Surfactant-aided mycoremediation of soil contaminated with polycyclic aromatic hydrocarbons.

Remediation of persistent polycyclic aromatic hydrocarbons (PAHs) contaminated soil has become a major challenge in recent years. Further, conventional application of bioaugmentation strategies for PAHs remediation require continuous supply of microbial specific nutrients, which makes these processes less feasible. Hence, the present study focused on PAHs remediation using surfactants along with wood assisted fungal system in a microcosm set up. In this study, in absence of surfactants, a saturation in PAHs degradation was noted in bioaugmentation with wood assisted fungal system (BAW) with 61 ± 1.25% degradation, followed by bioaugmentation with free fungi system (BAF) (54 ± 0.46%). However, with addition of 1500 mg/L of surface-active compounds (SAC), a maximum PAHs degradation in BAW (100%) and BAF (86 ± 1.30%) strategies were noted on 21st day. Irrespective of the strategies, presence of SAC and rhamnolipids enhanced PAHs degradation by increasing the enzymes production in Trametes hirsuta when compared to Triton x-100 and sodium dodecyl sulphate (SDS). Among the detected PAHs, 100% degradation within 17 days was noted for naphthalene and acenaphthene in SAC-supplemented BAW system. Further, ecotoxicity analysis established showed the LC50 of sediment soil at 26.5 ± 0.24%, which was reduced by an average of 71% after soil remediation. Hence, the current microcosm system proved that the application of SAC with BAW enhanced the PAHs remediation rate, which supports its application in real time soil remediation.

Insolubilization of Tramates versicolor laccase as cross-linked enzyme aggregates for the remediation of trace organic contaminants from municipal wastewater.

The novelty of this study deals with the biocatalytic treatment of trace organic contaminants (TrOCs) from municipal wastewater by insolubilized laccase. Laccase from Trametes versicolor was aggregated by three-phase partitioning technique followed by cross-linking with glutaraldehyde to produce insolubilized laccase as cross-linked enzyme aggregates (CLEAs). The optimal conditions for CLEAs preparation include ammonium sulphate concentration of 83% (w/v), crude to t-butanol ratio of 1.00: 1.05 (v/v), pH 5.3, and glutaraldehyde concentration of 20 mM obtained via statistical design. The efficiency of insolubilization of the CLEAs laccase based on the kcat/km ratio was approximately 4.8-fold greater than that of free laccase. The developed CLEAs showed greater resistance to product inhibition mediated by ABTS than the free enzyme and exhibited excellent catalytic activity even after the tenth successive cycle. Further, free laccase and the synthesized CLEAs laccase were utilized to treat five analgesics, two NSAIDS, three antibiotics, two antilipemics, and three pesticides in the municipal wastewater. Under the batch process with operating conditions of pH 7.0 and 20 °C, 1000 U/L of CLEAs, laccase removed 11 TrOCs in the range of about 20-99%. However, the inactivated CLEAs only adsorbed 2-25% of TrOCs. It was observed that acetaminophen, mefenamic acid, trimethoprim, and metolachlor depicted almost complete removal with CLEAs laccase. The performance of CLEAs laccase in a perfusion basket reactor was tested for the removal of TrOCs from municipal wastewater.

Coimmobilized enzymes as versatile biocatalytic tools for biomass valorization and remediation of environmental contaminants - A review.

Due to stringent regulatory norms, waste processing faces confrontations and challenges in adapting technology for effective management through a convenient and economical system. At the global level, attempts are underway to achieve a green and sustainable treatment for the valorization of lignocellulosic biomass as well as organic contaminants in wastewater. Enzymatic treatment in the environmental aspect thrived on being the promising rapid strategy that appeased the aforementioned predicament. On that account, coimmobilization of various enzymes on single support enhances the catalytic activity ensuing operational stability with industrial applications. This review pivoted towards the coimmobilization of enzymes on diverse supports and their applications in biomass conversion to industrial value-added products and removal of contaminants in wastewater. The limelight of this study chronicles the unique breakthroughs in biotechnology for the production of reusable biocatalysts, which inculcating various enzymes towards the scope of environment application.

