Exploring the potential of biofiltration for mitigating harmful gaseous emissions from small or old landfills: a review.

Landfills are widely employed as the primary means of solid waste disposal. However, this practice generates landfill gas (LFG) which contains methane (CH4), a potent greenhouse gas, as well as various volatile organic compounds and volatile inorganic compounds. These emissions from landfills contribute to approximately 25% of the total atmospheric CH4, indicating the imperative need to valorize or treat LFG prior to its release into the atmosphere. This review first aims to outline landfills, waste disposal and valorization, conventional gas treatment techniques commonly employed for LFG treatment, such as flares and thermal oxidation. Furthermore, it explores biotechnological approaches as more technically and economically feasible alternatives for mitigating LFG emissions, especially in the case of small and aged landfills where CH4 concentrations are often below 3% v/v. Finally, this review highlights biofilters as the most suitable biotechnological solution for LFG treatment and discusses several advantages and challenges associated with their implementation in the landfill environment.

Effects of potential inducers to enhance laccase production and evaluating concomitant enzyme immobilisation.

This work investigated non-polar solvent hexane and polar solvents methanol and ethanol as inducers besides a well-known inducer, copper, for laccase production with and without mesoporous silica-covered plastic packing under sterilised and unsterilised conditions. The potential of waste-hexane water, which is generated during the mesoporous silica production process, was also investigated as a laccase inducer. During the study, the free and immobilised laccase activity on the packing was measured. The results showed that the highest total laccase activity, approximately 10,000 Units, was obtained under sterilised conditions with 0.5 mM copper concentration. However, no immobilised laccase activity was detected except in the copper and ethanol sets under unsterilised conditions. The maximum immobilised laccase activity of the sets that used waste hexane as an inducer was 1.25 U/mg packing. According to its significant performance, waste hexane can be an alternative inducer under sterilised conditions. Concomitant immobilised packing showed satisfactory laccase activities and could be a promising method to reduce operation costs and improve the cost-efficiency of enzymatic processes in wastewater treatment plants.

Prolonged operation of a methane biofilter from acclimation to the failure stage.

Global warming needs immediate attention to reduce major greenhouse gas emissions such as methane (CH4). Bio-oxidation of dilute CH4 emissions in packed-bed bioreactors such as biofilters has been carried out over recent years at laboratory and large scales. However, a big challenge is to keep CH4 biofilters running for a long period. In this study, a packed-bed lab-scale bioreactor with a specialized inorganic-based filter bed was successfully operated over four years for CH4 elimination. The inoculation of the bioreactor was the active leachate of another CH4 biofilter which resulted in a fast acclimation and removal efficiency (RE) reached 80% after seven weeks of operation for CH4 inlet concentrations ranging from 700 to 800 ppmv and an empty bed residence time (EBRT) of 6 min. During four years of operation, the bioreactor often recorded REs higher than 65% for inlet concentrations in the range of 1900-2200 ppmv and an EBRT of 6 min. The rate and interval of the nutrient supply played an important role in maintaining the bioreactor's high performance over the long operation. Forced shutdowns were unavoidable during the 4-year operation and the bioreactor fully tolerated them with a partial recovery within one week and a progressive recovery over time. In the end, the bioreactor's filter bed started to deteriorate due to a long shutdown of twelve weeks and the extended operation of four years when the RE dropped to below 8% with no sign of returning to its earlier performance.

Air biofilters for a mixture of organic gaseous pollutants: an approach for industrial applications.

