Interaction of tebuconazole with bovine serum albumin: determination of the binding mechanism and binding site by spectroscopic methods

Abstract

This study investigates the interaction between tebuconazole and bovine serum albumin (BSA) in a physiological buffer (pH = 7.4) using the fluorescence quenching method to obtain the apparent binding constants (K) and number of binding sites (n) in the interaction between tebuconazole and BSA. The results revealed that tebuconazole can quench the intrinsic fluorescence of BSA through a static quenching procedure. It also shows that the thermodynamic parameters of enthalpy change (ΔH) and entropy change (ΔS) are negative, indicating that the interaction of tebuconazole with BSA is mainly driven by van der Waals forces and hydrogen bonds. The process of binding was a spontaneous process in which Gibbs free energy change was negative. The distance of r between the donor (BSA) and acceptor (tebuconazole) was calculated to be 0.68?nm based on Forster’s non-radiative energy transfer theory. Analysis of synchronous fluorescence, three-dimensional fluorescence and circular dichroism (CD) spectra demonstrates that tebuconazole can induce conformational changes of BSA.     

Effects of increased sea water concentrations of CO2 on growth of the bivalve Mytilus edulis L

It has been proposed that emission of anthropogenic carbon dioxide to the atmosphere will lead to increased concentrations of CO(2) in sea water corresponding to a decrease of pH of several tenths of pH units. An experiment was performed to test the effects of increased sea water concentrations of CO(2) on shell growth of the blue mussel Mytilus edulis. The experiment was performed in aquaria continuously flushed with sea water spiked with CO(2) to provide five different levels of pH between 6.7 and control sea water with a pH of 8.1. The shell length of the mussels was measured at the start and end of the 44 days experimental period. No mortality was observed during the first 23 days of the experiment. The growth increment in mm was much larger for small mussels than for large mussels, but relative growth profile as function of pH was more similar in the two size groups; observed differences may be random variation between samples. The experiments showed that CO(2) induced reduction of pH affects the growth of M. edulis negatively. There was a strong and statistically significant decrease in growth at the lowest pH values, with virtually no growth at pH = 6.7 and reduced growth at pH = 7.1. The effect seems to set in between pH 7.4 and 7.1; at mean pH levels 7.4 and 7.6 the growth increments were not significantly different from growth at normal pH 8.1.     

Removal of humic substances from water by means of calcium-ion-enriched natural zeolites.

The ability of the natural zeolited Neapolitan Yellow Tuff (NYT) enriched with calcium ions to remove humic acids from water was evaluated by batch adsorption equilibrium tests and dynamic experiments carried out by percolating humic acid solutions through a small NYT column (breakthrough curves). Under the experimental condition explored, the sorption capacity increases with the ionic strength and has the highest value at pH 7.4. The partition coefficient for a low concentration of humic acid ([humic acid] --> 0), at pH 7.4 in 0.01 M sodium chloride, was approximately 1000 L/kg, versus the value of approximately 100 L/kg in the absence of the alkaline metal salt. Therefore, after humic acids have been adsorbed in a column filled with the calcium-ion-enriched tuff, a reduction of the salt concentration in the ongoing solution enhances the release of the adsorbed material. These findings show that NYT can be used for the removal of humic acids from water.     

Acid–base balance and metabolic response of the sea urchin Paracentrotus lividus to different seawater pH and temperatures

Purpose

In order to better understand if the metabolic responses of echinoids could be related to their acid?Cbase status in an ocean acidification context, we studied the response of an intertidal sea urchin species, Paracentrotus lividus, submitted to low pH at two different temperatures.

Methods

Individuals were submitted to control (8.0) and low pH (7.7 and 7.4) at 10°C and 16°C (19?days). The relation between the coelomic fluid acid?Cbase status, the RNA/DNA ratio of gonads and the individual oxygen uptake were studied.

Results

The coelomic fluid pH decreased with the aquarium seawater, independently of temperature, but this explained only 13% of the pH variation. The coelomic fluid showed though a partial buffer capacity that was not related to skeleton dissolution ([Mg²⁺] and [Ca²⁺] did not differ between pH treatments). There was an interaction between temperature and pH on the oxygen uptake (V _O2) which was increased at pH?7.7 and 7.4 at 10°C in comparison with controls, but not at 16°C, indicating an upregulation of the metabolism at low temperature and pH. However, gonad RNA/DNA ratios did not differ according to pH and temperature treatments, indicating that even if maintenance of physiological activities has an elevated metabolic cost when individuals are exposed to stress, they are not directly affected during short-term exposure. Long-term studies are needed in order to verify if gonad production/growth will be affected by low pH seawaters exposure.     