Transcriptomic analysis of polyaromatic hydrocarbon degradation by the halophilic fungus Aspergillus sydowii at hypersaline conditions.

Polycyclic aromatic hydrocarbons (PAHs) are among the most persistent xenobiotic compounds, with high toxicity effects. Mycoremediation with halophilic Aspergillus sydowii was used for their removal from a hypersaline medium (1 M NaCl). A. sydowii metabolized PAHs as sole carbon sources, resulting in the removal of up to 90% for both PAHs [benzo [a] pyrene (BaP) and phenanthrene (Phe)] after 10 days. Elimination of Phe and BaP was almost exclusively due to biotransformation and not adsorption by dead mycelium and did not correlate with the activity of lignin modifying enzymes (LME). Transcriptomes of A. sydowii grown on PAHs, or on glucose as control, both at hypersaline conditions, revealed 170 upregulated and 76 downregulated genes. Upregulated genes were related to starvation, cell wall remodelling, degradation and metabolism of xenobiotics, DNA/RNA metabolism, energy generation, signalling and general stress responses. Changes of LME expression levels were not detected, while the chloroperoxidase gene, possibly related to detoxification processes in fungi, was strongly upregulated. We propose that two parallel metabolic pathways (mitochondrial and cytosolic) are involved in degradation and detoxification of PAHs in A. sydowii resulting in intracellular oxidation of PAHs. To the best of our knowledge, this is the most comprehensive transcriptomic analysis on fungal degradation of PAHs.

Efficiencies of selected biotreatments for the remediation of PAH in diluted bitumen contaminated soil microcosms.

Unconventional oils such as diluted bitumen from oil sands differs from most of conventional oils in terms of physiochemical properties and PAHs composition. This raises concerns regarding the effectiveness of current remediation strategies and protocols originally developed for conventional oil. Here we evaluated the efficiency of different biotreatment approaches, such as fungi inoculation (bioaugmentation), sludge addition (bioaugmentation/biostimulation), perennial grasses plantation (phytoremediation) and their combinations as well as natural attenuation (as control condition), for the remediation of soil contaminated by synthetic crude oil (a product of diluted bitumen) in laboratory microcosms. We specifically monitored the PAHs loss percentage (alkylated PAHs and unsubstituted 16 EPA Priority PAHs), the residue of PAHs and evaluated the ecotoxicity of soil after treatment. All treatments were highly efficient with more than ~ 80% of ∑PAHs loss after 60 days. Distinctive loss efficiencies between light PAHs (≤ 3 rings, ~ 96% average loss) and heavy PAHs (4-6 rings, ~ 29% average loss) were observed. The lowest average PAHs residue (0.10 ± 0.02 mg·kg-1, for an initial concentration of 0.29 ± 0.12 mg·kg-1) was achieved with the "sludge-plants (grasses)" combination. Sludge addition was the only treatment that achieved significantly lower ecotoxicity (3% ± 4% of growth inhibition of L. sativa) than the control (natural attenuation, 13% ± 4% of inhibition). Sludge addition, grasses plantation and "sludge-fungi combination" treatments could result in lower PAH exposure (than other treatments) in post-treated soil when using the Canadian Soil Quality Guidelines for the protection of environmental and human health for potentially carcinogenic and other PAHs.

Utilization of biosolids for glucose oxidase production: A potential bio-fenton reagent for advanced oxidation process for removal of pharmaceutically active compounds.

