Noninvasive tool for optical online monitoring of individual biomass concentrations in a defined coculture.

Cocultures bear great potential in the conversion of complex substrates and process intensification, as well as, in the formation of unique components only available due to inter-species interactions. Dynamic data of coculture composition is necessary for understanding and optimizing coculture systems. However, most standard online determined parameters measure the sum of all species in the reactor system. The kinetic behavior of the individual species remains unknown. Up to now, different offline methods are available to determine the culture composition, as well as the online measurement of fluorescence of genetically modified organisms. To avoid any genetic modification, a noninvasive online monitoring tool based on the scattered light spectrum was developed for microtiter plate cultivations. To demonstrate the potential, a coculture consisting of the bacterium Lactococcus lactis and the yeast Kluyveromyces marxianus was cultivated. Via partial least squares regression of scattered light spectra, the online determination of the individual biomass concentrations without further sampling and analyses is possible. The results were successfully validated by a Coulter counter-analysis, taking advantage of the different cell sizes of both organisms. The findings prove the applicability of the new method to follow in detail the dynamics of a coculture.

Spathaspora passalidarum selected for resistance to AFEX hydrolysate shows decreased cell yield.

This study employed cell recycling, batch adaptation, cell mating and high-throughput screening to select adapted Spathaspora passalidarum strains with improved fermentative ability. The most promising candidate YK208-E11 (E11) showed a 3-fold increase in specific fermentation rate compared to the parental strain and an ethanol yield greater than 0.45 g/g substrate while co-utilizing cellobiose, glucose and xylose. Further characterization showed that strain E11 also makes 40% less biomass compared to the parental strain when cultivated in rich media under aerobic conditions. A tetrazolium agar overlay assay in the presence of respiration inhibitors, including rotenone, antimycin A, KCN and salicylhydroxamic acid elucidated the nature of the mutational events. Results indicated that E11 has a deficiency in its respiration system that could contribute to its low cell yield. Strain E11 was subjected to whole genome sequencing and an ∼11 kb deletion was identified; the open reading frames absent in strain E11 code for proteins with predicted functions in respiration, cell division and the actin cytoskeleton, and may contribute to the observed physiology of the adapted strain. Results of the tetrazolium overlay also suggest that cultivation on xylose affects the respiration capacity in the wild-type strain, which could account for its faster fermentation of xylose as compared to glucose. These results support our previous finding that S. passalidarum has highly unusual physiological responses to xylose under oxygen limitation.

Assessment of water samples with complex compositions using microalgal bioassay based on the community level physiological profiling (CLPP).

The ability to effectively characterize the response of microalgal communities to changes in water quality is limited. Earlier, a microalgal bioassay was developed based on community level physiological profiling (CLPP). The efficacy of this assay was evaluated using three wetland water samples, a surface water sample, and two wastewater samples (i.e. primary and secondary), all collected from southwestern Ontario, Canada. In addition, the assay was applied to untreated and activated carbon treated oil sand process water (OSPW). YT (Yeast Identification Test Panel) and Biolog plates were successfully utilized for defined microalgal community under both heterotrophic and mixotrophic growth conditions to characterize the changes in the defined microalgal community due to the changes in water type. It was found that, although the degrees of changes in the algal community varied, all tested water samples were distinguished under both growth regimes using principal component analysis (PCA). The variations in the algal community were caused by the differences of the water samples. The response of the assay due to changes in the algal community caused by different waters was found to be very sensitive and could be used to differentiate different water bodies. It further can be used to monitor temporal changes of water quality of the same water body.

Online measurement of viscosity for biological systems in stirred tank bioreactors.

One of the most critical parameters in chemical and biochemical processes is the viscosity of the medium. Its impact on mixing, as well as on mass and energy transfer is substantial. An increase of viscosity with reaction time can be caused by the formation of biopolymers like xanthan or by filamentous growth of microorganisms. In either case the properties of fermentation broth are changing and frequently non-Newtonian behavior are observed, resulting in major challenges for the measurement and control of mixing and mass transfer. This study demonstrates a method for the online determination of the viscosity inside a stirred tank reactor. The presented method is based on online measurement of heat transfer capacity from the bulk medium to the jacket of the reactor. To prove the feasibility of the method, fermentations with the xanthan producing bacterium Xanthomonas campestris pv. campestris B100 as model system were performed. Excellent correlation between offline measured apparent viscosity and online determined heat transfer capacity were found. The developed tool should be applicable to any other process with formation of biopolymers and filamentous growth. Biotechnol. Bioeng. 2017;114: 990-997. © 2016 Wiley Periodicals, Inc.

