Throughout the study of 3-NP wastewater anaerobic degradation, hydraulic retention time (HRT) and input COD concentration were kept constant as 26 h and 2 500 mg/L, respectively. While 3-NP concentrations increasing from 20 mg/L to 250 mg/L gradually, COD removal decreased from 95.2% to 85.1% and 3-NP removal efficiencies were always more than 99%. 3-AP was found to be the major intermediate during the degradation of 3-NP. Out of the total input of 3-NP concentration, on molar basis, about 58.7% - 111.9% of 3-NP was recovered in the form of 3-AP. With increasing 3-NP concentration, the variations of gas production were minimal. The minimum methane percentage was 65% and the maximum was 74%.

Topics: Bacteria, Anaerobic; Biodegradation, Environmental; Bioreactors; Methane; Nitrophenols; Sewage; Waste Disposal, Fluid

The bacterial strain J3 was isolated from soil by selective enrichment on mineral medium containing 4-nitrocatechol as the sole carbon and energy source. This strain was identified as Rhodococcus wratislaviensis on the basis of morphology, biochemical, physiological and chemotaxonomic characterization and complete sequencing of the 16S rDNA gene. The isolated bacterium could utilize 4-nitrocatechol, 3-nitrophenol and 5-nitroguaiacol as sole carbon and energy sources. Stoichiometric release of nitrites was measured during degradation of 4-nitrocatechol both in growing cultures and for stationary phase cells. The J3 strain was unable to degrade 4-nitroguaiacol, 2-nitrophenol, 4-nitrophenol, 2,4-dinitrobenzoic acid, 4,5-dimethoxy-2-nitrobenzoic acid and 2,3-difluoro-6-nitrophenol. The J3 strain is deposited in the Czech Collection of Microorganisms as CCM 4930.

Topics: Bacterial Typing Techniques; Catechols; DNA, Bacterial; DNA, Ribosomal; Fatty Acids; Genes, rRNA; Molecular Sequence Data; Mycolic Acids; Nitrites; Nitro Compounds; Nitrophenols; Phylogeny; Rhodococcus; RNA, Bacterial; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology

Concentration-dependent inhibition of the photosynthetic electron transport chain (photosystem I (PS I), photosystem II (PS II) and whole chain reaction) and ATP content was observed in Nostoc muscorum and Chlorella vulgaris grown with o-nitrophenol, m-nitrophenol, or 2,4-dinitrophenol. Although the extents of inhibition of the photosynthetic electron transport chain in both organisms were similar, PS II was more sensitive than PS I and whole chain reaction to the nitrophenols. Depletion of the ATP pool was noted in nitrophenol-grown cultures, probably as a consequence of nearly complete inhibition of the photosynthetic electron transport chain.

Topics: 2,4-Dinitrophenol; Adenosine Triphosphate; Biodegradation, Environmental; Chlorella; Culture Media; Cyanobacteria; Electron Transport; Nitrophenols; Photosynthesis; Photosystem I Protein Complex; Photosystem II Protein Complex; Uncoupling Agents