The current work focuses on the production of glucose oxidase (GOD) in sterilized biosolid (BS) slurries containing BS and municipal wastewater effluent. Various parameters were optimized for maximizing the GOD production and the effects of biostimulation on GOD production was investigated by adding synthetic media components. The studies on inoculum characteristics at an inoculum age of 72 h and inoculum size of 20% (w/v) produced high GOD activities of around 6012 U/L in 25% (dw/v) BS media. Further, the effect of ultrasonication time was determined to release BS-bound GOD in order to maximize enzymes recovery. Using 1000 U/L of the BS-based GOD for 0.55 M glucose oxidation produced the maximum H2O2 concentration of 216 ppm. The produced H2O2 was utilized for bio-Fenton based advanced oxidation process for the partial removal of 15 pharmaceutically active compounds.

Evaluation of bio-fenton oxidation approach for the remediation of trichloroethylene from aqueous solutions.

In the present study, a Bio-Fenton oxidation approach for the removal of trichloroethylene (TCE) was developed through the optimization of enzyme-based hydrogen peroxide (H2O2) production from glucose. Glucose oxidase (GOD) was evaluated for the production of H2O2 and the optimized parameters were found to be the oxidation of 60 mM glucose by 1 mg mL-1 of GOD which yielded a conversion of 88.4% of glucose for subsequent utilization in the Bio-Fenton process. The biologically produced H2O2 was further used for the removal of TCE from groundwater samples by addition of ferrous ion to the reaction mixture. To determine the efficiency of hydroxyl radicals generated in Fenton reactions to remove TCE in a cost-efficient way, preliminary experiments such as classical Fenton and pseudo Bio-Fenton were carried out in aqueous solutions containing TCE in order to obtain the optimum conditions and then the efficiency of bio-Fenton was tested in groundwater. Based on these results, almost 97% of TCE was removed from aqueous solution within 30 min of classical Fenton under low pH condition (pH 3.0). While performing pseudo bio-Fenton experiments, the optimum conditions were determined to be pH 7.0 and 30 ppm H2O2 which yielded 61% removal of TCE in 30 min. Although a comparatively lower removal (30.2%) in groundwater containing TCE was achieved using Bio-Fenton process, it was at a neutral pH. The process was further enhanced by the addition of hydrogen peroxide after 3 h of reaction and a maximum of 48.4% removal of TCE was observed. Thus, this process will help to prevent the scavenging of the unstable H2O2 and can be employed in-situ.

Effect of soil organic matter (SOM) on the degradation of polycyclic aromatic hydrocarbons using Pleurotus dryinus IBB 903-A microcosm study.

Bioremediation of polycyclic aromatic hydrocarbons (PAHs) in soil matrix is often arbitrated by the presence of soil organic matter (SOM). The present study focuses on determining the effect of the soil organic content on the mycoremediation of two model PAHs, phenanthrene (Phe) and benzo(a) pyrene (BaP) through microcosm studies. On comparing various microcosm strategies, a maximum degradation of Phe (99%) and BaP (48.5%) in soil bioaugmented with Pleurotus dryinus IBB 903, followed by, biostimulation with the degradation of Phe (89.9%) and BaP (24.8%) were noted. On relating the degradation pattern with lignolytic enzyme cocktail production, a laccase activity of 108 U/Kg at day 21, aryl alcohol oxidase (411 U/Kg) and manganese peroxidase (52.2 U/Kg) at day 14 along with lignin peroxidase (481 U/Kg) at day 21 were noted in fungal augmented soils, which were comparatively higher than levels observed in the bio-stimulation. Investigating the impact of different concentration of SOM (3-12%), a maximum remediation of Phe by 100% at 9% SOM in days 28 and 58.19% for BaP at 12% SOM, respectively was exhibited. Further, the biosorption effect of PAHs in abiotic condition showed a positive correlation with the increase in SOM, with a maximum adsorption of 3.78% Phe, and 6.93% BaP. The results support that the nominal adsorption ability of SOM, and helps in enhancing the microbial growth, thereby improving their degradation potentials, when less than 6% of SOM was utilized. Overall, this work establishes the critical role of organic matter in the soil with reference by simultaneous stimulation and degradation capability in complete PAHs remediation.