Chromium adsorption by aligned carbon nanotubes supported ceria nanoparticles

Ceria nanoparticles supported on aligned carbon nanotubes (CeO(2)/ACNTs), a novel adsorbent for Cr(VI) from drinking water, were prepared by chemical reaction of CeCl(3) with NaOH in aligned carbon nanotube solution and subsequent heat treatment. The best Cr(VI) adsorption effect of CeO(2)/ACNTs occurs at a pH range of 3.0-7.4. The largest adsorption capacity of CeO(2)/ACNTs reaches 30.2 mg g(-1) at an equilibrium Cr(VI) concentration of 35.3 mg l(-1) at pH 7.0. The experiment results suggest that CeO(2)/ACNTs have great potential applications in environmental protection.     

Adsorption of fulvic acid by carbon nanotubes from water

This study investigated adsorption of fulvic acid (FA) by single-walled (SWCNT) and multi-walled carbon nanotubes (MWCNT) and activated carbon. Adsorption of FA depends greatly on the adsorbent surface area and solution pH. SWCNT has higher adsorption than MWCNT and activated carbon. Lower E4/E6 (absorbance at 465 nm to that at 665 nm) and higher E2/E3 (absorbance at 250 nm to that at 365 nm) ratios of the residual FA in solution after adsorption than that of original FA in low pH ranges suggest that aromatic rich FA fractions with polar moieties readily adsorb on the adsorbents. The apparent interaction mechanisms between FA and CNT surfaces include electrostatic, hydrophobic, π-π and hydrogen-bond interactions. FA adsorption was reduced greatly with increasing pH because of the increase of electrostatic repulsion and the decrease of hydrophobic and hydrogen-bond interactions.     

Ionic strength effects in biosorption of metals by marine algae

Biosorption, the passive accumulation of metals by biomass, can be used as a cost-effective process for the treatment of metal polluted industrial effluents. The green alga Ulva fascia and the brown seaweeds Sargassum hemiphyllum, Petalonia fascia, and Colpomenia sinuosa were characterized in terms of their number of binding sites, their charge density and intrinsic proton binding constant (pKa) using pH titrations at different ionic strengths. The determined number of binding sites decreased in the order Petalonia > or = Sargassum > Colpomenia > Ulva. Due to their high number of binding sites Sargassum and Petalonia are most promising for biosorption applications. The decrease of proton binding with increasing ionic strength and pH as well as the increase of Cu and Ni binding with increasing pH and decreasing ionic strength could be described by the Donnan model in conjunction with an ion exchange biosorption isotherm.     

Complexing capacity of natural waters carrying a great amount of suspended matter

The cadmium binding properties of waters of the superior section of the Rio de la Plata estuarine were determined over a three-year period. Samples were collected at different hydrodynamic conditions. The complexing capacity was determined by square wave anodic stripping voltammetry (SWASV). Titration curve data were analyzed using a multivariable regression. Suspended particulate matter (SPM) was identified by XR diffraction and FTIR. These analyses showed that SPM principal components are clays (illite, montmorillonite and chlorite). The study was applied to the untreated, filtered and centrifuged fractions of each sample at the pH of the natural waters and at pH 1. The results show that the contribution of dissolved organic matter to the complexing capacity is negligible when compared with SPM. At natural pH, the complexing capacity of filtered and untreated fractions can be described by considering two kinds of binding sites. The associated conditional binding constants are independent of the concentration of suspended matter. Their average logarithms are ca. 6.5 and ca. 4.4. The total concentration of binding sites (S(T)) is in microM range, which is about three orders of magnitude higher than that reported for most of the studied estuaries. This difference is explained on the basis of the great amount of SPM. Hydrodynamic conditions produce variations in the concentration and composition of the SPM. At pH 1 samples still exhibit an important complexing capacity with only one binding site with log K(cond) ca. 5.4. These differences could be attributed to superficial modifications that take place at very low pH.     

Mild hydrolysis of 2-trifluoromethylphenol: kinetics, mechanism and environmental relevance

2-Trifluoromethylphenol was hydrolysed in a phosphate buffer at neutral pH. At mild temperatures ranging from 34 degrees C to 69 degrees C this compound liberates consecutively fluorine anions to form salicylic acid. This process is energetically driven by the hydration of the fluorine anions. No intermediates have been detected by HPLC and (19)F-NMR and this was confirmed by computer calculations which favor the first step in the whole reaction sequence being rate-limiting. Accordingly, the reaction energy of the first dehalogenation of the trifluoromethyl anion is 28.4 kcal mol(-1) higher than for the second dehalogenation. The pseudo-first-order kinetic was determined and from an Arrhenius diagram an activation energy of E(a)=25.1 kcal mol(-1) has been estimated. At 37 degrees C and a pH of 7.4 the half-life was 6.9 h. The rate of hydrolysis was favored at higher pH and it was not influenced by oxygen, sunlight or trace elements found in natural water. The latter was shown by incubations with lake water instead of distilled water.     