We report on the effect on performance of varying the length of the capillary during throughout in-capillary derivatization (TICD) capillary electrophoresis (CE). Performance was evaluated by on-line coupling with a sample and CE runbuffer loading device that was newly introduced for this study. The device was assembled with a low cost using two 5 mm inner diameter (ID) disposable polyethylene syringes. First, a sequence was manually formed consisting of a 200 microL run buffer solution plug, a 100 microL sample plug and another 200 microL run buffer solution plug. Each plug was separated from its neighbor by a 100 microL air plug. When each plug reached the injection point where both a platinum-wire anode and the end of the separation capillary tube were located, 340 V/cm separation voltage (electrophoresis voltage) and 34 V/cm injection voltage were applied to the capillary for 3 s. Then the analytes were derivatized during migration in 50 microm ID capillaries filled with 2 mM o-phthalaldehyde (OPA)/N-acetylcysteine (NAC) in a 20 mM phosphate-borate buffer (pH 10), followed by separating and detecting of OPA derivatives by absorbance of 340 nm. Derivatization, separation, and detection were performed systematically using capillaries which varied in length from 5 to 80 cm. In the case of TICD-CE of a mixture containing 1 mM aspartic acid (Asp) and 20 mM m-nitorophenol (MNP) as a test solution, it was determined that peak area and peak width ratios of Asp to MNP did not depend on capillary length. Enantiomeric separations of DL-alanine (Ala) and Asp were examined using a run buffer consisting of a 45 microM beta-cyclodextrin (CD)-2 mM OPA/NAC-20 mM phosphate-borate buffer (pH 10). Even though the resolution of these enantiomeric pairs decreased with decreasing capillary length, as expected, the peaks corresponding to both enantiomeric amino acids were identified even when a 5 cm capillary was used. An 8-component amino acid mixture was also tested with 5 cm and 10 cm capillaries.

Topics: Aspartic Acid; Buffers; Electrophoresis, Capillary; Nitrophenols; o-Phthalaldehyde

Hollow chitosan fibers were reacted with chloropalladate solutions and subsequently reduced by hydrogen produced in situ by reaction of sulfuric acid with zinc powder in order to manufacture palladium supported on catalytic hollow chitosan fibers (C2HF-Pd). This catalytic support was used to degrade 3-nitrophenol (3-NP) using two different hydrogen donors (hydrogen gas and sodium formate). The solution was flowed through the lumen of the fiber, while the sodium formate was recirculated round the outside of the fiber. In the case of hydrogen gas, the gas was maintained under controlled pressure outside the fiber. The influence of the pH, residence time (ca. flow velocity), nitrophenol concentration, and hydrogen-donor concentration (or pressure) was investigated for both systems in order to evaluate the limiting parameters. While the system using sodium formate was the most efficient for nitrophenol conversion, the system using hydrogen gas avoided the production of secondary waste solutions (formate solutions with traces of nitrophenol, which pass through the fiber membrane).

Topics: Catalysis; Chitosan; Formates; Hydrogen; Hydrogen-Ion Concentration; Industrial Waste; Microscopy, Electron, Scanning; Nitrophenols; Palladium; Waste Management

The 3-nitrophenol-induced enzyme system in cells of Pseudomonas putida 2NP8 manifested a wide substrate range in transforming nitroaromatic compounds through to ammonia production. All of the 30 mono- or dinitroaromatic substrates except 4-nitrophenol, 2,4-dinitrophenol, 2,4,6-trinitrophenol, 3-nitroaniline, 2-nitrobenzoic acid, and 2-nitrofuran were quickly transformed. Ammonia production from most nitroaromatic substrates appeared to be stoichiometric.

Topics: Aerobiosis; Ammonia; Hydrocarbons, Aromatic; Nitrobenzenes; Nitrophenols; Nitroreductases; Pseudomonas putida; Substrate Specificity

In this work the peak suppression technique is used for the determination of 3-nitrophenol and some other aromatic impurities in 4-nitrophenol by reversed phase HPLC with diode array detection. Taking into account the differences between the absorption spectra of the two compounds, two wavelengths were selected in order to obtain the maximum difference between the spectral contribution for 3-nitrophenol and to maintain a small, similar spectral contribution for 4-nitrophenol (the main compound). Then we used the wavelength corresponding to a small spectral contribution of 3-nitrophenol as the reference wavelength. It was shown that taking lambda(an) = 266 nm and lambda(ref) = 364 nm, a broad elution peak of 4-nitrophenol was suppressed deconvoluting the peak of 3-nitrophenol. Moreover, quantitation of 3-nitrophenol was achieved without chemometric tools. Under the proposed conditions the detection limits for 3-nitrophenol and other common impurities of 4-nitrophenol used in the pharmaceutical industry (4-chlorophenol, 4-nitrophenol, 1-chloro-2-nitrobenzene, 1-chloro-4-nitrobenzene, 4,4'-bisfenilether, and 4,4'-dichloroazobenzene) were not significantly affected as compared with respective detection limits evaluated in the absence of 4-nitrophenol and using standard detection conditions (lambda(an) = 280 nm and lambda(ref) = 420 nm).