Consolidating biofuel platforms through the fermentative bioconversion of crude glycerol to butanol.

Economic realities for the rising industrial biofuel production have changed substantially during the low oil price period starting in the mid 2010's. Increased competition requires the sector to increase productivity through the reduction of low-value by-products and full utilization of all value and energy stored in their respective feedstock. Biodiesel is produced commercially from substrates such as animal fat and vegetable oil, generating approximately 10 wt% crude glycerol as its main, currently underutilized, by-product. This crude glycerol is contaminated with catalyst, soap, free fatty acids, glycerides and methyl esters; hence only a small fraction enters the existing glycerol markets, while the purification costs for the majority of crude glycerol are simply too high. However, this presents a unique opportunity to generate additional value. One technical possibility is to use crude glycerol as a carbon source for butanol production, a compound of higher value and energy, a potential additive for gasoline and diesel fuels and bulk chemical commodity. Conversion facilities could be co-located with biodiesel plants, utilizing established infrastructure and adding significant value and productivity to the existing biodiesel industry. This review focuses on the current activities geared towards the bioconversion of crude glycerol to butanol.

Geometric effects on non-DLVO forces: relevance for nanosystems.

In this paper, the surface element integration (SEI) method was used derive analytical force/potential versus distance profiles for two non-DLVO forces: Lewis acid-base and solvation forces. These forces are highly relevant in a variety of systems, from bacterial adhesion, nanoparticle suspension stability to atomic force microscopy (AFM) profiles. The SEI-derived expressions were compared with the more commonly utilized Derjaguin approximations in order to assess the effect of curvature on the resulting interaction for the test cases of sphere-flat plate and equally sized spheres. For acid-base interactions, the deviation was found to be significant for particles up to 40 nm in diameter for the conventionally used decay length (λ = 1 nm) for water. The resulting expressions show that accounting in curvature for acid-base interactions is important even for simple smooth geometric shapes, recovering the Derjaguin expression at smaller values of λ/R. These results allow for correction of the acid-base force/potential versus distance from the Derjaguin-derived expressions using simple functions of λ/R. Conversely, for the solvation force the deviation was far less significant due to the oscillatory nature of the potential damping out effects and the smaller order of magnitude range of the solvation decay length, indicating that for solvation forces the Derjaguin approximation is suitable for most conceivable cases.

Extrusion pretreatment of lignocellulosic biomass: a review.

Bioconversion of lignocellulosic biomass to bioethanol has shown environmental, economic and energetic advantages in comparison to bioethanol produced from sugar or starch. However, the pretreatment process for increasing the enzymatic accessibility and improving the digestibility of cellulose is hindered by many physical-chemical, structural and compositional factors, which make these materials difficult to be used as feedstocks for ethanol production. A wide range of pretreatment methods has been developed to alter or remove structural and compositional impediments to (enzymatic) hydrolysis over the last few decades; however, only a few of them can be used at commercial scale due to economic feasibility. This paper will give an overview of extrusion pretreatment for bioethanol production with a special focus on twin-screw extruders. An economic assessment of this pretreatment is also discussed to determine its feasibility for future industrial cellulosic ethanol plant designs.

Anaerobic digestibility of estrogens in wastewater sludge: effect of ultrasonic pretreatment.

BACKGROUND: Estrogenic compounds have been detected in the secondary effluents and in the biosolids from conventional wastewater treatment plants, which are not designed for their removal. Furthermore, existing limited studies on anaerobic digestibility of estrogens report conflicting results. The objective of the present work was to determine the fate and anaerobic digestibility of estrogenic compounds in various types of sludge including primary sludge (PS), waste activated sludge (WAS), and anaerobically digested sludge (seed). METHODS: Estrone (E1) and 17-ß estradiol (E2) were chosen as the model estrogenic compounds. Initially batch adsorption was conducted to determine the extent of adsorption and isotherm of E1 on various sludge. Thereafter, batch anaerobic digestion of E1 and E2 was conducted in various sludge using So/X ratio of 4 gCOD/gVSS in 250 ml bottles. The effect of earlier optimized ultrasonication dosage on the anaerobic digestion of E1 and E2 was also characterized. Estrogenicity of the digested samples was determined by the YES assay. RESULTS: Most of E1 and E2 was adsorbed on the biosolids and the Freundlich isotherm fitted the experimental data well. No anaerobic digestion of E1 and E2 was found in any of the sludge tested, and the estrogenicity of the sludge measured by YES assay increased during digestion due to the formation of E2 from E1 in a reduced environment. Ultrasonication decreased the initial mass of E1 and E2 by 20% in the sonicated digester as compared to control digester, however, there was no further decrease in E1 and E2 during digestion. CONCLUSIONS: Most of the estrogenic compounds partitioned onto the solids and remained there during digestion. Ultrasonication pretreatment reduced the estrogen burden for the digester due to advance oxidation, but no further removal of the estrogens occurred in the digester.

