Removal of dinitrotoluenes from water via reduction with iron and peroxidase-catalyzed oxidative polymerization: a comparison between Arthromyces ramosus peroxidase and soybean peroxidase

A two-step process for the removal of dinitrotoluene from water is presented: zero-valent iron reduction is coupled with peroxidase-catalyzed polymerization of the resulting diaminotoluenes (DAT). The effect of pH was examined in the reduction step: at pH 6 the reaction occurred much more rapidly than at pH 8. In the second step, optimal pH and substrate ratio, minimal enzyme concentration and effect of polyethylene glycol (PEG) as an additive for greater than 95% conversion of DAT, over a 3h reaction period were determined using high performance liquid chromatography. Two enzymes were investigated and compared: Arthromyces ramosus peroxidase (ARP) and soybean peroxidase (SBP). The optimal pH values were 5.4 and 5.2 for ARP and SBP, respectively, but SBP was more resistant to mild acid whereas ARP was more stable in neutral solutions. SBP was found to have a greater hydrogen peroxide demand (optimal peroxide/DAT molar ratio for SBP: 2.0 and 3.0 for 2,4-diaminotoluene (2,4-DAT) and 2,6-diaminotoluene (2,6-DAT), respectively; for ARP: 1.5 and 2.75 for 2,4-DAT and 2,6-DAT, respectively) but required significantly less enzyme (0.01 and 0.1 U ml(-1) for 2,4-DAT and 2,6-DAT, respectively) to convert the DAT than ARP (0.4 and 1.5 U ml(-1) for 2,4-DAT and 2,6-DAT, respectively). PEG was shown to have no effect upon the degree of substrate conversion for either enzyme.     

Comparison of the removal of 2,4-dichlorophenol and phenol from polluted water, by peroxidases from tomato hairy roots, and protective effect of polyethylene glycol

Multiple efforts have been directed towards optimized processes in which enzymes, like peroxidases, are used to remove phenolic compounds from polluted wastewater. Here we describe the use of peroxidase isoenzymes from tomato hairy roots, which were able to oxidise 2,4-dichlorophenol (2,4-DCP) and phenol from aqueous solutions. This could be an interesting alternative for the removal of these compounds from contaminated sites. We used different enzyme fractions: total peroxidases (TP), ionically bound to cell wall peroxidases (IBP), basic (BP) and acidic peroxidases (AP). We analyzed the optimum conditions of removal, the effect of Polyethyleneglycol (PEG-3350) on the process and on the enzyme activities, to obtain the maximum efficiency. The optimal H2O2 concentrations for 2,4-DCP and phenol removal were 1 and 0.1mM, respectively. TP, IBP and BP showed better removal efficiencies than AP, for both contaminants. The addition of different concentrations (10-100mg l(-1)) of PEG-3350 to solutions containing 2,4-DCP showed no effect on the removal efficiencies of the isoenzymes. However, PEG (100mg l(-1)) increased the removal efficiency of phenol by BP and IBP fractions. On the other hand, peroxidase activities from BP and IBP fractions were 3 and 13 times higher, respectively, than those detected for the same fractions in phenol treated solutions without PEG. The protective effect of PEG, which depends on the contaminant as well as of the enzyme fraction used, would be important to improve the removal efficiency of phenol by some peroxidase isoenzymes.     

凹凸棒石粘土固定辣根过氧化物酶处理含酚废水

采用凹凸棒石粘土、可溶性淀粉和工业水玻璃作为制备凹凸棒石粘土基颗粒的材料,并将制备的凹凸棒石粘土基颗粒进行改性,得到了改性后的凹凸棒石基多孔材料,并将其作为固定辣根过氧化物酶的载体,再应用于含酚废水的处理研究,取得了良好的效果。实验结果表明,辣根过氧化物酶的最佳固定化条件为:单位酶活载体量1 mg、固定化时间1.5小时、固定pH值5。并且在固定化酶循环使用6次后,苯酚去除率仍能达到62.3%。     

