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The enzyme carbonic anhydrase (CA) has gainned considerable attention from the literature and the industry in the context of CO2 capture. CA immobilization in gas-liquid membrane contactors, and more specifically, on poly(ionic liquid) (PIL) composite membranes has been demonstrated to be a potential strategy to facilitate its industrial implementation. These membranes were comprised of a PIL layer coating on a porous hydrophobic polymeric support. In this work, the composition of the PIL layer was tuned by anion exchange to yield a variety of enzyme carriers. The following anions were compared: bromide [Br], acetate [Ac], tetrafluoroborate [BF4], and bis(trifluoromethylsulfonyl)imide [NTf2]. The surface morphology, chemistry, and properties of these composite membranes were characterized by SEM, EDX, ATR-FTIR, and water contact angle. The activity of the different biocatalytic composite membranes was determined by the p-nitrophenyl acetate hydrolysis model reaction. It was found that the anion exchange salts had a detrimental effect on the immobilized enzyme activity. In light of these results, the enzyme immobilization step was conducted after anion exchange. The resulting biocatalytic membranes displayed slight differences in immobilized enzyme activities and thermal stabilities following the order [Br]>[BF4]>[Ac]>[NTf2] and [BF4]>[Br]≈[Ac]>[NTf2], respectively. The differences were more pronounced and detrimental for the most hydrophobic anion, [NTf2]. Parallel trends were noted when the membranes were tested for CO2 absorption in a gas-liquid membrane contactor set-up suggesting that the CO2 mass transfer is strongly influenced by the activity of the immobilized enzymes. In addition, the effect of the absorption conditions, i.e., solvent flow rate, solvent saturation, and solvent concentration were evaluated. Under the best conditions, the novel biocatalytic membranes outperformed the commercial PVDF support by about a factor of 4 in terms of overall mass transfer coefficient. Such improvement would result in significant reductions in the required membrane area to capture CO2 by a gas-liquid membrane contactor.

This study presents an overview of and innovations in reverse osmosis (RO) membrane processes for rejecting charged metal ions in wastewater in relation to the main problems associated with purification methods. It also explains the emergence of nanomaterials and the different methods applied for RO membrane modification to improve performance. Membrane regeneration and retentate management are also considered. The study concludes with an economic feasibility study for the industrial scale-up of the methodology.

Fluoride is a concern for human health at high concentrations, but it is also a valuable compound with multiple applications. Thus, having a system that gives the opportunity to remove and recover this valuable element from water is highly interesting. Reverse osmosis (RO) is a promising technology in the removal of fluoride from water. Nevertheless, the residual retentate highly concentrated in fluoride is still a concern. The aim of this study was to evaluate the performance of an integrated process consisting of RO and membrane crystallization to remove fluoride from water and to recover it as a pure fluoride salt. Pure water permeability and fluoride rejection of a commercial RO membrane was tested under different conditions. In addition, the performance of an osmotic membrane crystallization setup was evaluated, considering the effect caused by the flow rates and the concentration of both the feed and the osmotic solution on the mass transfer coefficient. The crystallization process allowed the production of pure NaF crystals with octahedral morphology with a face-centered cubic crystal system.

This study explores the co-crystallization between the drug praziquantel (PZQ) and the nutraceutical curcumin (CU). The investigation revealed two novel solid forms: a cocrystal solvate with ethyl acetate and a non-solvated cocrystal. This novel drug–nutraceutical cocrystal is a praziquantel–curcumin (2:1) cocrystal. The cocrystal solvate has ethyl acetate molecules occupying the voids with minimal interactions within the crystal lattice. The application of heat treatment induces solvent removal and prompts the transition to the non-solvated cocrystal, as highlighted by variable-temperature X-ray powder diffraction (VT-XRPD). Thermal analyses demonstrate the stability of the cocrystal solvate up to approximately 100 °C, beyond which it transforms into the non-solvated phase, which eventually melts at 130 °C.