Environmental and Exergetic Analysis of Large-Scale Production of Citric Acid-Coated Magnetite Nanoparticles via Computer-Aided Process Engineering Tools.

Considering that functional magnetite (Fe3O4) nanoparticles with exceptional physicochemical properties can be highly applicable in different fields, scaling-up strategies are becoming important for their large-scale production. This study reports simulations of scaled-up production of citric acid-coated magnetite nanoparticles (Fe3O4-cit), aiming to evaluate the potential environmental impacts (PEIs) and the exergetic efficiency. The simulations were performed using the waste reduction algorithm and the Aspen Plus software. PEI and energy/exergy performance are calculated and quantified. The inlet and outlet streams are estimated by expanding the mass and energy flow, setting operating parameters of processing units, and defining a thermodynamic model for properties estimation. The high environmental performance of the production process is attributed to the low outlet rate of PEI compared to the inlet rate. The product streams generate low PEI contribution (-3.2 × 103 PEI/y) because of the generation of environmentally friendlier substances. The highest results in human toxicity potential (3.2 × 103 PEI/y), terrestrial toxicity potential (3.2 × 103 PEI/y), and photochemical oxidation potential (2.6 × 104 PEI/y) are attributed to the ethanol within the waste streams. The energy source contribution is considerably low with 27 PEI/y in the acidification potential ascribed to the elevated levels of hydrogen ions into the atmosphere. The global exergy of 1.38% is attributed to the high irreversibilities (1.7 × 105 MJ/h) in the separation stage, especially, to the centrifuge CF-2 (5.07%). The sensitivity analysis establishes that the global exergy efficiency increases when the performance of the centrifuge CF-2 is improved, suggesting to address enhancements toward low disposal of ethanol in the wastewater.

Feasibility of anaerobic digestion as a treatment for the aqueous pyrolysis condensate (APC) of birch bark.

Biooil produced via biomass pyrolysis includes an aqueous-acidic phase and a dense and rich organic phase. The aqueous phase has a low heating value and is considered a waste stream. In this study fractional condensation was employed to separate the liquid product of birch bark pyrolysis into an aqueous pyrolysis condensate (APC) and a dense biooil fraction. The APC contained high amounts (~100 g/kg) of acidic acid (AA) and was investigated for anaerobic digestion (AD). The AA in the APC could be converted to biogas, however, it contained elevated concentrations of microbial inhibitors (24 g/kg total phenolics). The inhibiting effect could be mitigated by acclimatization of the microbial population, which in turn converted some of the additional organics. The production of methane further improved with the addition of biochar to adsorb some of the inhibitors. The results imply that a waste product can be converted into a potential energy carrier.

Synthesis of FeO@SiO2-DNA core-shell engineered nanostructures for rapid adsorption of heavy metals in aqueous solutions.

Creating novel and innovative nanostructures is a challenge, aiming to discover nanomaterials with promising properties for environmental remediation. In this study, the physicochemical and adsorption properties of a heterogeneous nanostructure are evaluated for the rapid removal of heavy metal ions from aqueous solutions. Core-shell nanostructures are prepared using iron oxide cores and silica dioxide shells. The core is synthesized via the co-precipitation method and modified in situ with citric acid to grow a carboxyl layer. The shell was hydrolyzed/condensed and then functionalized with amine groups for ds-DNA condensation via electrostatic interaction. The characterization techniques revealed functional FeO@SiO2-DNA nanostructures with good crystallinity and superparamagnetic response (31.5 emu g-1). The predominant superparamagnetic nature is attributed to the citric acid coating. This improves the dispersion and stability of the magnetic cores through the reduction of the dipolar-dipolar interaction and the enhancement of the spin coordination. The rapid adsorption mechanism of FeO@SiO2-DNA was evaluated through the removal of Pb(ii), As(iii), and Hg(ii). A rapid adsorption rate is observed in the first 15 min, attributed to a heterogeneous chemisorption mechanism based on electrostatic interactions. FeO@SiO2-DNA shows higher adsorption efficiency of 69% for Pb(ii) removal compared to As(iii) (51%) and Hg(ii) (41%). The selectivity towards Pb(ii) is attributed to the similar acid nature to ds-DNA, where the ionic strength interaction provides good affinity and stability. The facile synthesis and rapid adsorption suggest a promising nanostructure for the remediation of water sources contaminated with heavy metal ions and can be extended to other complex molecules.