Application of Brassica napus hairy root cultures for phenol removal from aqueous solutions

Phenolic compounds present in the drainage from several industries are harmful pollutants and represent a potential danger to human health. In this work we have studied the removal of phenol from water using Brassica napus hairy roots as a source of enzymes, such as peroxidases, which were able to oxidise phenol. These hairy roots were investigated for their tolerance to highly toxic concentrations of phenol and for the involvement of their peroxidase isoenzymes in the removal of phenol. Roots grew normally in medium containing phenol in concentrations not exceeding 100 mg l(-1), without the addition of H(2)O(2). However, roots were able to remove phenol concentrations up to 500 mg l(-1), in the presence of H(2)O(2), reaching high removal efficiency, within 1h of treatment and over a wide range of pH (4-9). Hairy roots could be re-used, at least, for three to four consecutive cycles. Peroxidase activity gradually decreased to approximately 20% of the control, at the fifth cycle. Basic and near neutral isoenzymes (BNP) decreased along time of recycling while acidic isoenzymes (AP) remained without changes. Although both group of isoenzymes would be involved in phenol removal, AP showed higher affinity and catalytic efficiency for phenol as substrate than BNP. In addition, AP retained more activity than BNP after phenol treatment. Thus, AP appears to be a promising isoenzyme for phenol removal and for application in continuous treatments. Furthermore, enzyme isolation might not be necessary and the entire hairy roots, might constitute less expensive enzymatic systems for decontamination processes.     

Treatment of phenolic wastewater by horseradish peroxidase immobilized by bioaffinity layering

Horseradish peroxidase was immobilized by bioaffinity layering and used for the treatment of wastewater containing p-chlorophenol. For this purpose, lectin Concanavalin A was bound to Sephadex beads. The glycoenzyme peroxidase was layered upon this Con A layer. Subsequently, alternate layers of the enzyme and Con A were applied. The most efficient design consisted of three layers of Con A and peroxidase each. This immobilized enzyme preparation retained 80% of the activity of the free peroxidase used for immobilization. PEG at the concentration of 0.1 mg ml(-1) was found to prevent enzyme inactivation by the products, although it increased the process time. Thus 60 U ml(-1) of enzyme completely converted the p-chlorophenol (into products) in 4 min in the absence of PEG. On the other hand, only 0.05 U ml(-1) of enzyme was required for this purpose in the presence of PEG but the process required 60 min. Peroxidase converts phenol molecules into free radicals. These free radicals then polymerize and get precipitated. As a further means of minimizing exposure of the enzyme to free radicals and enhancing the reusability, it was decided to remove the enzyme from reaction medium after 10 min. With this strategy, the bioaffinity layered peroxidase preparation could be reused five times without any loss of activity.     

Acid azo dye degradation by free and immobilized horseradish peroxidase (HRP) catalyzed process

Acid azo (Acid Black 10 BX) dye removal by plant based peroxidase catalyzed reaction was investigated. Horseradish peroxidase (HRP) was extracted from horseradish roots and its performance was evaluated in both free and immobilized form. HRP showed its ability to degrade the dye in aqueous phase. Studies are further carried out to understand the process parameters such as aqueous phase pH, H2O2 dose, dye and enzyme concentrations during enzyme-mediated dye degradation process. Experimental data revealed that dye (substrate) concentration, aqueous phase pH, enzyme and H2O2 dose play a significant role on the overall enzyme-mediated reaction. Acrylamide gel immobilized HRP showed effective performance compared to free HRP and alginate entrapped HRP. Alginate entrapped HRP showed inferior performance over the free enzyme due to the consequence of non-availability of the enzyme to the dye molecule due to polymeric immobilization. Standard plating studies performed with Pseudomonas putida showed enhanced degradation of HRP catalyzed dye compared to control.     