Hollow silica microspheres as robust immobilization carriers.

Hollow silica microspheres provide an ideal solid support for enzyme immobilization. We tested one of the newest development, namely MATSPHERES®, a silica openwork material as a carrier for the covalent immobilization of enzymes used to synthesize bioactive compounds. Two model enzymes - ethylbenzene dehydrogenase and EL070 lipase - were considered. They belong to two different enzyme classes and catalyse reactions taking place in various environments (aqueous and non-aqueous, aerobic and anaerobic). The enzymes were immobilized by covalent bonds (via divinyl sulfone and glutaraldehyde) on new silica material. Effectiveness of immobilization processes on the spheres grafted with amine groups and on the analogues without functionalization was determined for both enzymes. Microspheres were characterized morphologically and also their mechanical stability was examined during exposure to varying physical conditions. It was shown that MATSPHERES® due to their openwork structure and relative stability under batch and flow conditions can be a competitive SBA support for enzyme immobilization and production of bioactive compounds.

Monitoring of prenatal exposure to organic and inorganic contaminants using meconium from an Eastern Canada cohort.

Evaluating in utero exposure to inorganic and multiclass organic contaminants is critical to better evaluate potential harmful effects on prenatal and postnatal development. The analysis of meconium, the first bowel discharge of the newborn, has been proposed as a non-invasive way to assess cumulative prenatal exposure. The aim of this study was to implement an analytical method for quantifying 72 targeted organic compounds, including pesticides, pharmaceutical compounds and daily life xenobiotics, in meconium in addition to selected elements (17 elements). We report initial monitoring results based on the analysis of 396 meconium samples from an Eastern Canada cohort (Quebec, Canada). Element contents in meconium were analysed by mass spectrometry after digestion in nitric acid and peroxide. Targeted organic compounds were extracted and purified from meconium samples by a solid-liquid extraction followed by a dispersive-SPE purification before tandem mass spectrometry analysis. Concentrations of targeted elements were within the range of concentration reported in European and US studies but were lower than concentrations found in a developing country cohort (i.e., Pb, Cd). Out of the 72 targeted organic compounds, 31 were detected at least once and 30 were quantified. Compounds with the highest frequency of detection were caffeine, detected in all samples (from 2.80 to 6186 ng g-1), followed by acetaminophen detected in 53% of the samples (up to ~402 µg g-1) and methyl paraben detected in 20% of the samples (up to ~10 µg g-1). Pesticides were detected in low frequencies (< 2%) and low concentration (< 35 ng g-1). Results show that meconium can be used to monitor prenatal exposure of foetus to a wide array of inorganic and organic contaminants.

Intracellular Enzymes Contribution to the Biocatalytic Removal of Pharmaceuticals by Trametes hirsuta.

The use of white rot fungi (WRF) for bioremediation of recalcitrant trace organic contaminants (TrOCs) is becoming greatly popular. Biosorption and lignin modifying enzymes (LMEs) are the most often reported mechanisms of action. Intracellular enzymes, such as cytochrome P450 (CYP450), have also been suggested to contribute. However, direct evidence of TrOCs uptake and intracellular transformation is lacking. The aim of this study was to evaluate the relative contribution of biosorption, extracellular LMEs activity, TrOCs uptake, and intracellular CYP450 on the removal of six nonsteroidal anti-inflammatories (NSAIs) by Trametes hirsuta. Results show that for most tested NSAIs, LMEs activity and biosorption failed to explain the observed removal. Most tested TrOCs are quickly taken up and intracellularly transformed. Fine characterization of intracellular transformation using ketoprofen showed that CYP450 is not the sole intracellular enzyme responsible for intracellular transformation. The contribution of CYP450 in further transformation of ketoprofen byproducts is also reported. These results illustrate that TrOCs transformation by WRF is a more complex process than previously reported. Rapid uptake of TrOCs and intracellular transformation through diverse enzymatic systems appears to be important components of WRF efficiency toward TrOCs.