Comparison of Biobutanol Production Pathways via Acetone-Butanol-Ethanol Fermentation Using a Sustainability Exergy-Based Metric.

The incorporation of sustainability aspects into the design of chemical processes has been increasing since the last century. Hence, there are several proposed methodologies and indicators to assess chemical facilities through process analysis techniques. A comprehensive assessment involving economic, environmental, safety, and exergy parameters of two alternatives for butanol production from Manihot esculenta Crantz (cassava waste) is presented in this study. The modeling of process topologies involved using Aspen Plus software. Topology 1 generated a product flow rate of 316,477 t/y of butanol, while this value was 367,037 t/y for topology 2. Both processes used a feed flow of 3,131,439 t/y of biomass. This study used seven technical indicators to evaluate both alternatives, which include the return of investment, discounted payback period, global warming potential, renewability material index, inherent safety index, exergy efficiency, and exergy of waste ratio. Otherwise, this study implemented an aggregate index to assess overall sustainability performance. The results revealed that topology 2 presented higher economic normalized scores for evaluated indicators, but the most crucial difference between these designs came from the safety and exergetic indexes. Topology 1 and topology 2 obtained weighted scores equaling to 0.48 and 0.53; therefore, this study found that the second alternative gives a more sustainable design for butanol production under evaluated conditions.

Increased Butanol Yields through Cosubstrate Fermentation of Jerusalem Artichoke Tubers and Crude Glycerol by Clostridium pasteurianum DSM 525.

Clostridium pasteurianum DSM 525 can produce butanol, 1,3-propanediol, and ethanol from glycerol. The product distribution can be tilted toward butanol when adding butyric acid. The strain predominantly produces acetic and butyric acids when grown on saccharides. Hence, butyrate formed from saccharide conversion can be used to stimulate butanol production from glycerol under cosubstrate cultivation. The optimal cosubstrate ratio was determined, and under optimal conditions, a butanol yield and a productivity of 0.27 ± 0.01 gbutanol g-1 (glycerol + sugar) -1 and 0.74 ± 0.02 g L-1 h-1 were obtained. On the basis of these results, batch fermentation in a 5 L bioreactor was performed using Jerusalem artichoke hydrolysate (carbohydrate source) and crude glycerol (residue from biodiesel production) at the previously determined optimal condition. A butanol yield and a productivity of 0.28 ± 0.007 gbutanol g(glycerol+sugar) -1 and 0.55 ± 0.008 g L-1 h-1 were achieved after 27 h fermentation, indicating the suitability of those low-cost carbon sources as cosubstrates for butanol production via C. pasteurianum.

Clostridial conversion of corn syrup to Acetone-Butanol-Ethanol (ABE) via batch and fed-batch fermentation.

Corn syrup - a commercial product derived from saccharification of corn starch - was used to produce acetone-butanol-ethanol (ABE) by Clostridium spp. Screening of commercial Clostridium spp., substrate inhibition tests and fed-batch experiments were carried out to improve ABE production using corn syrup as only carbon source. The screening tests carried out in batch mode using a production media containing 50 g/L corn syrup revealed that C. saccharobutylicum was the best performer in terms of total solvent concentration (12.46 g/L), yield (0.30 g/g) and productivity (0.19 g/L/h) and it was selected for successive experiments. Concentration of corn syrup higher than 50 g/L resulted in no solvents production. Fed-batch fermentation improved ABE production with respect to batch fermentation: the butanol and solvent concentration increased up to 8.70 and 16.68 g/L, respectively. The study demonstrated the feasibility of producing solvents via ABE fermentation using corn syrup as a model substrate of concentrated sugar mixtures.

Fast media optimization for mixotrophic cultivation of Chlorella vulgaris.