Peroxidase-mediated removal of endocrine disrupting compound mixtures from water

Several classes of oxidative enzymes have shown promise for efficient removal of endocrine disrupting compounds (EDCs) that are resistant to conventional wastewater treatments. Although the kinetics of reactions between individual EDCs and selected oxidative enzymes are well documented in the literature, there has been little investigation of reactions with EDC mixtures. This makes it impossible to predict how enzyme-mediated treatment systems will perform since wastewater effluents generally contain multiple EDCs. This paper reports pseudo-first order rate constants for a model oxidative enzyme, horseradish peroxidase (HRP), during single-substrate (k₁) and mixed-substrate (k_1-MIX) reactions. Measured values are compared with literature values of three Michaelis-Menten parameters: half-saturation constant (K_M), enzyme turnover number (k_CAT), and the ratio k_CAT/K_M. Published reports had suggested that each of these could be correlated with HRP reactivity towards EDCs in mixtures, and empirical results from this study show that K_M can be used to predict the sequence of EDC removal reactions within a particular mixture. We also observed that k_1-MIX values were generally greater than k₁ values and that compounds exhibiting greatest estrogenic toxicities reacted most rapidly in a given mixture. Finally, because K_M may be tedious to measure for every EDC of interest, we have constructed a quantitative structure-activity relationship (QSAR) model to predict these values. This model predicts K_M quite accurately (R² = 89%) based on two molecular characteristics: molecular volume and hydration energy. Its accuracy makes this QSAR a useful tool for predicting which EDCs will be removed most efficiently during enzyme treatment of EDC mixtures.     

Coupled oxidation of aromatic hydrocarbons by horseradish peroxidase and hydrogen peroxide

The oxidation capability of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2) coupled oxidation of aromatic hydrocarbons (o-xylene-d10 and naphthalene-d8) was investigated. Batch experiments were conducted using horseradish peroxidase prepared in potassium phosphate buffer in the presence of H2O2. The oxidation of aromatic hydrocarbon was tested as a function of HRP at a fixed concentration of H2O2, and as a function of the concentration of H2O2 at a constant HRP activity (4000 units/ml). The mass removal of o-xylene-d10 and naphthalene-ds increased with increasing HRP enzymatic activity, and up to 54% and 51% of mass removal were observed for o-xylene-d10 and naphthalene-d8, respectively. Increasing the concentration of H2O2 resulted in increased mass removal of aromatic hydrocarbons.     

Phytoremediation of phenol using Vicia sativa L. plants and its antioxidative response

Common vetch (Vicia sativa L.) is a legume species with an extensive agricultural use. However, the phytoremediation potentiality of this species has not been sufficiently explored because little is known about its resistance to inorganic and organic pollutants. In the present work, phenol tolerance of common vetch was assayed at different stages of growth. Germination index and germination rate decreased only at high phenol concentrations (250 and 500 mg L(-1)), whereas 30-day-old plants were able to tolerate this pollutant, with high removal efficiencies. The activities of antioxidative enzymes, such as peroxidase (POD) and ascorbate peroxidase, increased significantly with the highest phenol concentration, whereas superoxide dismutase activity, malondialdehyde, and H(2)O(2) levels remained unaltered. Besides, an increase in two basic isoforms of POD was observed in plants treated with phenol. The results suggested that common vetch has an efficient protection mechanism against phenol-induced oxidative damage. Moreover, it could tolerate and remove high phenol concentrations, avoiding serious phytotoxic effects. Thus, V. sativa could be considered an interesting tool in the field of phytoremediation.     