Hazardous airborne pollutants are frequently emitted to the atmosphere in the form of a gaseous mixture. Air biofilters as the primary biotechnological choice for waste gas treatment (low inlet concentration and high gas flow rate) should run properly when the feed contains multiple pollutants. Simultaneous removal of pollutants in biofilters has been extensively studied over the last 10 years. In this review, the results and findings of the mentioned studies including different groups of pollutants, such as methane (CH4) and volatile organic compounds (VOCs) are discussed. As the number of pollutants in a mixture increases, their elimination might become more complicated due to interactions between the pollutants. Parallel batch studies might be helpful to better understand these interaction effects in the absence of mass transfer limitations. Setting optimum operating conditions for removal of mixtures in biofilters is challenging because of opposing properties of pollutants. In biofilters, concerns, such as inlet gas composition variation and stability while dealing with abrupt inlet load and concentration changes, must be managed especially at industrial scales. Biofilters designed with multi-layer beds, allow tracking the fate of each pollutant as well as analyzing the diversity of microbial culture across the filter bed. Certain strategies are recommended to improve the performance of biofilters treating mixtures. For example, addition of (bio)surfactants as well as a second liquid phase in biotrickling filters might be considered for the elimination of multiple pollutants especially when hydrophobic pollutants are involved.

Bioelimination of low methane concentrations emitted from wastewater treatment plants: a review.

Sewage from residents and industries is collected and transported to wastewater treatment plants (WWTPs) with sewer networks. The operation of WWTPs results in emissions of greenhouse gases, such as methane (CH4), mostly due to sludge anaerobic digestion. Amounts of emissions depend on the source of influent, i.e. municipal and industrial wastewater as well as sewer systems (gravity and rising). Wastewater is the fifth-largest source of anthropogenic CH4 emissions in the world and represents 7-9% of total global CH4 emissions into the atmosphere. Global wastewater CH4 emission grew by approximately 20% from 2005 to 2020 and is expected to grow by 8% between 2020 and 2030, which makes wastewater an important CH4 emitter worldwide. This review initially considers the emission of CH4 from WWTPs and sewer networks. In the second part, biotechniques available for biodegradation of low CH4 concentrations (<5% v/v) encountered in WWTPs have been studied. The paper reviews major bioreactor configurations for the treatment of polluted air, i.e. biotrickling filters, bioscrubbers, two-liquid phase bioreactors, biofilters, and hybrid reactor configurations, after which it focuses on CH4 biofiltration systems. Biofiltration represents a simple and efficient approach to bio-oxidize CH4 in waste gases from WWTPs. Major factors influencing a biofilter's performance along with knowledge gaps in relation to its application for treating gaseous emissions from WWTPs are discussed.

Simultaneous biodegradation of methane and styrene in biofilters packed with inorganic supports: Experimental and macrokinetic study.

Four upflow 0.018 m3 biofilters (3 beds), B-ME, B-200, B-500 and B-700, all packed with inorganic materials, were operated at a constant air flow rate of 0.18 m3 h-1 to eliminate methane (CH4), a harmful greenhouse gas (GHG), and styrene (C8H8), a carcinogenic volatile organic compound (VOC). The biofilters were irrigated with 0.001 m3 of recycled nutrient solution (NS) every day (flow rate of 60 × 10-3 m3 h-1). Styrene inlet load (IL) was kept constant in each biofilter. Different CH4-ILs varying in the range of 7-60 gCH4 m-3 h-1 were examined in B-ME (IL of 0 gC8H8 m-3 h-1), B-200 (IL of 9 gC8H8 m-3 h-1), B-500 (IL of 22 gC8H8 m-3 h-1) and B-700 (IL of 32 gC8H8 m-3 h-1). Finally, the effect of C8H8 on the macrokinetic parameters of CH4 biofiltration was studied based on the Michaelis-Menten model. Average C8H8 removal efficiencies (RE) varying between 64 and 100% were obtained at CH4-ILs increasing from 7 to 60 gCH4 m-3 h-1 and for C8H8-ILs range of 0-32 gC8H8 m-3 h-1. More than 90% of C8H8 was removed in the bottom and middle beds of the biofilters. By increasing C8H8-IL from 0 to 32 gC8H8 m-3 h-1, maximal EC in Michaelis-Menten model and macrokinetic saturation constant declined from 311 to 39 g m-3 h-1 and from 19 to 2.3 g m-3, respectively, which confirmed that an uncompetitive inhibition occurred during CH4 biofiltration in the presence of C8H8.

A hybrid bioreactor based on insolubilized tyrosinase and laccase catalysis and microfiltration membrane remove pharmaceuticals from wastewater.