Simple screening protocol for identification of potential mycoremediation tools for the elimination of polycyclic aromatic hydrocarbons and phenols from hyperalkalophile industrial effluents.

A number of fungal strains belonging to the ascomycota, basidiomycota and zygomycota genera were subjected to an in vitro screening regime to assess their ligninolytic activity potential, with a view to their potential use in mycoremediation-based strategies to remove phenolic compounds and polycyclic aromatic hydrocarbons (PAHs) from industrial wastewaters. All six basidiomycetes completely decolorized remazol brilliant blue R (RBBR), while also testing positive in both the guaiacol and gallic acid tests indicating good levels of lignolytic activity. All the fungi were capable of tolerating phenanthrene, benzo-α- pyrene, phenol and p-chlorophenol in agar medium at levels of 10 ppm. Six of the fungal strains, Pseudogymnoascus sp., Aspergillus caesiellus, Trametes hirsuta IBB 450, Phanerochate chrysosporium ATCC 787, Pleurotus ostreatus MTCC 1804 and Cadophora sp. produced both laccase and Mn peroxidase activity in the ranges of 200-560 U/L and 6-152 U/L, respectively, in liquid media under nitrogen limiting conditions. The levels of adsorption of the phenolic and PAHs were negligible with 99% biodegradation being observed in the case of benzo-α-pyrene, phenol and p-chlorophenol. The aforementioned six fungal strains were also found to be able to effectively treat highly alkaline industrial wastewater (pH 12.4). When this wastewater was supplemented with 0.1 mM glucose, all of the tested fungi, apart from A. caesiellus, displayed the capacity to remove both the phenolic and PAH compounds. Based on their biodegradative capacity we found T. hirsuta IBB 450 and Pseudogymnoascus sp., to have the greatest potential for further use in mycoremediation based strategies to treat wastestreams containing phenolics and PAHs.

Trace organic contaminants in lake waters: Occurrence and environmental risk assessment at the national scale in Canada.

Numerous contaminants are produced and used daily, a significant fraction ultimately finding their way into natural waters. However, data on their distribution in lakes is lacking. To address this gap, the presence of 54 trace organic contaminants (TrOCs), representative of various human activities, was investigated in the surface water of 290 lakes across Canada. These lakes ranged from remote to highly impacted by human activities. In 88% of the sampled lakes, contaminants were detected, with up to 28 detections in a single lake. The compounds most frequently encountered were atrazine, cotinine, and deethylatrazine, each of which was present in more than a third of the lakes. The range of detected concentrations was from 0.23 ng/L to about 2200 ng/L for individual compounds, while the maximum cumulative concentration exceeded 8100 ng/L in a single lake. A risk assessment based on effect concentrations for three aquatic species (Pimephales promelas, Daphnia magna, and Tetrahymena pyriformis) was conducted, revealing that 6% of lakes exhibited a high potential risk for at least one species. In 59% of lakes, some contaminants with potential sub-lethal effects were detected, with the detection of up to 17 TrOCs with potential impacts. The results of this work provide the first reference point for monitoring the evolution of contamination in Canadian lakes by TrOCs. They demonstrate that a high proportion of the sampled lakes bear an environmentally relevant anthropogenic chemical footprint.

Important Counteranion Effect on Adsorption Efficacity of Hydrogen Sulfide by Silver(I)-Dithione Coordination Polymers.