Potential applications of immobilized bitter gourd (Momordica charantia) peroxidase in the removal of phenols from polluted water

The potential applications of immobilized bitter gourd peroxidase in the treatment of model wastewater contaminated with phenols have been investigated. The synthetic water was treated with soluble and immobilized enzyme preparations under various experimental conditions. Maximum removal of phenols was found in the buffers of pH values 5.0-6.0 and at 40 degrees C in the presence of 0.75 mM H(2)O(2). Fourteen different phenols were independently treated with soluble and immobilized bitter gourd peroxidase in the buffer of pH 5.6 at 37 degrees C. Chlorinated phenols and native phenol were significantly removed while other substituted phenols were marginally removed by the treatment. Phloroglucinol and pyrogallol were recalcitrant to the action of bitter gourd peroxidase. Immobilized bitter gourd peroxidase preparation was capable of removing remarkably high percentage of phenols from the phenolic mixtures. Significantly higher level of total organic carbon was removed from the model wastewater containing individual phenol or complex mixture of phenols by immobilized bitter gourd peroxidase as compared to the soluble enzyme. 2,4-dichlorophenol and a phenolic mixture were also treated in a stirred batch reactor with fixed quantity of enzyme for longer duration. The soluble bitter gourd peroxidase ceased to function after 3h while the immobilized enzyme was active even after 6h of incubation with phenolic solutions.     

Fe (III) supported on resin as effective catalyst for the heterogeneous oxidation of phenol in aqueous solution

FeIII supported on resin as an effective catalyst for oxidation was prepared and applied for the degradation of aqueous phenol. Phenol was selected as a model pollutant and the catalytic oxidation was carried out in a batch reactor using hydrogen peroxide as the oxidant. The influent factors on oxidation, such as catalyst dosage, H2O2 concentration, pH, and phenol concentration were examined by considering both phenol conversion and chemical oxygen demand (COD) removal. The FeIII-resin catalyst possesses a high oxidation activity for phenol degradation in aqueous solution. The experimental results of this study show that almost 100% phenol conversion and over 80% COD removal can be achieved with the FeIII-resin catalyst catalytic oxidation system. A series of prepared resin were investigated for improving the oxidation efficiency. It was found that the reaction temperature and initial pH in solution significantly affected both of phenol conversion and COD removal efficiency. The activity of the catalyst significantly decreased at high pH, which was similar to the Fenton-like reaction mechanism. Results in this study indicate that the FeIII-resin catalytic oxidation process is an efficient method for the treatment of phenolic wastewater.     

Comparison of lignin peroxidase and horseradish peroxidase for catalyzing the removal of nonylphenol from water

Concentrations of aqueous-phase nonylphenol (NP), a well-known endocrine-disrupting chemical, are shown to be reduced effectively via reaction with lignin peroxidase (LiP) or horseradish peroxidase (HRP) and hydrogen peroxide. We systematically assessed their reaction efficiencies at varying conditions, and the results have confirmed that the catalytic performance of LiP toward NP was more efficient than that of HRP under experimental conditions. Mass spectrum analysis demonstrated that polymerization through radical–radical coupling mechanism was the pathway leading to NP transformation. Our molecular modeling with the assistance of ab initio suggested the coupling of NP likely proceeded via covalent bonding between two NP radicals at their unsubstituted carbons in phenolic rings. Data from acute immobilization tests with Daphnia confirm that NP toxicity is effectively eliminated by LiP/HRP-catalyzed NP removal. The findings in this study provide useful information for understanding LiP/HRP-mediated NP reactions, and comparison of enzymatic performance can present their advantages for up-scale applications in water/wastewater treatment.     

高铁酸盐氧化降解水中苯酚的动力学及机理研究

以自制高铁酸钾(K₂FeO₄)为原料,探讨了影响高铁酸盐氧化降解苯酚的主要因素,并研究了苯酚降解的动力学特征和反应机理。结果表明,高铁酸盐加入量、pH值、持续搅拌、反应温度、反应时间都影响苯酚去除效果。其中高铁酸盐加入量是影响苯酚去除效果的关键因素,高铁酸盐氧化降解苯酚的最佳pH值范围为9～10,持续搅拌和提高反应温度只影响苯酚降解速率而不影响苯酚降解率。苯酚的降解过程遵循一级反应动力学模型。苯酚被高铁酸盐氧化生成CO₂、H₂O以及一部分难矿化的有机物。     