This study evaluates the performance of commercial reverse osmosis (RO) and nanofiltration (NF) membranes for the removal of metal ions from synthetic water and surface water carried from the north-west of Lake Tanganyika in the city of Uvira, in the east of the Democratic Republic of Congo. Metal ion analyses were performed by the standardized ICP-MS and ICP-OES methods. The RO membrane showed higher metal ion rejection in high-concentration solutions (synthetic samples) prepared in the laboratory as well as in low-concentration samples from real raw water collected near Lake Tanganyika. Rejection levels were higher than 98% for Cr3+, Pb2+, Cd2+, As3+, Ni2+, and Sb+3 ions in the synthetic solutions, and 99.2, 98.8, 98.6, 99.2, 98.4, and 98.8%, respectively, in the real samples. The concentrations of metals in the permeate varied depending on the feed concentration and were 0.15 to 1.02 mg/L, 0.33 to 22 mg/L, and 0.11 to 22 mg/L in RO, NF90, and NF270 membranes, respectively. Regarding the NF membranes, the rejection of Cr, Ni, and Cd ions was interesting: 98.2, 97.8, and 92.3%, respectively. However, it was lower for Pb, As, and Sb ions: 76.9, 52.5 and 64.1%, respectively. The flux of NF was 329 to 375 L/m2.h, much higher than for RO membranes, which had a flux of 98 to 132 L/m2.h. The studied membranes are thus a feasible solution to remove the studied metals from real water sources at low concentrations since they meet the standards of the World Health Organization on specific values assigned to chemicals from industrial sources and human habitation areas where these ions are present in drinking water.

Pyruvic acid is an organic compound used in various fields (e.g., the pharmaceutical, cosmetic, food, and chemical industries) and subject to constantly growing demand. Pyruvic acid is liquid at room temperature, rendering manipulation less straightforward. Furthermore, in the liquid phase, pyruvic acid is air-sensitive. We here present a multi-component crystal engineering strategy to render pyruvic acid solid under ambient conditions, focusing on cocrystallization and salt formation. Out of 73 screened cocrystal and salt formers, eight were found to form novel crystalline forms with pyruvic acid. Four of these were studied in detail, with pyruvic acid stable in a solid phase at temperatures up to 120 °C. These results illustrate the effectiveness of cocrystallization as a tool to convert unstable liquid compounds into stable crystalline solid forms.

Antisolvent crystallization is commonly used in the formation of heat-sensitive compounds as it is the case for most active pharmaceutical ingredients. Membranes have the ability to control the antisolvent mass transfer to the reaction medium, providing excellent mixing that inhibits the formation of local supersaturations responsible for the undesired properties of the resulting crystals. Still, optimization of the operating conditions is required. This work investigates the impact of solution velocity, the effect of antisolvent composition, the temperature and gravity, using glycine-water-ethanol as a model crystallization system, and polypropylene flat sheet membranes. Results proved that in any condition, membranes were consistent in providing a narrow crystal size distribution (CSD) with coefficient of variation (CV) in the range of 0.5–0.6 as opposed to 0.7 obtained by batch and drop-by-drop crystallization. The prism-like shape of glycine crystals was maintained as well, but slightly altered when operating at a temperature of 35 °C with the appearance of smoother crystal edges. Finally, the mean crystal size was within 23 to 40 µm and did not necessarily follow a clear correlation with the solution velocities or antisolvent composition, but increased with the application of higher temperature or gravity resistance. Besides, the monoclinic form of α-glycine was perfectly maintained in all conditions. The results at each condition correlated directly with the antisolvent transmembrane flux that ranged between 0.0002 and 0.001 kg/m2. s. In conclusion, membrane antisolvent crystallization is a robust solution offering consistent crystal properties under optimal operating conditions.

Metal–organic frameworks (MOFs), a sub-group of porous crystalline materials, have been receiving increasing attention for gas separation and pervaporation because of their high thermal and chemical stability, narrow window sizes, as well as tuneable structural, physical, and chemical properties. In this review, we comprehensively discuss developments in the formation of continuous MOF membranes for gas separation and pervaporation. Additionally, the application performance of continuous MOF membranes in gas separation and pervaporation are analysed. Lastly, some perspectives for the future application of continuous MOF membranes for gas separation and pervaporation are given.

A new crystallization process for sodium bicarbonate (NaHCO3) was studied, proposing the use of osmotic membrane distillation crystallization. Crystallization takes place due to the saturation of the feed solution after water evaporation on the feed side, permeating through the membrane pores to the osmotic side. The process operational parameters, i.e., feed and osmotic velocities, feed concentration, and temperature were studied to determine the optimal operating conditions. Regarding the feed and osmotic velocities, values of 0.038 and 0.0101 m/s, respectively, showed the highest transmembrane flux, i.e., 4.4 × 10−8 m3/m2·s. Moreover, study of the temperature variation illustrated that higher temperatures have a positive effect on the size and purity of the obtained crystals. The purity of the crystals obtained varied from 96.4 to 100% In addition, the flux changed from 2 × 10−8 to 7 × 10−8 m3/m2·s with an increase in temperature from 15 to 40 °C. However, due to heat exchange between the feed and the osmotic solutions, the energy loss in osmotic membrane distillation crystallization is higher at higher temperatures.