Four new coordination polymers (CPs) have been prepared and evaluated for their efficacy in adsorbing hydrogen sulfide. The reactions of the structurally flexible assembling dithione ligand, L, with different silver(I) salts lead to four new metal-organic coordination architectures (CPs I, III, V, and VIII) exhibiting either one- or two-dimensional networks. CP I, 2D-[(Ag2Cl2)L]n, exhibits a linear series of rhomboid (S)2Ag(µ2-Cl)2Ag(S)2 secondary building units (SBUs) where S is one of the thione functions of L, altogether forming a 2D-network. CP III, 2D-[(AgI)L]n, is built upon parallel staircase-shaped 1D-[Ag2(µ3-I)2]n SBUs bridged by S atoms of L that form a 2D-grid. CP V, 2D-[(AgL)(NO3)]n, presents parallel 1D-folded S-shaped [AgL]n+ chains linked by strong argentophilic Ag···Ag interactions, forming a 2D-scaffolding. CP VIII, 1D-[(Ag2L3)(Cr2O7)]n, shows 1D-zigzag [{Ag(η2-µ2,η-µ,µ-L)}2]n2n+ chains accompanied by Cr2O72- counteranions. The adsorption isotherms of H2S gas by these new CPs were examined and compared to those of related CPs [(Ag2Br2)L]n (II), [(AgCN)4L]n (IV), [(Ag2L)(CF3SO3)2]n (VI), and [(Ag2L)(NO3)(ClO4)]n (VII). Among the tested polymers, the 3D-CP IV featuring cyanide anions exhibits the highest adsorption capacity of the CPs studied in this work. In order to determine the reason for this marked difference, density functional theory (DFT) computations were used. All in all, it turns out that the electrostatic interactions (CN-···H-SH) are significantly stronger than the O-···H-SH ones. This investigation provides a valuable conceptual tool for other designs of CPs and MOFs having the purpose of capturing toxic gases.

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.

Characterization of laccases from Trametes hirsuta in the context of bioremediation of wastewater treatment plant effluent.

The bioremediation of pharmaceutical compounds contained in wastewater, in an ecological and sustainable way, is possible via the oxidative action of fungal laccases. The discovery of new fungal laccases with unique physico-chemical characteristics pushes researchers to identify suitable laccases for specific applications. The aim of this study is to purify and characterize laccase isoenzymes produced from the Trametes hirsuta IBB450 strain for the bioremediation of pharmaceutical compounds. Two main laccases mixtures were observed and purified in the extracts and were called Yn and Yg. Peptide fingerprinting analysis suggested that Yn was constituted mainly of laccase Q02497 and Yg of laccase A0A6M5CX58, respectively. Robustness tests, based on tolerance and stability, showed that both laccases were affected in a relatively similar way by salts (KCl, NaCl), organic solvents (ACN, MeOH), denaturing compounds (urea, trypsin, copper) and were virtually unaffected and stable in wastewater. Determination of kinetic constants (Michaelis (KM), catalytic constant (kcat) and kinetic efficiency (K=kcat/KM)) for the transformation of synthetic hormone 17α-ethynylestradiol and the anti-inflammatory agent diclofenac indicates a lower KM and kcat for laccase Yn but relative similar K constant compared to Yg. Synergistic effects were observed for the transformation of diclofenac, unlike 17α-ethynylestradiol. Transformation studies of 17α-ethynylestradiol at different temperatures (4 and 21 °C) indicate a transformation rate reduction of approximately 75-80% at 4 °C against 25% for diclofenac in less than an hour. Finally, the classification of laccases Yg and Yn into one of eight groups (group A-H) suggests that laccase Yg belongs to group A (constitutive laccase) and laccase Yn belongs to group B (inducible laccase).

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.

Genipin and glutaraldehyde based laccase two-layers immobilization with improved properties: New biocatalysts with high potential for enzymatic removal of trace organic contaminants.