The treatment of waste air containing phenol vapors in biotrickling filter

This research aimed at investigating the biodegradation of phenol contaminated-air streams in biotrickling filter. The effect of inlet concentration (200-1000 ppmv) and empty bed contact time (EBCT) (15-60 s) were investigated under steady state, transient and shock loading, and shutdown periods. Upon rapid start up operation, inlet phenol concentrations of up to 1000 ppmv did not significantly affect the performance of the biotrickling filter at EBCT of 60 s, so that removal efficiency was well greater than 99%. In addition, the EBCT as low as 30 s did not have detrimental effects on the efficiency of the bioreactor and phenol removal was greater than 99%. Decreasing the EBCT to 15s reduced the removal efficiency to around 92%. The maximum elimination capacity obtained in the biotrickling filter was 642 g(phenol) m(-3) h(-1), where the removal efficiency was only 57%. Results from the transient loading experiments revealed that the biotrickling filter could effectively handle the variations of the inlet loads without the phenol removal capacity being significantly affected.     

Application of pier waste sludge for catalytic activation of proxy-monosulfate and phenol elimination from a petrochemical wastewater

This investigation aimed to remove phenol from real wastewater (taken from a petrochemical company) by activating peroxy-monosulfate (PMS) using catalysts extracted from pier waste sludge. The physical and chemical properties of the catalyst were evaluated by FE-SEM/EDS, XRD, FTIR, and TGA/DTG tests. The functional groups of O–H, C–H, CO₃^2?, C–H, C–O, N–H, and C–N were identified on the catalyst surface. Also, the crystallinity of the catalyst before and after reaction with petrochemical wastewater was 103.4 nm and 55.8 nm, respectively. Operational parameters of pH (3–9), catalyst dose (0–100 mg/L), phenol concentration (50–250 mg/L), and PMS concentration (0–250 mg/L) were tested to remove phenol. The highest phenol removal rate (94%) was obtained at pH=3, catalyst dose of 80 mg/L, phenol concentration of 50 mg/L, PMS concentration of 150 mg/L, and contact time of 150 min. Phenol decomposition in petrochemical wastewater followed the first-order kinetics (k> 0.008 min^?1, R²> 0.94). Changes in pH factor were very effective on phenol removal efficiency, and maximum efficiency (≈83%) was achieved in pH 3. The catalyst stability test was performed for up to five cycles, and phenol removal in the fifth cycle was reduced to 42%. Also, the energy consumption in this study was 77.69 kW h/m³. According to the results, the pier waste sludge catalyst/PMS system is a critical process for eliminating phenol from petrochemical wastewater.

     

吸附-氧化法应急处理饮用水源突发苯酚污染的研究

以黄浦江上游水源地突发苯酚污染为背景,重点考察了粉末活性炭（PAC）吸附、高锰酸钾（KMnO4）氧化及两者联用技术的除酚效能。结果表明,活性炭及氧化剂种类的选择是影响处理效果的重要因素,微孔发达、比表面积巨大的竹炭对苯酚的去除效果明显优于煤质炭、椰壳炭和木质炭;KMnO4对苯酚的氧化能力强于次氯酸钠和高铁酸钾。增大PAC和KMnO4的投加量,可有效提高对苯酚的去除率;PAC吸附-KMnO4氧化联用技术可大大提高除酚效能,投加50mg／LPAC,2mg／LKMnO4可将初始浓度为250／μg／L和500／μg／L的含酚原水分别处理至18μg／L和66／μg／L,是应对高浓度苯酚突发污染的有效应急措施。     

Biodegradation of 2,4,6-trichlorophenol in the presence of primary substrate by immobilized pure culture bacteria