Mangrove sediments from New Caledonia were screened for xylanase sequences. One enzyme was selected and characterized both biochemically and for its industrial potential. Using a specific cDNA amplification method coupled with a MiSeq sequencing approach, the diversity of expressed genes encoding GH11 xylanases was investigated beneath Avicenia marina and Rhizophora stylosa trees during the wet and dry seasons and at two different sediment depths. GH11 xylanase diversity varied more according to tree species and season, than with respect to depth. One complete cDNA was selected (OFU29) and expressed in Pichia pastoris. The corresponding enzyme (called Xyn11-29) was biochemically characterized, revealing an optimal activity at 40–50 °C and at a pH of 5.5. Xyn11-29 was stable for 48 h at 35 °C, with a half-life of 1 h at 40 °C and in the pH range of 5.5–6. Xyn11-29 exhibited a high hydrolysis capacity on destarched wheat bran, with 40% and 16% of xylose and arabinose released after 24 h hydrolysis. Its activity on wheat straw was lower, with a release of 2.8% and 6.9% of xylose and arabinose, respectively. As the protein was isolated from mangrove sediments, the effect of sea salt on its activity was studied and discussed.

Following the concept of the holobiont, insect-microbiota interactions play an important role in insect biology. Many examples of host-associated microorganisms have been reported to drastically influence insect biological processes such as development, physiology, nutrition, survival, immunity, or even vector competence. While a huge number of studies on insect-associated microbiota have focused on bacteria, other microbial partners including fungi have been comparatively neglected. Yeasts, which establish mostly commensal or symbiotic relationships with their host, can dominate the mycobiota of certain insects. This review presents key advances and progress in the research field highlighting the diversity of yeast communities associated with insects, as well as their impact on insect life-history traits, immunity, and behavior.

Mosquitoes are considered one of the most important threats worldwide due to their ability to vector pathogens. They are responsible for the transmission of major pathogens such as malaria, dengue, zika, or chikungunya. Due to the lack of treatments or prophylaxis against many of the transmitted pathogens and an increasing prevalence of mosquito resistance to insecticides and drugs available, alternative strategies are now being explored. Some of these involve the use of microorganisms as promising agent to limit the fitness of mosquitoes, attract or repel them, and decrease the replication and transmission of pathogenic agents. In recent years, the importance of microorganisms colonizing the habitat of mosquitoes has particularly been investigated since they appeared to play major roles in their development and diseases transmission. In this issue, we will synthesize researches investigating how microorganisms present within water habitats may influence breeding site selection and oviposition strategies of gravid mosquito females. We will also highlight the impact of such microbes on the fate of females’ progeny during their immature stages with a specific focus on egg hatching, development rate, and larvae or pupae survival.

The functional diversity of the New Caledonian mangrove sediments was examined, observing the distribution of fungal dye-decolorizing peroxidases (DyPs), together with the complete biochemical characterization of the main DyP. Using a functional metabarcoding approach, the diversity of expressed genes encoding fungal DyPs was investigated in surface and deeper sediments, collected beneath either Avicennia marina or Rhizophora stylosa trees, during either the wet or the dry seasons. The highest DyP diversity was observed in surface sediments beneath the R. stylosa area during the wet season, and one particular operational functional unit (OFU1) was detected as the most abundant DyP isoform. This OFU was found in all sediment samples, representing 51–100% of the total DyP-encoding sequences in 70% of the samples. The complete cDNA sequence corresponding to this abundant DyP (OFU 1) was retrieved by gene capture, cloned, and heterologously expressed in Pichia pastoris. The recombinant enzyme, called DyP1, was purified and characterized, leading to the description of its physical–chemical properties, its ability to oxidize diverse phenolic substrates, and its potential to decolorize textile dyes; DyP1 was more active at low pH, though moderately stable over a wide pH range. The enzyme was very stable at temperatures up to 50 °C, retaining 60% activity after 180 min incubation. Its ability to decolorize industrial dyes was also tested on Reactive Blue 19, Acid Black, Disperse Blue 79, and Reactive Black 5. The effect of hydrogen peroxide and sea salt on DyP1 activity was studied and compared to what is reported for previously characterized enzymes from terrestrial and marine-derived fungi.