Topics: Chromatography, High Pressure Liquid; Nitrophenols; Sensitivity and Specificity; Spectrophotometry, Ultraviolet

Biotransformation products of hydroxylaminobenzene and aminophenol produced by 3-nitrophenol-grown cells of Pseudomonas putida 2NP8, a strain grown on 2- and 3-nitrophenol, were characterized. Ammonia, 2-aminophenol, 4-aminophenol, 4-benzoquinone, N-acetyl-4-aminophenol, N-acetyl-2-aminophenol, 2-aminophenoxazine-3-one, 4-hydroquinone, and catechol were produced from hydroxylaminobenzene. Ammonia, N-acetyl-2-aminophenol, and 2-aminophenoxazine-3-one were produced from 2-aminophenol. All of these metabolites were also found in the nitrobenzene transformation medium, and this demonstrated that they were metabolites of nitrobenzene transformation via hydroxylaminobenzene. Production of 2-aminophenoxazine-3-one indicated that oxidation of 2-aminophenol via imine occurred. Rapid release of ammonia from 2-aminophenol transformation indicated that hydrolysis of the imine intermediate was the dominant reaction. The low level of 2-aminophenoxazine-3-one indicated that formation of this compound was probably due to a spontaneous reaction accompanying oxidation of 2-aminophenol via imine. 4-Hydroquinone and catechol were reduction products of 2- and 4-benzoquinones. Based on these transformation products, we propose a new ammonia release pathway via oxidation of aminophenol to benzoquinone monoimine and subsequent hydrolysis for transformation of nitroaromatic compounds by 3-nitrophenol-grown cells of P. putida 2NP8. We propose a parallel mechanism for 3-nitrophenol degradation in P. putida 2NP8, in which all of the possible intermediates are postulated.

Topics: Aminophenols; Biotransformation; Chromatography, High Pressure Liquid; Culture Media; Hydroxylamines; Nitrophenols; Oxidation-Reduction; Pseudomonas putida; Time Factors

A strain of Pseudomonas putida (2NP8) capable of growing on both 2-nitrophenol and 3-nitrophenol, but not on nitrobenzene (NB), was isolated from municipal activated sludge. 2-Nitrophenol was degraded by this strain with production of nitrite. Degradation of 3-nitrophenol resulted in the formation of ammonia. Cells grown on 2-nitrophenol did not degrade nitrobenzene. A specific nitrobenzene degradation activity was induced by 3-nitrophenol. Ammonia, nitrosobenzene, and hydroxylaminobenzene have been detected as metabolites of nitrobenzene degradation by cells grown in the presence of 3-nitrophenol. These results indicated a NB cometabolism mediated by 3-nitrophenol nitroreductase.

Topics: Biodegradation, Environmental; Environmental Microbiology; Nitrobenzenes; Nitrophenols; Pseudomonas putida; Sewage

3-Hydroxylaminophenol mutase from Ralstonia eutropha JMP134 is involved in the degradative pathway of 3-nitrophenol, in which it catalyzes the conversion of 3-hydroxylaminophenol to aminohydroquinone. To show that the reaction was really catalyzed by a single enzyme without the release of intermediates, the corresponding protein was purified to apparent homogeneity from an extract of cells grown on 3-nitrophenol as the nitrogen source and succinate as the carbon and energy source. 3-Hydroxylaminophenol mutase appears to be a relatively hydrophobic but soluble and colorless protein consisting of a single 62-kDa polypeptide. The pI was determined to be at pH 4.5. In a database search, the NH2-terminal amino acid sequence of the undigested protein and of two internal sequences of 3-hydroxylaminophenol mutase were found to be most similar to those of glutamine synthetases from different species. Hydroxylaminobenzene, 4-hydroxylaminotoluene, and 2-chloro-5-hydroxylaminophenol, but not 4-hydroxylaminobenzoate, can also serve as substrates for the enzyme. The enzyme requires no oxygen or added cofactors for its reaction, which suggests an enzymatic mechanism analogous to the acid-catalyzed Bamberger rearrangement.