The increasing presence of pharmaceutical products (PPs) and other organic contaminants of emerging concern (CECs) in aquatic systems has become one of the major global environmental contamination concerns. Sewage treatment plants (STPs) are one of the major sources of PPs discharge into natural waters due to the deficiencies of conventional treatment processes to deal with these micropollutants. Numerous new treatment processes and technologies have been investigated for the removals of CECs in wastewaters with more or less success. In the present study, we investigated the efficiency of a hybrid bioreactor (HBR) of a combined crosslinked tyrosinase and laccase aggregates and hollow fiber microfiltration (MF) membrane to remove a mixture of 14 PPs from municipal wastewater at environmentally relevant concentration of 10 µg/L. After a 5-day continuous operation, the HBR achieved complete removal of all tested PPs. Results also highlight that these high performances result from a synergistic action of the MF membrane and the insoluble enzymes. The biocatalyst retained nearly 70% of its initial enzymatic activity over the treatment period. The removal of PPs is unlikely to result from their sole sorption on the membrane. Overall, the results suggest that the HBR is well suited to the biocatalysts (i.e. insolubilized tyrosinase and laccase). The results invite to further investigate how the HBR can be tailored with various types of enzymes and membranes for either specific or non-specific target substrates and to further explore the applicability of this technology for the continuous treatment of wastewater at environmentally relevant concentration of PPs.

Steady state and dynamic behaviors of a methane biofilter under periodic addition of ethanol vapors.

Ethanol was added to a methane (CH4) biofilter with inorganic packing materials over three cycles based on increasing the gas flow rates from 3 to 6 and finally to 12 L min-1 corresponding to empty bed residence times (EBRT) of 6, 3 and 1.5 min. The steady state performance of the CH4 biofilter was studied for CH4 inlet loads (ILs) of 33, 66 and 132 gCH4 m-3 h-1 prior and after each ethanol cycle. In addition, the steady state removal of a mixture of CH4 and ethanol for a CH4/ethanol mass ratio of around 7.5 gCH4 g -1ethanol was evaluated over three cycles (EBRTs of 6, 3 and 1.5 min). In the absence of ethanol, the CH4 removal efficiency (RE) dropped from 35 to 7% due to an EBRT decrease from 6 to 1.5 min. In addition, the presence of ethanol resulted in a CH4 RE reduction at a constant EBRT in every cycle. The CH4 REs dropped from 35 to 29%, 17 to 13% and 7 to 0% for corresponding ethanol ILs of 4.5, 9 and 18 gethanol m-3 h-1 over the cycles. Moreover, the periodic presence of ethanol in the CH4 biofilter allowed the study of transient behaviors of the biofilter during ethanol addition and the biofilter recovery after each cycle. The CH4 RE reduction as a result of ethanol addition in each cycle was instantaneous. However, the CH4 RE recovery after completion of ethanol addition took 10, 14 and 25 days for ethanol ILs of 4.5, 9 and 18 gethanol m-3 h-1 respectively. The recovery time was related to the ethanol concentration in the leachate which were 1100 ± 200, 1100 ± 350 and 2500 ± 400 gethanol m-3leachate for corresponding ethanol ILs of 4.5, 9 and 18 gethanol m-3 h-1, respectively. Based on steady state and dynamic process conditions of the biofilter, the lowest gas flow rate of 3 L min-1 (EBRT of 6 min) produced the best performance when both pollutants were present (CH4 IL of 33 gCH4 m-3 h-1 and ethanol IL of 4.5 gethanol m-3 h-1).

Hybrid bioreactor (HBR) of hollow fiber microfilter membrane and cross-linked laccase aggregates eliminate aromatic pharmaceuticals in wastewaters.