Ionic liquids have attracted the attention of the industry and research community as versatile solvents with unique properties, such as ionic conductivity, low volatility, high solubility of gases and vapors, thermal stability, and the possibility to combine anions and cations to yield an almost endless list of different structures. These features open perspectives for numerous applications, such as the reaction medium for chemical synthesis, electrolytes for batteries, solvent for gas sorption processes, and also membranes for gas separation. In the search for better-performing membrane materials and membranes for gas and vapor separation, ionic liquids have been investigated extensively in the last decade and a half. This review gives a complete overview of the main developments in the field of ionic liquid membranes since their first introduction. It covers all different materials, membrane types, their preparation, pure and mixed gas transport properties, and examples of potential gas separation applications. Special systems will also be discussed, including facilitated transport membranes and mixed matrix membranes. The main strengths and weaknesses of the different membrane types will be discussed, subdividing them into supported ionic liquid membranes (SILMs), poly(ionic liquids) or polymerized ionic liquids (PILs), polymer/ionic liquid blends (physically or chemically cross-linked ‘ion-gels’), and PIL/IL blends. Since membrane processes are advancing as an energy-efficient alternative to traditional separation processes, having shown promising results for complex new separation challenges like carbon capture as well, they may be the key to developing a more sustainable future society. In this light, this review presents the state-of-the-art of ionic liquid membranes, to analyze their potential in the gas separation processes of the future.

The growing expansion of mosquito vectors leads to the emergence of vector-borne diseases in new geographic areas and causes major public health concerns. In the absence of effective preventive treatments against most pathogens transmitted, vector control remains one of the most suitable strategies to prevent mosquito-borne diseases. Insecticide overuse raises mosquito resistance and deleterious impacts on the environment and non-target species. Growing knowledge of mosquito biology has allowed the development of alternative control methods. Following the concept of holobiont, mosquito-microbiota interactions play an important role in mosquito biology. Associated microbiota is known to influence many aspects of mosquito biology such as development, survival, immunity or even vector competence. Mosquito-associated microbiota is composed of bacteria, fungi, protists, viruses and nematodes. While an increasing number of studies have focused on bacteria, other microbial partners like fungi have been largely neglected despite their huge diversity. A better knowledge of mosquito-mycobiota interactions offers new opportunities to develop innovative mosquito control strategies. Here, we review the recent advances concerning the impact of mosquito-associated fungi, and particularly nonpathogenic fungi, on life-history traits (development, survival, reproduction), vector competence and behavior of mosquitoes by focusing on Culex, Aedes and Anopheles species.

Polymer-based CO2 selective membranes offer an energy efficient method to separate CO2 from flue gas. ‘Top-down’ polyelectrolytes represent a particularly interesting class of polymer materials based on their vast synthetic flexibility, tuneable interaction with gas molecules, ease of processability into thin films, and commercial availability of precursors. Recent developments in their synthesis and processing are reviewed herein. The four main groups of post-synthetically modified polyelectrolytes discern ionised neutral polymers, cation and anion functionalised polymers, and methacrylate-derived polyelectrolytes. These polyelectrolytes differentiate according to the origin and chemical structure of the precursor polymer. Polyelectrolytes are mostly processed into thin-film composite (TFC) membranes using physical and chemical layer deposition techniques such as solvent-casting, Langmuir-Blodgett, Layer-by-Layer, and chemical grafting. While solvent-casting allows manufacturing commercially competitive TFC membranes, other methods should still mature to become cost-efficient for large-scale application. Many post-synthetically modified polyelectrolytes exhibit outstanding selectivity for CO2 and some overcome the Robeson plot for CO2/N2 separation. However, their CO2 permeance remain low with only grafted and solvent-casted films being able to approach the industrially relevant performance parameters. The development of polyelectrolyte-based membranes for CO2 separation should direct further efforts at promoting the CO2 transport rates while maintaining high selectivities with additional emphasis on environmentally sourced precursor polymers.