Topics: Amino Acid Sequence; Bacteria; Chromatography, Affinity; Chromatography, DEAE-Cellulose; Cupriavidus necator; Electrophoresis, Polyacrylamide Gel; Glutamate-Ammonia Ligase; Hydrogen-Ion Concentration; Intramolecular Transferases; Kinetics; Models, Chemical; Molecular Sequence Data; Molecular Weight; Nitrophenols; Sequence Alignment; Sequence Homology, Amino Acid; Substrate Specificity; Ultracentrifugation

The substrate specificities and product inhibition patterns of haloalkane dehalogenases from Xanthobacter autotrophicus GJ10 (XaDHL) and Rhodococcus rhodochrous (RrDHL) have been compared using a pH-indicator dye assay. In contrast to XaDHL, RrDHL is efficient toward secondary alkyl halides. Using steady-state kinetics, we have shown that halides are uncompetitive inhibitors of XaDHL with 1, 2-dichloroethane as the varied substrate at pH 8.2 (Cl-, Kii = 19 +/- 0.91; Br-, Kii = 2.5 +/- 0.19 mM; I-, Kii = 4.1 +/- 0.43 mM). Because they are uncompetitive with the substrate, halide ions do not bind to the free form of the enzyme; therefore, halide ions cannot be the last product released from the enzyme. The Kii for chloride was pH dependent and decreased more than 20-fold from 61 mM at pH 8.9 to 2.9 mM at pH 6.5. The pH dependence of 1/Kii showed simple titration behavior that fit to a pKa of approximately 7.5. The kcat was maximal at pH 8.2 and decreased at lower pH. A titration of kcat versus pH also fits to a pKa of approximately 7.5. Taken together, these data suggest that chloride binding and kcat are affected by the same ionizable group, likely the imidazole of a histidyl residue. In contrast, halides do not inhibit RrDHL. The Rhodococcus enzyme does not contain a tryptophan corresponding to W175 of XaDHL, which has been implicated in halide ion binding. The site-directed mutants W175F and W175Y of XaDHL were prepared and tested for halide ion inhibition. Halides do not inhibit either W175F or W175Y XaDHL.

Topics: Amino Acid Sequence; Binding, Competitive; Chlorides; Enzyme Inhibitors; Ethylene Chlorohydrin; Gram-Negative Aerobic Bacteria; Hydrogen-Ion Concentration; Hydrolases; Indicators and Reagents; Kinetics; Molecular Sequence Data; Mutagenesis, Site-Directed; Nitrophenols; Phenolsulfonphthalein; Phenylalanine; Rhodococcus; Substrate Specificity; Tryptophan; Tyrosine

Ralstonia eutropha JMP134 utilizes 2-chloro-5-nitrophenol as a sole source of nitrogen, carbon, and energy. The initial steps for degradation of 2-chloro-5-nitrophenol are analogous to those of 3-nitrophenol degradation in R. eutropha JMP134. 2-Chloro-5-nitrophenol is initially reduced to 2-chloro-5-hydroxylaminophenol, which is subject to an enzymatic Bamberger rearrangement yielding 2-amino-5-chlorohydroquinone. The chlorine of 2-amino-5-chlorohydroquinone is removed by a reductive mechanism, and aminohydroquinone is formed. 2-Chloro-5-nitrophenol and 3-nitrophenol induce the expression of 3-nitrophenol nitroreductase, of 3-hydroxylaminophenol mutase, and of the dechlorinating activity. 3-Nitrophenol nitroreductase catalyzes chemoselective reduction of aromatic nitro groups to hydroxylamino groups in the presence of NADPH. 3-Nitrophenol nitroreductase is active with a variety of mono-, di-, and trinitroaromatic compounds, demonstrating a relaxed substrate specificity of the enzyme. Nitrosobenzene serves as a substrate for the enzyme and is converted faster than nitrobenzene.