Widespread detection of numerous micropollutants including aromatic pharmaceuticals in effluents of wastewater treatment plants has prompted much research aimed at efficiently eliminating these contaminants of environmental concerns. In the present work, a novel hybrid bioreactor (HBR) of cross-linked enzymes aggregates of laccase (CLEA-Lac) and polysulfone hollow fiber MF membrane was developed for the elimination of acetaminophen (ACT), mefenamic acid (MFA) and carbamazepine (CBZ) as model aromatic pharmaceuticals. The MF alone showed removals of the three drugs varying approximately from 50 to 90% over the course of 8h in the filtrate of aqueous solution. Synergistic action of the MF and CLEA-Lac during operation achieved eliminations from aqueous solution of around 99%, nearly 100% and up to 85% for ACT, MFA and CBZ, respectively. Under continuous operation, the HBR demonstrated elimination rates of the drugs from filtered wastewater up to 93% after 72h for CBZ and near complete elimination of ACT and MFA was achieved within 24h of treatment. Concomitantly to the drugs eliminations in the wastewater, the CLEA-Lac exhibited 25% residual activity while being continuously recycled with no activity in the filtrate. Meanwhile, the filtrate flowrate showed only minor decline indicating limited fouling of the membrane.

Synthesis and characterization of combined cross-linked laccase and tyrosinase aggregates transforming acetaminophen as a model phenolic compound in wastewaters.

Laccase (EC 1.10.3.2) and tyrosinases (EC 1.14.18.1) are ubiquitous enzymes present in nature as they are known to originate from bacteria, fungi, plants, etc. Both laccase and tyrosinase are copper-containing phenoloxidases requiring readily available O2 without auxiliary cofactor for their catalytic transformation of numerous phenolic substrates. In the present study, laccase and tyrosinase have been insolubilized as combined crosslinked enzyme aggregates (combi-CLEA) using chitosan, a renewable and biodegradable polymer, as crosslinker. The combi-CLEA, with specific activity of 12.3 U/g for laccase and 167.4 U/g for tyrosinase, exhibited high enzymatic activity at pH5-8 and temperature at 5-30°C, significant resistance to denaturation and no diffusional restriction to its active site based upon the Michaelis-Menten kinetic parameters. Subsequently, the combi-CLEA was applied to the transformation of acetaminophen as a model phenolic compound in samples of real wastewaters in order to evaluate the potential efficiency of the biocatalyst. In batch mode the combi-CLEA transformed more than 80% to nearly 100% of acetaminophen from the municipal wastewater and more than 90% from the hospital wastewater. UPLC-MS analysis of acetaminophen metabolites showed the formation of its oligomers as dimers, trimers and tetramers due to the laccase and 3-hydroxyacetaminophen due to the tyrosinase.

Laccase immobilization and insolubilization: from fundamentals to applications for the elimination of emerging contaminants in wastewater treatment.

Over the last few decades many attempts have been made to use biocatalysts for the biotransformation of emerging contaminants in environmental matrices. Laccase, a multicopper oxidoreductase enzyme, has shown great potential in oxidizing a large number of phenolic and non-phenolic emerging contaminants. However, laccases and more broadly enzymes in their free form are biocatalysts whose applications in solution have many drawbacks rendering them currently unsuitable for large scale use. To circumvent these limitations, the enzyme can be immobilized onto carriers or entrapped within capsules; these two immobilization techniques have the disadvantage of generating a large mass of non-catalytic product. Insolubilization of the free enzymes as cross-linked enzymes (CLEAs) is found to yield a greater volume ratio of biocatalyst while improving the characteristics of the biocatalyst. Ultimately, novel techniques of enzymes insolubilization and stabilization are feasible with the combination of cross-linked enzyme aggregates (combi-CLEAs) and enzyme polymer engineered structures (EPESs) for the elimination of emerging micropollutants in wastewater. In this review, fundamental features of laccases are provided in order to elucidate their catalytic mechanism, followed by different chemical aspects of the immobilization and insolubilization techniques applicable to laccases. Finally, kinetic and reactor design effects for enzymes in relation with the potential applications of laccases as combi-CLEAs and EPESs for the biotransformation of micropollutants in wastewater treatment are discussed.

Elimination of bisphenol a and triclosan using the enzymatic system of autochthonous colombian forest fungi.