Effects of photodegradation of dissolved organic matter on the binding of benzo(a)pyrene

Dissolved organic matter (DOM) in natural waters can bind various organic pollutants, and the affinity of this binding is strongly influenced by the chemical characteristics of the DOM and water pH. This study examined the effects of photochemically induced alteration of the DOM's chemical properties and water pH on the binding of benzo(a)pyrene (BaP). Time- and pH-series of solar-simulated irradiations were performed on a natural water sample and aqueous DOM solutions prepared from aquatic and soil humic substances. The binding affinity of BaP, expressed as a partition coefficient of a compound to DOM, decreased substantially after the DOM samples were irradiated over environmentally relevant radiation doses and pH ranges. The lowering of the pH due to the photoproduction of acidic products often partly offsets the reduction of the binding affinity caused by direct photoalteration of the DOM's chemical structure. The decrease of the binding affinity, after correction for the photoinduced pH change, was positively correlated with the decrease in the molecular weight and the aromaticity of the DOM in the course of irradiation. Increasing O(2) abundance accelerated the decrease of the binding affinity as a result of enhanced DOM photodegradation. Visible light played a more important role in reducing the molecular weight and aromaticity of the DOM than in reducing the content of dissolved organic carbon (DOC) via photoremineralization while the reverse was true for UV radiation, indicating that photochemical reduction of the binding affinity may occur in natural waters at depths greater than UV radiation can reach. A decrease of the affinity of DOM for binding BaP will increase the free dissolved fraction of BaP and thus its availability and toxicity to aquatic organisms. The results from this study may have similar implications for organic pollutants other than BaP.     

电絮凝处理牛仔布印染废水

用铁分别作为电絮凝反应系统的阴极和阳极,研究电絮凝法对牛仔布印染废水的处理效果。考察了电极电压、反应时间和pH等因素对电絮凝法去除实验所用废水中COD和色度效果的影响。结果表明,电极电压和反应时间是主要的影响因素,pH次之。电极电压24 V,反应时间35 min,pH为7.4时,脱色率可达99%,COD去除率在70%左右,处理效果最佳。因此,电絮凝法可以作为印染废水的预处理工艺,有效降低废水COD和色度。     

Investigation of metal ions binding of humic substances using fluorescence emission and synchronous-scan spectroscopy

The binding site interactions of IHSS humic substances, Suwannee River Humic Acid, Suwannee River Fulvic Acid, Nordic Fulvic Acid, and Aldrich Humic Acid with various metals ions and a herbicide, methyl viologen were investigated using fluorescence emission and synchronous-scan spectroscopy. The metal ions used were, Fe(III), Cr(III), Cr(VI), Pb(II), Cu(II) and Ni(II). Stern-Volmer constants, Ksv for these quenchers were determined at pH 4 and 8 using an ionic strength of 0.1 M. For all four humic substances, and at both pH studied, Fe(III) was found to be the most efficient quencher. Quenching efficiency was found to be 3-10 times higher at pH 8. The bimolecular quenching rate constants were found to exceed the maximum considered for diffusion controlled interactions, and indicate that the fluorophore and quencher are in close physical association. Synchronous-scan spectra were found to change with pH and provided useful information on binding site interactions between humic substances and these quenchers.     

The degradation of endocrine disruptor di-n-butyl phthalate by UV irradiation: a photolysis and product study

The direct photolysis of an important endocrine disruptor compound, di-n-butyl phthalate (DBP), has been investigated under monochromatic UV irradiation at 254 nm over a wide pH range (3-11). The investigation was carried out under idealized conditions and has considered both reaction kinetics and the degradation mechanism. It was found that more than 90% of DBP can be degraded within an hour of irradiation in water. A simple model has been developed and used to predict the initial DBP photolysis rate constant at different pH values and initial DBP concentrations. The major decomposition mechanism of DBP is believed to involve the hydrolytic photolysis of the carbon in the alpha and/or beta-position of the ester chain with the production of aromatic carboxylic derivatives. Additionally, multi-degradation pathways are proposed for acid-catalyzed hydrolytic photolysis (pH 3-5), which was found to be useful in explaining the photo-degradation of DBP under acidic conditions. The use of 254 nm UV to photo-degrade DBP was found to be a relatively fast and clean process, especially in neutral to basic conditions.     