Microalgae can accumulate large proportions of their dry cell weight as storage lipids when grown under appropriate nutrient limiting conditions. While a high ratio of carbon to nitrogen is often cited as the primary mode of triggering lipid accumulation in microalgae, fast optimization strategies to increase lipid production for mixotrophic cultivation have been difficult to developed due to the low cell densities of algal cultures, and consequently the limited amount of biomass available for compositional analysis. Response surface methodologies provide a power tool for assessing complex relationships such as the interaction between the carbon source and nitrogen source. A 15 run Box-Behnken design performed in shaker flasks was effective in studying the effect of carbon, nitrogen, and magnesium on the growth rate, maximum cell density, lipid accumulation rate, and glucose consumption rate. Using end-point dry cell weight and total lipid content as assessed by direct transesterification to FAME, numerical optimization resulted in a significant increase in lipid content from 18.5 ± 0.76% to 37.6 ± 0.12% and a cell density of 5.3 ± 0.1 g/L to 6.1 ± 0.1 g/L between the centre point of the design and the optimized culture conditions. The presented optimization process required less than 2 weeks to complete, was simple, and resulted in an overall lipid productivity of 383 mg/L·d.

Biodegradation of PCBs in two-phase partitioning bioreactors following solid extraction from soil.

This article demonstrates the feasibility of a novel process concept for the remediation of PCB contaminated soil. The proposed process consists of PCB extraction from soil using solid polymer beads, followed by biodegradation of the extracted PCBs in a solid-liquid two-phase partitioning bioreactor (TPPB), where PCBs are delivered from the polymer beads to the degrading organisms. The commercially available thermoplastic polymer Hytrel was used to extract Aroclor 1242 from contaminated artificial soil in bench scale experiments. Initial PCB contamination levels of 100 and 1,000 mg kg(-1) could be reduced to 32% +/- 1 to 41% +/- 7 of the initial value after 48 h mixing in the presence of a mobilizing agent at polymer-to-soil ratios of 1% (w/w) and 10% (w/w). The decrease of detectable PCBs in the soil was consistent with an increase of PCBs in the polymer beads. It was further shown that Aroclor 1242 could be delivered to the PCB degrading organism Burkholderia xenovorans LB400 in a solid-liquid TPPB via Hytrel beads. A total of 70 mg Aroclor 1242 could be degraded in a 1 L solid-liquid TPPB within 80 h of operation.

Enhancement of PCB degradation by Burkholderia xenovorans LB400 in biphasic systems by manipulating culture conditions.

Two-phase partitioning bioreactors (TPPBs) can be used to biodegrade environmental contaminants after their extraction from soil. TPPBs are typically stirred tank bioreactors containing an aqueous phase hosting the degrading microorganism and an immiscible, non-toxic and non-bioavailable organic phase functioning as a reservoir for hydrophobic compounds. Biodegradation of these compounds in the aqueous phase results in thermodynamic disequilibrium and partitioning of additional compounds from the organic phase into the aqueous phase. This self-regulated process can allow the delivery of large amounts of hydrophobic substances to degrading microorganisms. This paper explores the reactor conditions under which the polychlorinated biphenyl (PCB) degrader Burkholderia xenovorans LB400 can degrade significant amounts of the PCB mixture Aroclor(R) 1242. Aroclor(R) degradation was found to stall after approximately 40 h if no carbon source other than PCBs was available in the reactor. Sodium pyruvate was found to be a suitable carbon source to maintain microbial activity against PCBs and to function as a substrate for additional cell growth. Both biphenyl (while required during the inoculum preparation) and glucose had a negative effect during the Aroclor(R) degradation phase. Initial Aroclor(R) 1242 degradation rates in the presence of pyruvate were high (6.2 mg L(-1) h(-1)) and 85% of an equivalent concentration of 100 mg Aroclor(R) 1242 per L aqueous phase could be degraded in 48 h, which suggest that solvent extraction of PCBs from soil followed by their biodegradation in TPPBs might be a feasible remediation option.

Remediation of PAH contaminated soils: application of a solid-liquid two-phase partitioning bioreactor.