Bisphenol A (BPA) and triclosan (TCS) are known or suspected potential endocrine disrupting chemicals (EDCs) which may pose a risk to human health and have an environmental impact. Enzyme preparations containing mainly laccases, obtained from Ganoderma stipitatum and Lentinus swartzii, two autochthonous Colombian forest white rot fungi (WRF), previously identified as high enzyme producers, were used to remove BPA and TCS from aqueous solutions. A Box-Behnken factorial design showed that pH, temperature, and duration of treatment were significant model terms for the elimination of BPA and TCS. Our results demonstrated that these EDCs were extensively removed from 5 mg L(-1) solutions after a contact time of 6 hours. Ninety-four percent of TCS and 97.8% of BPA were removed with the enzyme solution from G. stipitatum; 83.2% of TCS and 88.2% of BPA were removed with the L. swartzii enzyme solution. After a 6-hour treatment with enzymes from G. stipitatum and L. swartzii, up to 90% of the estrogenic activity of BPA was lost, as shown by the yeast estrogen screen assay. 2,2-Azino-bis-(3-ethylthiazoline-6-sulfonate) (ABTS) was used as a mediator (laccase/mediator system) and significantly improved the laccase catalyzed elimination of BPA and TCS. The elimination of BPA in the absence of a mediator resulted in production of oligomers of molecular weights of 454, 680, and 906 amu as determined by mass spectra analysis. The elimination of TCS in the same conditions produced dimers, trimers, and tetramers of molecular weights of 574, 859, and 1146 amu. Ecotoxicological studies using Daphnia pulex to determine lethal concentration (LC50) showed an important reduction of the toxicity of BPA and TCS solutions after enzymatic treatments. Use of laccases emerges thus as a key alternative in the development of innovative wastewater treatment technologies. Moreover, the exploitation of local biodiversity appears as a potentially promising approach for identifying new efficient strains for biotechnological applications.

Laccase-Based CLEAs: Chitosan as a Novel Cross-Linking Agent.

Laccase from Coriolopsis Polyzona was insolubilized as cross-linked enzyme aggregates (CLEAs) for the first time with chitosan as the cross-linking agent. Concentrations between 0.01 and 1.867 g/L of chitosan were used and between 0.05 and 600 mM of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride. The laccase was precipitated using ammonium sulphate and cross-linked simultaneously. Specific activity and thermal stability of these biocatalysts were measured. Activities of up to 737 U/g were obtained when 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) was used as a substrate. Moreover, the stability of these biocatalysts was improved with regards to thermal degradation compared to free laccase when exposed to denaturing conditions of high temperature and low pH. The CLEAs stability against chemical denaturants was also tested but no significant improvement was detected. The total amount of ABTS to be oxidized during thermal degradation by CLEAs and free laccase was calculated and the insolubilized enzymes were reported to oxidize more substrate than free laccase. The formation conditions were analyzed by response surface methodology in order to determine an optimal environment for the production of efficient laccase-based CLEAs using chitosan as the cross-linking agent. After 24 hours of formation at pH 3 and at 4°C without agitation, the CLEAs exhibit the best specific activity.

Analysis and comparison of biotreatment of air polluted with ethanol using biofiltration and biotrickling filtration.

This study analyses the performance of ethanol biofiltration with percolation (biotrickling filtration, BTF) comparing to a conventional biofilter (biofiltration, BF). Two biofilters packed with clay balls were operated in a range of inlet concentrations of ethanol in the air varying from 0.47 to 2.36 g m(-3). For both the BF and BTF, the specific growth rate (mu) and the elimination capacity (EC) decreased with the ethanol inlet concentration, presenting a kinetic of substrate inhibition. A Haldane-type model was adjusted for both biofilters in order to model both EC and mu as a function of the ethanol inlet concentration in the gas. The maximum EC was similar for both biofilters, at around 46 g m(-3) h(-1), whereas the maximum mu was 0.0057 h(-1) for the BF and 0.0103 h(-1) for the BTF. The maximum of ethanol removed, occurred at the lowest inlet concentration of (0.47 gm(-3)), and reached 86% for the BF and 74% for the BTF.