A comparison between a traditional packed column and a novel membrane contactor used for CO2 absorption with carbonate production is addressed in this paper. Membrane technology is generally characterized by a lower energy consumption, it offers an independent control of gas and liquid streams, a known interfacial area and avoids solvent dragging. Those advantages make it a potential substitute of conventional absorption towers. The effect of the concentration and the flow rates of both the flue gas (10–15% of CO2) and the alkaline sorbent (NaOH, NaOH/Na2CO3) on the variation of the species present in the system, the mass transfer coefficient, and the CO2 removal efficiency was evaluated. Under the studied operation conditions, the membrane contactor showed very competitive results with the conventional absorption column, even though the highest mass transfer coefficient was found in the latter technology. In addition, the membrane contactor offers an intensification factor higher than five due to its compactness and modular character.

Turning carbon dioxide (CO2) into fuels and chemicals using chemical, photochemical, electrochemical, and enzymatic methods could be used to recycle large quantities of carbon. The enzymatic method, which is inspired by cellular CO2 metabolism, has attracted considerable attention for efficient CO2 conversion due to improved selectivity and yields under mild reaction conditions. In this review, the research progress of green and potent enzymatic conversion of CO2 into useful fuels and chemicals was discussed. Furthermore, applications of the enzymatic conversion of CO2 to assist in CO2 capture and sequestration were highlighted. A summary including the industrial applications, barriers, and some perspectives on the research and development of the enzymatic approach to convert CO2 were introduced.

It has been proven that a versatile bio-glue, polydopamine, can firmly bind TiO2 (titanium dioxide) nanoparticles on thin film composite (TFC) membranes. In this work, the anti-fouling behaviour of this novel polydopamine-TiO2-modified membrane is evaluated, based on the static bovine serum albumin (BSA) surface adhesion of the membranes and the relative flux decline. The results show that the anti-fouling performance of this new membrane is significantly improved in dark conditions when compared with the neat TFC membrane and the membranes only modified by polydopamine or TiO2. When filtrating a 0.5 g·L−1 BSA solution in dark conditions, the flux of the polydopamine-TiO2-modified membrane remains constant, at 95% of its pure water flux after 30 min filtration for 8 h of the experiment. This indicates a significant increase in anti-fouling performance when compared to the 25% flux decline observed for the neat TFC membrane, and to the 15% flux decline of those only modified by polydopamine or TiO2. This remarkable anti-fouling behaviour is attributed to an improved and uniform hydrophilicity, due to the presence of TiO2 and to the regular nanosized papillae structure of the polydopamine-TiO2 coating. Furthermore, since dopamine-modified TiO2 has visible light-induced photocatalytic properties, the membrane’s photocatalytic performance was also tested in light conditions. However an increase of flux and decrease of retention were observed after 24 h of continuous illumination, indicating that light may also affect the top layer of the membrane.

Fungal secretory peroxidases mediate fundamental ecological functions in the conversion and degradation of plant biomass. Many of these enzymes have strong oxidizing activities towards aromatic compounds and are involved in the degradation of plant cell wall (lignin) and humus. They comprise three major groups: class II peroxidases (including lignin peroxidase, manganese peroxidase, versatile peroxidase and generic peroxidase), dye-decolorizing peroxidases, and heme-thiolate peroxidases (e.g. unspecific/aromatic peroxygenase, chloroperoxidase). Here, we have repeatedly observed a widespread expression of all major peroxidase groups in leaf and needle litter across a range of forest ecosystems (e.g. Fagus, Picea, Acer, Quercus, and Populus spp.), which are widespread in Europe and North America. Manganese peroxidases and unspecific peroxygenases were found expressed in all nine investigated forest sites, and dye-decolorizing peroxidases were observed in five of the nine sites, thereby indicating biological significance of these enzymes for fungal physiology and ecosystem processes. Transcripts of selected secretory peroxidase genes were also analyzed in pure cultures of several litter-decomposing species and other fungi. Using this information, we were able to match, in environmental litter samples, two manganese peroxidase sequences to Mycena galopus and Mycena epipterygia and one unspecific peroxygenase transcript to Mycena galopus, suggesting an important role of this litter- and coarse woody debris-dwelling genus in the disintegration and transformation of litter aromatics and organic matter formation.