The feasibility of a two-step treatment process has been assessed at laboratory scale for the remediation of soil contaminated with a model mixture of polycyclic aromatic hydrocarbons (PAHs) (phenanthrene, pyrene, and fluoranthene). The initial step of the process involved contacting contaminated soil with thermoplastic, polymeric pellets (polyurethane). The ability of three different mobilizing agents (water, surfactant (Biosolve) and isopropyl alcohol) to enhance recovery of PAHs from soil was investigated and the results were compared to the recovery of PAHs from dry soil. The presence of isopropyl alcohol had the greatest impact on PAH recovery with approximately 80% of the original mass of PAHs in the soil being absorbed by the polymer pellets in 48 h. The second stage of the suggested treatment involved regeneration of the PAH loaded polymers via PAH biodegradation, which was carried out in a solid-liquid two-phase partitioning bioreactor. In addition to the PAH containing polymer pellets, the bioreactor contained a microbial consortium that was pre-selected for its ability to degrade the model PAHs and after a 14 d period approximately 78%, 62% and 36% of phenanthrene, pyrene, and fluoranthene, respectively, had been desorbed from the polymer and degraded. The rate of phenanthrene degradation was shown to be limited by mass transfer of phenanthrene from the polymer pellets. In case of pyrene and fluoranthene a combination of mass transfer and biodegradation rate might have been limiting.

Efficient Extraction of a Docosahexaenoic Acid (DHA)-Rich Lipid Fraction from Thraustochytrium sp. Using Ionic Liquids.

Polyunsaturated fatty acids (PUFAs) play a significant role in the modulation and prevention of various diseases, and hence are attracting increasing attention from the biotech industry. Thraustochytrids are marine heterokonts that exhibit robust growth rates, high PUFA content, and more specifically, a large percentage of omega-3 fatty acids like docosahexaenoic acid (DHA). Recently, ionic liquids (ILs) have been shown to improve the efficiency of organic solvent extraction of oils from wet oleaginous yeast and microalgae under mild conditions. Two ILs, the imidazolium 1-ethyl-3-methylimidazolium ethylsulfate [C2mim][EtSO4] IL and the phosphonium (tetrabutylphosphonium propanoate [P4444][Prop]) IL were assessed for their ability to facilitate extraction of PUFA-containing lipids from a Thraustochytrium sp. (T18) through efficient cell wall disruption. The oil extracted after IL pretreatment was further characterized with respect to fatty acid methyl ester (FAME) composition, while the effects of process parameters, such as the ratio of ionic liquid to co-solvent, the mass ratio of microalgae to the mixture of ionic liquid, and type of co-solvent were also investigated for both ILs. The results indicate that these ILs can disrupt the cells of Thraustochytrium sp. when mixed with a co-solvent (methanol), and facilitated the recovery of oils over a large degree of dewatered Thraustochytrium biomass (0⁻77.2 wt% water) in a short period of time (60 min) at ambient temperature, hence demonstrating a water compatible, low-energy, lipid recovery method. The lipid recovery was not affected by repeated usage of recycled ILs (tested up to five times).

Deep Eutectic Solvents pretreatment of agro-industrial food waste.

BACKGROUND: Waste biomass from agro-food industries are a reliable and readily exploitable resource. From the circular economy point of view, direct residues from these industries exploited for production of fuel/chemicals is a winning issue, because it reduces the environmental/cost impact and improves the eco-sustainability of productions. RESULTS: The present paper reports recent results of deep eutectic solvent (DES) pretreatment on a selected group of the agro-industrial food wastes (AFWs) produced in Europe. In particular, apple residues, potato peels, coffee silverskin, and brewer's spent grains were pretreated with two DESs, (choline chloride-glycerol and choline chloride-ethylene glycol) for fermentable sugar production. Pretreated biomass was enzymatic digested by commercial enzymes to produce fermentable sugars. Operating conditions of the DES pretreatment were changed in wide intervals. The solid to solvent ratio ranged between 1:8 and 1:32, and the temperature between 60 and 150 °C. The DES reaction time was set at 3 h. Optimal operating conditions were: 3 h pretreatment with choline chloride-glycerol at 1:16 biomass to solvent ratio and 115 °C. Moreover, to assess the expected European amount of fermentable sugars from the investigated AFWs, a market analysis was carried out. The overall sugar production was about 217 kt yr-1, whose main fraction was from the hydrolysis of BSGs pretreated with choline chloride-glycerol DES at the optimal conditions. CONCLUSIONS: The reported results boost deep investigation on lignocellulosic biomass using DES. This investigated new class of solvents is easy to prepare, biodegradable and cheaper than ionic liquid. Moreover, they reported good results in terms of sugars' release at mild operating conditions (time, temperature and pressure).