Immobilization of laccase from the white rot fungus Coriolopsis polyzona and use of the immobilized biocatalyst for the continuous elimination of endocrine disrupting chemicals.

Laccase from the white rot fungus strain Coriolopsis polyzona was immobilized covalently on the diatomaceous earth support Celite R-633 using different strategies. A first methodology involved the sequential activation of the support surface with gamma-aminopropyltriethoxysilane followed by the reaction of the functionalized surface with glutaraldehyde (GLU) or glyoxal (GLY) and the immobilization of laccase on the activated surface. Another strategy tested the simultaneous internal cross-linking of the protein with GLU or GLY and the immobilization of the laccase on the silanized surface. Finally, these two strategies were modified to test the impact of the concomitant addition of bovine serum albumin (BSA) as a stabilizing agent during the immobilization steps. The highest laccase activity and the greatest degree of activity recovery (tested using 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) as the substrate) were achieved by the sequential immobilization procedure using GLU as the cross-linking agent. The solid catalysts featuring internal cross-linking of the protein showed significantly higher stability against several denaturants. The Michaelis-Menten kinetic parameters with respect to ABTS revealed a higher affinity for this substrate in the case of the sequential procedure compared to the simultaneous approach. The biocatalyst formed using GLU in the sequential procedure was applied in a packed bed reactor for the continuous treatment of 5 mg l(-1) solutions of the endocrine disrupting chemicals (EDCs) nonylphenol (NP), bisphenol A (BPA) and triclosan (TCS) through repeated batch treatments. All of these EDCs could be eliminated at a contact time of less than 200 min by using, respectively, 3.75 units (U) of laccase activity for BPA and TCS and 1.88 U for NP. These performances of elimination were maintained over five consecutive treatment cycles using the same biocatalyst. This system could also remove these EDCs from 100 mg l(-1) solutions. The Michaelis-Menten kinetic parameters with respect to these chemicals showed a decreasing affinity of the solid biocatalyst for NP, TCS and BPA in that order.

Utilization of cross-linked laccase aggregates in a perfusion basket reactor for the continuous elimination of endocrine-disrupting chemicals.

A perfusion basket reactor (BR) was developed for the continuous utilization of insolubilized laccase as cross-linked enzyme aggregates (CLEAs). The BR consisted of an unbaffled basket made of a metallic filtration module filled with CLEAs and continuously agitated by a 3-blade marine propeller. The agitation conditions influenced both the apparent laccase activity in the reactor and the stability of the biocatalyst. Optimal laccase activity was obtained at a rotational speed of 12.5 rps and the highest stability was reached at speeds of 1.7 rps or lower. The activity and stability of the biocatalyst were affected drastically upon the appearance of vortices in the reaction medium. This reactor was used for the continuous elimination of the endocrine disrupting chemicals (EDCs) nonylphenol (NP), bisphenol A (BPA), and triclosan (TCS). Optimization of EDC elimination by laccase CLEAs as a function of temperature and pH was achieved by response surface methodology using a central composite factorial design. The optimal conditions of pH and temperature were, respectively, 4.8 and 40.3 degrees C for the elimination of p353NP (a branched isomer of NP), 4.7 and 48.0 degrees C for BPA, and 4.9 and 41.2 degrees C for TCS. Finally, the BR was used for the continuous elimination of these EDCs from a 5 mg L(-1) aqueous solution using 1 mg of CLEAs at pH 5 and room temperature. Our results showed that at least 85% of these EDCs could be eliminated with a hydraulic retention time of 325 min. The performances of the BR were quite stable over a 7-day period of continuous treatment. Furthermore, this system could eliminate the same EDCs from a 100 mg L(-1) solution. Finally, a mathematical model combining the Michaelis-Menten kinetics of the laccase CLEAs and the continuous stirred tank reactor behavior of the BR was developed to predict the elimination of these xenobiotics.

Kinetics of microbial growth and biodegradation of methanol and toluene in biofilters and an analysis of the energetic indicators.