Topics: Biodegradation, Environmental; Cupriavidus necator; Nitro Compounds; Nitrophenols; Nitroreductases; Nitroso Compounds; Oxidation-Reduction; Substrate Specificity

Growth of Pseudomonas putida B2 in chemostat cultures on a mixture of 3-nitrophenol and glucose induced 3-nitrophenol and 1,2,4-benzenetriol-dependent oxygen uptake activities. Anaerobic incubations of cell suspensions with 3-nitrophenol resulted in complete conversions of the substrate to ammonia and 1,2,4-benzenetriol. This indicates that P. putida B2 degrades 3-nitrophenol via 1,2,4-benzenetriol, via a pathway involving a hydroxylaminolyase. Involvement of this pathway in nitroaromatic metabolism has previously only been found for degradation of 4-nitrobenzoate. Reduction of 3 nitrophenol by cell-free extracts was strictly NADPH-dependent. Attempts to purify the enzymes responsible for 3-nitrophenol metabolism were unsuccessful, because their activities were extremely unstable. 3-Nitrophenol reductase was therefore characterized in cell-free extracts. The enzyme had a sharp pH optimum at pH 7 and a temperature optimum at 25 degrees C. At 30 degrees C, reductase activity was completely destroyed within one hour, while at 0 degrees C, the activity in cell-free extracts was over 100-fold more stable. The Km values for NADPH and 3-nitrophenol were estimated at 0.17 mM and below 2 microM, respectively. The substrate specificity of the reductase activity was very broad: all 17 nitroaromatics tested were reduced by cell-free extracts. However, neither intact cells nor cell-free extracts could convert a set of synthesized hydroxylaminoaromatic compounds to the corresponding catechols and ammonia. Apparently, the hydroxylaminolyase of P. putida B2 has a very narrow substrate specificity, indicating that this organism is not a suitable biocatalyst for the industrial production of catechols from nitroaromatics.

Topics: Biodegradation, Environmental; Catechols; Cell-Free System; Culture Media; Enzyme Inhibitors; Hydrogen-Ion Concentration; Hydroquinones; Kinetics; Nitrophenols; Nitroreductases; Oxygen Consumption; Pseudomonas putida; Substrate Specificity; Temperature

The effective concentrations (EC), which cause a certain percentage of inhibition in growth yield, of four nitrophenolics (o-nitrophenol, m-nitrophenol, p-nitrophenol (PNP), and 2,4-dinitrophenol) and the major metabolites of PNP reduction (p-nitrosophenol and p-aminophenol) toward growth of two microalgae (Chlorella vulgaris and Scenedesmus bijugatus) and two cyanobacteria (Nostoc muscorum and Nostoc linckia), all isolated from soil, were determined following linear regression analysis. The EC50 values of the selected toxicants toward the test organisms were in the range of 32 to 227 micrograms ml-1. Such a wide range in EC figures emphasizes the need for a prior determination of EC values of a toxicant before establishing its toxicity pattern toward a nontarget microorganism.

Topics: 2,4-Dinitrophenol; Chlorella; Cyanobacteria; Eukaryota; Lethal Dose 50; Linear Models; Nitrophenols; Soil Microbiology; Structure-Activity Relationship

We have isolated two soil bacteria (identified as Arthrobacter aurescens TW17 and Nocardia sp. strain TW2) capable of degrading p-nitrophenol (PNP) and numerous other phenolic compounds. A. aurescens TW17 contains a large plasmid which correlated with the PNP degradation phenotype. Degradation of PNP by A. aurescens TW17 was induced by preexposure to PNP, 4-nitrocatechol, 3-methyl-4-nitrophenol, or m-nitrophenol, whereas PNP degradation by Nocardia sp. strain TW2 was induced by PNP, 4-nitrocatechol, phenol, p-cresol, or m-nitrophenol. A. aurescens TW17 initially degraded PNP to hydroquinone and nitrite. Nocardia sp. strain TW2 initially converted PNP to hydroquinone or 4-nitrocatechol, depending upon the inducing compound.