Ultrafiltration (UF) systems have been used globally for treating water from resources including rivers, reservoirs, and lakes for the production of potable water in the past decade. UF membranes with a pore size of between 0.1 and 0.01 micrometres provide an effective barrier for bacteria, viruses, suspended particles, and colloids. The use of UF membrane technology in treating groundwater for the supply of potable water in the impoverished and rural village, Tshaanda (i.e., the study area) is demonstrated. The technical and administrative processes that are critical for the successful installation of the pilot plant were developed. Given the rural nature of Tshaanda, the cultural and traditional protocols were observed. Preliminary results of the water quality of untreated water and the permeate are presented. Escherichia coli in the untreated water during the dry season (i.e., June and July) was 2 cfu/100 ml and was 2419.2 cfu/100 ml) before UF. Following UF, it dramatically reduced to acceptable level (7 cfu/100 ml) which is within the WHO recommended level of

Plant biomass degradation in soil is one of the key steps of carbon cycling in terrestrial ecosystems. Fungal saprotrophic communities play an essential role in this process by producing hydrolytic enzymes active on the main components of plant organic matter. Open questions in this field regard the diversity of the species involved, the major biochemical pathways implicated and how these are affected by external factors such as litter quality or climate changes. This can be tackled by environmental genomic approaches involving the systematic sequencing of key enzyme-coding gene families using soil-extracted RNA as material. Such an approach necessitates the design and evaluation of gene family-specific PCR primers producing sequence fragments compatible with high-throughput sequencing approaches. In the present study, we developed and evaluated PCR primers for the specific amplification of fungal CAZy Glycoside Hydrolase gene families GH5 (subfamily 5) and GH11 encoding endo-β-1,4-glucanases and endo-β-1,4-xylanases respectively as well as Basidiomycota class II peroxidases, corresponding to the CAZy Auxiliary Activity family 2 (AA2), active on lignin. These primers were experimentally validated using DNA extracted from a wide range of Ascomycota and Basidiomycota species including 27 with sequenced genomes. Along with the published primers for Glycoside Hydrolase GH7 encoding enzymes active on cellulose, the newly design primers were shown to be compatible with the Illumina MiSeq sequencing technology. Sequences obtained from RNA extracted from beech or spruce forest soils showed a high diversity and were uniformly distributed in gene trees featuring the global diversity of these gene families. This high-throughput sequencing approach using several degenerate primers constitutes a robust method, which allows the simultaneous characterization of the diversity of different fungal transcripts involved in plant organic matter degradation and may lead to the discovery of complex patterns in gene expression of soil fungal communities.

Eukaryotic organisms play essential roles in the biology and fertility of soils. For example the micro and mesofauna contribute to the fragmentation and homogenization of plant organic matter, while its hydrolysis is primarily performed by the fungi. To get a global picture of the activities carried out by soil eukaryotes we sequenced 2×10,000 cDNAs synthesized from polyadenylated mRNA directly extracted from soils sampled in beech (Fagus sylvatica) and spruce (Picea abies) forests. Taxonomic affiliation of both cDNAs and 18S rRNA sequences showed a dominance of sequences from fungi (up to 60%) and metazoans while protists represented less than 12% of the 18S rRNA sequences. Sixty percent of cDNA sequences from beech forest soil and 52% from spruce forest soil had no homologs in the GenBank/EMBL/DDJB protein database. A Gene Ontology term was attributed to 39% and 31.5% of the spruce and beech soil sequences respectively. Altogether 2076 sequences were putative homologs to different enzyme classes participating to 129 KEGG pathways among which several were implicated in the utilisation of soil nutrients such as nitrogen (ammonium, amino acids, oligopeptides), sugars, phosphates and sulfate. Specific annotation of plant cell wall degrading enzymes identified enzymes active on major polymers (cellulose, hemicelluloses, pectin, lignin) and glycoside hydrolases represented 0.5% (beech soil)-0.8% (spruce soil) of the cDNAs. Other sequences coding enzymes active on organic matter (extracellular proteases, lipases, a phytase, P450 monooxygenases) were identified, thus underlining the biotechnological potential of eukaryotic metatranscriptomes. The phylogenetic affiliation of 12 full-length carbohydrate active enzymes showed that most of them were distantly related to sequences from known fungi. For example, a putative GH45 endocellulase was closely associated to molluscan sequences, while a GH7 cellobiohydrolase was closest to crustacean sequences, thus suggesting a potentially significant contribution of non-fungal eukaryotes in the actual hydrolysis of soil organic matter.