Interaction of diuron to human serum albumin: Insights from spectroscopic and molecular docking studies

This investigation was undertaken to determine the interaction of diuron with human serum albumin (HSA) was studied by monitoring the spectral behavior of diuron-HSA system. The fluorescence of HSA at 340 nm excited at 230 nm was obviously quenched by diuron due to dynamic collision and the quenching constant was of the order of 10⁴ L mol^?1 at 310 K. However, no fluorescence quenching was observed when excited at 280 nm. Thermodynamic investigations revealed that the combination between diuron and HSA was entropy driven by predominantly hydrophobic interactions. The binding of diuron induced the drastic reduction in α-helix conformation and the significant enhancement in β-turn conformation of HSA. In addition, both sites marker competition study and molecular modeling simulation evidenced the binding of diuron to HSA primarily took place in subdomain IIIA (Sudlow's site II).     

Copper binding by peat fulvic and humic acids extracted from two horizons of an ombrotrophic peat bog

We studied the binding of Cu(II) to humic acids and fulvic acids extracted from two horizons of an ombrotrophic peat bog by metal titration experiments at pH 4.5, 5.0, 5.5, and 6.0 and 0.1 M KNO3 ionic strength. Free metal ion concentrations in solution were measured using an ion selective electrode. The amounts of base required to maintain constant pH conditions were recorded and used to calculate H+/Cu2+ exchange ratios. The amount of Cu(II) bound to the humic fractions was greater than the amount bound to the fulvic fractions and only at the highest concentrations of metal ion the amount of Cu(II) sorbed by both fractions became equal. The proton to metal ion exchange ratios are similar for all humic substances, with values ranging from 1.0 to 2.0, and decreasing with increased pH. The amount of Cu(II) bound is practically independent of the horizon from which the sample was extracted. The results indicate that the humic substances show similar cation binding behaviour, despite the differences in chemical composition. The copper binding data are quantitatively described with the NICA-Donnan model, which allows to characterize only the carboxylic type binding sites. The values of the binding constants are higher for the humic acids than for the fulvic acids.     

Stability of green fluorescent protein using luminescence spectroscopy: is GFP applicable to field analysis of contaminants?

Green fluorescent protein (GFP) was first isolated in the early 1970s for experimental use from coelenterates or the Pacific jellyfish. Aequorea victoria (Morin and Hastings, 1971). GFP has since become a favored biomarker in the photophysical analysis of molecular and cell biology because of its strong intrinsic visible fluorescence and the feasibility of fusing it to other proteins without affecting their normal functions (Creemers et al., 2000). Here we report using Bacillus subtilis expressing GFP to evaluate the influence of different environmental pH conditions on GFP fluorescence. Emission acquisitions were configured to excite at 471 nm and detect at an emission from 490 to 650 nm at 1-nm increments. Fluorescence intensity was significantly better at pH 7 (4.2 x 105 cps; P-value < 0.01) than at acid or alkaline conditions. GFP is a good biomarker for environments near netural conditions: however, GFP may be unsuitable where soils or waters are below or above pH 7 because of loss in fluorescence intensity. Alternative fluorescent markers and delivery systems must be examined in different environments to optimize responses from bioreporter molecules.     

Role of cupric ions in the H2O2/UV oxidation of humic acids

The purpose of this study is to reveal the role of cupric ions as a natural water contaminant in the H2O2/UV oxidation of humic acids. Humic acids are naturally occurring organic matter and exhibit a strong tendency of complexation with some transition metal ions. Chlorination of humic acids causes potential health hazards due to formation of trihalomethane (THM). The removal of THM precursors has become an issue of public concern. The H2O2/UV process is capable of mineralizing humic acids due to formation of a strong oxidant, hydroxyl radicals, in reaction solution. Experiments were conducted in a re-circulated photoreactor. Different cupric concentrations (0-3.8 mg/l) and different pH values (4-9) were controlled to determine their effects on the degradation of humic acids, UV light absorbance at 254 nm, and H2O2. The presence of cupric ions inhibits humic mineralization and decreases the rate of destruction of humic acids which absorb UV light at 254 nm. On the other hand, the higher the cupric concentration, the lower the H2O2 decomposition rate. In the studied pH range, the minimum of total organic carbon (TOC) removal occurs at pH = 6 in the presence of 2.6 mg/l of cupric ions; both acidification (pH = 4) and alkaline condition (pH = 9) lead to a better removal of TOC. It is inferred from this study that the cupric-complexed form of humic acids is more refractory than the non-complexed one.     