This research proposes the preparation of a two-layer laccase biocatalyst using genipin or/and glutaraldehyde as cross-linking agents. The multilayer biocatalysts were prepared using different combinations of genipin and glutaraldehyde in the individual preparation of the first and second laccase layers. First, chitosan was treated with genipin or glutaraldehyde, followed by the immobilization of the first laccase layer to form a single-layer biocatalyst. Then, the immobilized laccases were coated once again with genipin or glutaraldehyde, and a new laccase layer was immobilized onto the system, resulting in the final two-layer biocatalyst. Compared to the single-layer biocatalysts, catalytic activity increased 1.7- and 3.4-fold when glutaraldehyde coating was used to prepare the second laccase layer. However, adding a second layer did not always produce more active biocatalysts, since the two-layer biocatalysts prepared with genipin (GenLacGenLac and GluLacGenLac) presented a decrease in activity of 65% and 28%, respectively. However, these two-layer biocatalysts prepared with genipin maintained 100% of their initial activity after 5 cycles of ABTS oxidation. Nevertheless, the two-layer, genipin-coated biocatalyst resulted in a higher removal of trace organic contaminants, since it removed 100% of mefenamic acid and 66% of acetaminophen, compared with the glutaraldehyde-coated biocatalyst, which removed 20% of mefenamic acid, and 18% of acetaminophen.

Cost-effective, scalable production of glucose oxidase using Casuarina equisetifolia biomass and its application in the bio-Fenton oxidation process for the removal of trace organic contaminants from wastewater.

This study focuses on using Casuarina equisetifolia biomass for pilot-scale glucose oxidase production from Aspergillus niger and its application in the removal of trace organic contaminants (TrOCs) from municipal wastewater through the bio-Fenton oxidation. The cost of glucose oxidase was 0.005 $/U, including the optimum production parameters, 10% biomass, 7% sucrose, 1% peptone, and 3% CaCO3 at 96 h with an enzyme activity of 670 U/mL. Optimized conditions for H2O2 were 1 M glucose, 100 U/mL glucose oxidase, and 120 mins of incubation, resulting in 544.3 mg/L H2O2. Thus, H2O2 produced under these conditions lead to bio-Fenton oxidation resulting in the removal of 36-92% of nine TrOCs in municipal wastewater at pH 7.0 in 360 mins. Therefore, this work establishes the cost-effective glucose oxidase-producing H2O2 as an attractive bioremediating agent to enhance the removal of TrOCs in wastewater at neutral pH.

Biological elimination of a high concentration of hydrogen sulfide from landfill biogas.

Hydrogen sulfide (H2S) is one of the main contaminants found in biogas, which is one of the end products of the anaerobic biodegradation of proteins and other sulfur-containing compounds in solid waste. The presence of H2S is one of the factors limiting the valorization of biogas. To valorize biogas, H2S must be removed. This study evaluated the performance of a pilot-scale biotrickling filter system on H2S removal from landfill biogas. The biotrickling filter system, which was packed with stainless-steel pall rings and inoculated with an H2S-oxidizing consortium, was designed to process 1 SCFM of biogas, which corresponds to an empty bed residence time (EBRT) of 3.9 min and was used to determine the removal efficiency of a high concentration of hydrogen sulfide from landfill biogas. The biofiltration system consisted of two biotrickling filters connected in series. Results indicate that the biofiltration system reduced H2S concentration by 94 to 98% without reducing the methane concentration in the outlet biogas. The inlet concentration of hydrogen sulfide, supplied to the two-phase bioreactor, was in the range of 900 to 1500 ppmv, and the air flow rate was 0.1 CFM. The EBRTs of the two biotrickling filters were 3.9 and 0.9 min, respectively. Approximately 50 ± 15.7 ppmv of H2S gas was detected in the outlet gas. The maximum elimination capacity of the biotrickling filter system was found to be 24 g H2S·m-3·h-1, and the removal efficiency was 94 ± 4.4%. During the biological process, the performance of the biotrickling filter was not affected when the pH of the recirculated liquid decreased to 2-3. The overall performance of the biotrickling filter system was described using a modified Michaelis-Menten equation, and the Ks and Vm values for the biosystem were 34.7 ppmv and 20 g H2S·m-3·h-1, respectively.