Topics: Arthrobacter; Biodegradation, Environmental; Catechols; Cresols; Gene Expression Regulation, Bacterial; Nitrophenols; Nocardia; Phenol; Phenols; Soil Microbiology; Soil Pollutants

The nipecotic acid ester m-nitrophenyl-3-piperidinecarboxylate (MNPC) possesses anticonvulsant activity. In the present study, the metabolites m-nitrophenol and nipecotic acid were determined in mouse blood and brain tissue after administration of MNPC. This determination was used as an indication of the distribution of the parent compound MNPC and to provide information regarding the differences in distribution between the enantiomers of MNPC, the times of onset, and effectiveness when (+/-) MNPC was administered by subcutaneous (sc) and intraperitoneal (ip) routes. m-Nitrophenol was determined by a previously reported high-performance liquid chromatography (HPLC) method. There was no significant difference in m-nitrophenol distribution after sc administration of (+)MNPC and (-)MNPC (400 mg/kg each). This similar pattern of distribution is in agreement with the earlier reported equi-effectiveness of the enantiomers of anticonvulsants. Peak m-nitrophenol levels in blood, which occurred at 15 min, were three times greater when (+/-) MNPC was administered by ip injection as compared with sc injection. This significant difference is most likely due to enhanced absorption and the peripheral metabolism of MNPC by the liver when the ip route is employed. A novel HPLC assay for the determination of nipecotic acid in mouse brain was developed, based on a modification of a reported amino acid analysis procedure. The results of the brain distribution studies showed that nipecotic acid brain levels peaked at 30 min after sc administration of (-)MNPC.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Anticonvulsants; Chromatography, High Pressure Liquid; Injections, Intraperitoneal; Injections, Subcutaneous; Male; Mice; Mice, Inbred Strains; Nipecotic Acids; Nitrophenols; Tissue Distribution

Topics: Acetylcholine; Acetylthiocholine; Animals; Brain; Cholinesterases; Electrophorus; Hydrolysis; Male; Mice; Nitrophenols; Rats; Rats, Inbred Strains; Spectrophotometry

The effects of the three hydrophobic molecules triphenylphosphine, trifluoperazine and 3-nitrophenol on Ca2+ uptake and ATPase activity in sarcoplasmic reticulum vesicles was investigated. When ATP was the substrate, triphenylphosphine (3 microM) increased the amount of Ca2+ accumulated by the vesicles. At high concentrations triphenylphosphine inhibited Ca2+ uptake. This effect varied depending on the ATP concentration and the type of nucleotide used. With ITP there was only inhibition and no activation of Ca2+ uptake by triphenylphosphine. On the other hand, trifluoperazine inhibited Ca2+ accumulation regardless of whether ATP or ITP was used as substrate. When 5 mM oxalate was included in the medium in order to avoid binding of Ca2+ to the low-affinity Ca2(+)-binding sites of the enzyme, both stimulation by triphenylphosphine and inhibition by trifluoperazine were reduced. In leaky vesicles at low Ca2+ concentrations, triphenylphosphine and 3-nitrophenol were competitive inhibitors of ATPase activity at the regulatory site of the enzyme (0.1-1 mM ATP). A striking difference was observed when both the high- and low-affinity Ca2(+)-binding sites were saturated. In this condition, triphenylphosphine and 3-nitrophenol promoted a 3-4-fold increase in the apparent affinity for ATP at its regulatory site.