Comparative study of chromium biosorption by red, green and brown seaweed biomass

Dried biomass of the macroalgae Fucus vesiculosus and Fucus spiralis (brown), Ulva spp. (comprising Ulva linza, Ulva compressa and Ulva intestinalis) and Ulva lactuca (green), Palmaria palmata and Polysiphonia lanosa (red) were studied in terms of their chromium biosorption performance. Metal sorption was highly pH dependent with maximum Cr(III) and Cr(VI) sorption occurring at pH 4.5 and pH 2, respectively. Extended equilibrium times were required for Cr(VI) binding over Cr(III) binding (180 and 120min, respectively) thus indicating possible disparities in binding mechanism between chromium oxidation states. The red seaweed P. palmata revealed the highest removal efficiency for both Cr(III) and Cr(VI) at low initial concentrations. However, at high initial metal concentrations F. vesiculosus had the greatest removal efficiency for Cr(III) and performed almost identically to P. lanosa in terms of Cr(VI) removal. The Langmuir Isotherm mathematically described chromium binding to the seaweeds where F. vesiculosus had the largest q(max) for Cr(III) sorption (1.21mmol g(-1)) and P. lanosa had the largest Cr(VI) uptake (0.88mmol g(-1)). P. palmata had the highest affinity for both Cr(III) and Cr(VI) binding with b values of 4.94mM(-1) and 8.64mM(-1), respectively. Fourier transform infrared analysis revealed interactions of amino, carboxyl, sulphonate and hydroxyl groups in chromium binding to Ulva spp. The remaining seaweeds showed involvement of these groups to varying degrees as well as ether group participation in the brown seaweeds and for Cr(VI) binding to the red seaweeds.     

Acid–base physiology response to ocean acidification of two ecologically and economically important holothuroids from contrasting habitats,Holothuria scabra and Holothuria parva

Sea cucumbers are dominant invertebrates in several ecosystems such as coral reefs, seagrass meadows and mangroves. As bioturbators, they have an important ecological role in making available calcium carbonate and nutrients to the rest of the community. However, due to their commercial value, they face overexploitation in the natural environment. On top of that, occurring ocean acidification could impact these organisms, considered sensitive as echinoderms are osmoconformers, high-magnesium calcite producers and have a low metabolism. As a first investigation of the impact of ocean acidification on sea cucumbers, we tested the impact of short-term (6 to 12 days) exposure to ocean acidification (seawater pH 7.7 and 7.4) on two sea cucumbers collected in SW Madagascar, Holothuria scabra, a high commercial value species living in the seagrass meadows, and H. parva, inhabiting the mangroves. The former lives in a habitat with moderate fluctuations of seawater chemistry (driven by day–night differences) while the second lives in a highly variable intertidal environment. In both species, pH of the coelomic fluid was significantly negatively affected by reduced seawater pH, with a pronounced extracellular acidosis in individuals maintained at pH 7.7 and 7.4. This acidosis was due to an increased dissolved inorganic carbon content and pCO₂ of the coelomic fluid, indicating a limited diffusion of the CO₂ towards the external medium. However, respiration and ammonium excretion rates were not affected. No evidence of accumulation of bicarbonate was observed to buffer the coelomic fluid pH. If this acidosis stays uncompensated for when facing long-term exposure, other processes could be affected in both species, eventually leading to impacts on their ecological role.     

微波活化制备加拿大一枝黄花活性炭及对Cd（Ⅱ）的吸附

以入侵植物加拿大一枝黄花为原料,在400℃氮气保护下,直接碳化90 min后,以KOH为活化剂,微波活化的方法制备了高比表面积微孔活性炭SCAC。结果显示活性炭SCAC的表面面积为1 888 m2/g,总孔容量为0.804 cm3/g,微孔容量为0.741 cm3/g,平均孔径0.567 nm,微孔平均孔径0.488 nm。通过静态吸附实验研究了活性炭添加量、溶液初始浓度、初始pH及吸附时间对SCAC吸附Cd(Ⅱ)的影响,通过动力学方程拟合探讨了活性炭对Cd(Ⅱ)的吸附机理。结果表明,吸附平衡时间、最佳pH及活性炭添加量分别为120 min、pH>7.5及0.05 g/50 mL,活性炭SCAC吸附动力学过程符合准二级方程模型。