In this study, pure strains that are capable of utilizing 2,4,6-trichlorophenol have been isolated from the mixed culture grown on substrates containing chlorophenolic compounds. Studies have been carried out on the capability of these isolated pure strains in suspended and immobilized forms to decompose 2,4,6-trichlorophenol. Additionally, the influence of primary substrates (e.g., phenol, 2-chlorophenol, 3-chlorophenol, 4-chlorophenol, 2,4-dichlorophenol) on the decomposition of 2,4,6-trichlorophenol by the isolated pure strains grown in immobilized form is also investigated. The results are: Through bacterial isolation and identification, three pure strains have been obtained: Pseudomonas spp. strain 01, Pseudomonas spp. strain 02 and Agrobacterium spp. Whether in suspended or immobilized forms, all strains have poor removal efficiencies of 2,4,6-trichlorophenol. However, addition of 200 mg/l phenol will enable the immobilized Pseudomonas spp. strain 01, and Pseudomonas spp. strain 02 to achieve 65% and 48% removal of 2,4,6-trichlorophenol, respectively. Addition of phenol will assist the immobilized Pseudomonas spp. strain 02 in achieving removal of 2,4,6-trichlorophenol but the removal efficiency is not good if the phenol concentration is too low. The optimum phenol concentration should be between 200 and 400 mg/l.     

A combination of electrokinetics and Pd/Fe PRB for the remediation of pentachlorophenol-contaminated soil

Electrokinetic (EK) remediation of pentachlorophenol (PCP)-contaminated soil is difficult because PCP dissociates at different pH values along soil column and shows different transport behaviors near anode and cathode. In the present study, a permeable reactive barrier (PRB) filled with reactive Pd/Fe particles was installed between anode and cathode to reach the dechlorination of PCP during its EK movement. When PRB was installed at the position of 0.3 (normalized distance from anode), PCP in the section from anode to PRB could transport through PRB, while PCP in the section from cathode to PRB was accumulated near PRB. PCP was hardly dechlorinated by PRB wherein high pH was reached. When PRB was installed at the position of 0.5 and the pH in the PRB was decreased by periodical injection of HAc, 49% of PCP was removed, and 22.9% was recovered as phenol which was mostly collected in catholyte. The mechanism of PCP removal was proposed as the EK movement of PCP into the PRB compartment, the complete dechlorination of PCP to phenol by Pd/Fe in the PRB compartment, and the subsequent removal of phenol by electroosmosis. This study proved that the combination of electrokinetics and Pd/Fe PRB was effective for the remediation of PCP-contaminated soil.     

Removal of phenol using mushrooms and immobilized polyphenol oxidase in a plug flow reactor

Phenols are toxic and hazardous pollutants, hence their removal from industrial effluents is important. In this study, edible mushroom particles, which are a cheap and natural source of polyphenol oxidase (PPO), were used directly for phenol removal in a plug flow reactor. Initially, 60% phenol removal was achieved, but this percentage decreased after a second use. When the reactor included PPO immobilised in chitosan gels, the percentage of phenol removal was 30%, and was stable even after the twelfth use.     

Phenol removal from wastewater by adsorption on zeolitic composite

It is well known that adsorption is an efficient method of removal of various pollutants from wastewater. The present study examines the phenol removal from water by adsorption on a new material, based on zeolitic volcanic tuff. This compound contains zeolitic tuff and cellulose, another known adsorbent, in a mass ratio of 4 to 1. The performances of the new adsorbent composite were compared with those of a widely used adsorbent material, zeolitic volcanic tuff. The adsorbent properties were tested on batch synthetic solutions containing 1–10 mg L^?1 (1–10 ppm) phenol, at room temperature without pH adjustment. The influence of the adsorbent dose, pH and contact time on the removal degree of phenol from water was investigated. The experimental data were modeled using the Langmuir, Freundlich, and Temkin adsorption isotherms. The Langmuir model was found to best represent our data revealing a monolayer adsorption with a maximum adsorption capacity between 0.12 and 0.53 mg g^?1 at 25 °C, for 2.00 g of adsorbent, depending on the initial phenol concentration. The adsorption kinetic study was performed using a pseudo-first- and pseudo-second-order kinetic models illustrating that phenol adsorption on zeolite composite is well described by pseudo-first kinetic equations. Our results indicated that phenol adsorption on the new adsorbent composite is superior to that on the classic zeolite.