Topics: Adenosine Triphosphate; Animals; Ca(2+) Mg(2+)-ATPase; Calcium; Calcium-Transporting ATPases; Kinetics; Models, Theoretical; Muscles; Nitrophenols; Organophosphorus Compounds; Oxalates; Protein Binding; Rabbits; Sarcoplasmic Reticulum; Trifluoperazine

Frogs injected with 3-nitrophenol excreted 85-93% of the administered dose within 17 h; 70-90% dose was metabolized. Metabolites identified comprise 3-nitrophenyl glucuronide (57% dose), 3-nitrophenyl sulphate (24% dose), and 3-acetamidophenyl sulphate (2% dose). Traces of the following metabolites were found: 3-acetamidophenyl glucuronide, 3-acetamidophenol, 4-nitrocatechol, nitroquinol, 4-nitrocatechol sulphate and nitroquinol sulphate.

Topics: Animals; Biotransformation; Chromatography, High Pressure Liquid; Hydrolysis; Nitrophenols; Rana temporaria

The ventilatory effects of the three mononitrophenols and six dinitrophenols have been examined in anaesthetized rats. The minute volume of ventilation increased in all the test groups, the increase reaching the 99% significance level with seven compounds (P<0.01), the 95% level with 3-nitrophenol (P<0.05), and the 90% level with 3,5-dinitrophenol (P<0.10). The effects of 4-nitrophenol, 3,4-dinitrophenol, 3-nitrophenol, and 2,5-dinitrophenol, in increasing carbon dioxide output relative to oxygen consumption, are not explicable on the basis of simple hyperventilation, and are attributed to a metabolic effect at the cellular level. The potency of the nitrophenols in stimulating respiration is related to their structure, nitro groups being most effective in the ortho position and least effective in the meta position, and 2,4-dinitrophenol being the most powerful respiratory stimulant of the group. 2,6-Dinitrophenol does not conform to this generalization; one unique feature of its structure is indicated, as a possible explanation for the discrepancy. The gradation of potency of the nitrophenols (except 2,6-dinitrophenol) parallels the gradation of acidic properties in the group; the more strongly acid compounds stimulate respiration more powerfully. This is not a direct effect on blood pH, since the compounds were administered in neutral or slightly alkaline solution. Methaemoglobin formation was found to occur with 2,5-dinitrophenol, and to a smaller inconstant extent with three other compounds. Further work is suggested, to explore whether peripheral-acting ventilatory stimulation by 2,4-dinitrophenol is necessarily associated with peripheral metabolic enhancement, or whether the two effects can be dissociated.

Topics: 2,4-Dinitrophenol; Animals; Cell Respiration; Dinitrophenols; Nitrophenols; Oxygen Consumption; Pulmonary Ventilation; Rats; Respiration

The effect of the mono- and di-nitrophenols and certain related compounds has been determined on the rate of oxygen consumption, the rate of carbon dioxide output and the rectal temperature of the Wistar albino rat.Of the compounds examined, only 2:4-dinitrophenol and its derivative, 3:5-dinitro-o-cresol, stimulated metabolic rate. 2-Nitrophenol and 2:3-, 2:6-, and 3:5-dinitrophenol produced no change in metabolic rate; 3-nitrophenol and 2:5-dinitrophenol had no action on carbon dioxide production although they caused a decrease in oxygen consumption. 4-Nitrophenol and 3:4-dinitrophenol increased only the rate of carbon dioxide output; 2-amino-4-nitrophenol increased the rate of carbon dioxide output and decreased the rate of oxygen consumption; 4-amino-2-nitrophenol caused depression of metabolic rate.It was confirmed that neither rectal temperature nor carbon dioxide output could replace rate of oxygen consumption as a reliable index of metabolic stimulant action. An apparatus is described which facilitates measurement of the oxygen consumption of small mammals.

Topics: Animals; Carbon Dioxide; Cresols; Dinitrophenols; Metabolism; Nitrophenols; Oxygen Consumption; Rats; Rats, Wistar