The kinetics of microbial growth and the biodegradation of methanol and toluene in (a) biofilters (BFs), and (b) biotrickling filters (BTFs), packed with inert materials, has been studied and analyzed. The specific growth rate, mu, for the treatment of methanol was 0.037h(-1) for a wide range of operating conditions. In the BF, mu was found to be a function of the methanol and toluene concentrations in the biofilm. In the BF used for treating methanol, mu was found to be affected by (1) the nitrogen concentration present in the nutrient solution, and (2) the kind of packing material employed. The kinetics of the methanol and toluene biodegradations were also analyzed using "mixed order" models. A Michaelis-Menten model type provided a good fit for the elimination capacity (EC) of the BTF treating methanol, while a Haldane model type provided a good fit to the EC of the BF treating methanol and toluene. The carbon dioxide production rate was related to the packed bed temperature and the content of the volatile solids within the biofilm. For the BF, the ratio of temperature/carbon dioxide production rate (PCO(2)) was 0.024 degrees C per unit of PCO(2), and for the BTF it was 0.15 degrees C per unit of PCO(2).

Preparation and characterization of cross-linked laccase aggregates and their application to the elimination of endocrine disrupting chemicals.

Laccase from the white rot fungus Coriolopsis polyzona was immobilized for the first time through the formation of cross-linked enzyme aggregates (CLEAs). Laccase CLEAs were produced by using 1000g of polyethylene glycol per liter of enzyme solution as precipitant and 200muM of glutaraldehyde as a cross-linking agent. These CLEAs had a laccase activity of 148Ug(-1) and an activity recovery of 60.2% when using 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) as substrate. CLEAs formed by co-aggregation with bovine serum albumin (BSA) as a stabilizer showed lower laccase activity and affinity for ABTS than those without BSA. The CLEAs co-aggregated with BSA showed higher residual activity against a protease, NaN(3), EDTA, methanol and acetone. The thermoresistance was higher for CLEAs than for free laccase and also higher for CLEAs co-aggregated with BSA than for simple CLEAs when tested at a pH of 3 and a temperature of 40 degrees C. Finally, laccase CLEAs were tested for their capacity to eliminate the known or suspected endocrine disrupting chemicals (EDCs) nonylphenol, bisphenol A and triclosan in a fluidized bed reactor. A 100-ml reactor with 0.5mg of laccase CLEAs operated continuously at a hydraulic retention time of 150min at room temperature and pH 5 could remove all three EDCs from a 5mgl(-1) solution.

Elimination of endocrine disrupting chemicals nonylphenol and bisphenol A and personal care product ingredient triclosan using enzyme preparation from the white rot fungus Coriolopsis polyzona.

The biocatalytic elimination of the endocrine disrupting chemicals (EDC) nonylphenol (NP) and bisphenol A (BPA) and the personal care product ingredient triclosan (TCS) by the enzyme preparation from the white rot fungus Coriolopsis polyzona was investigated. Analysis of variance methodology showed that the pH and the temperature are statistically significant factors in the removal of NP, BPA and TCS. The elimination of NP and TCS was best at a temperature of 50 degrees C and the disappearance of BPA at 40 degrees C, whereas the most suitable pH for all three micropollutants was 5. After a 4-h treatment of the three target compounds at concentrations of 5 mg l(-1) all of the NP and BPA were eliminated. In the case of TCS, 65% was removed after either a 4 or an 8-h treatment. The utilisation of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) in the laccase/mediator system significantly increased the efficiency of the enzymatic treatment. The elimination of NP and BPA was directly associated with the disappearance of the estrogenic activity. Mass spectrometry analysis showed that the enzymatic treatment produced high molecular weight metabolites through a radical polymerization mechanism of NP, BPA and TCS. These oligomers were produced through the formation of C-C or C-O bonds. The polymerization of NP produced dimers, trimers, tetramers and pentamers which had molecular weights of 438, 656, 874 and 1092 amu respectively. The polymerization of BPA produced dimers, trimers and tetramers which had molecular weights of 454, 680 and 906 amu. Finally, the polymerization of TCS produced dimers, trimers and tetramers which had molecular weights of 574, 859 and 1146 amu.