Glucan Yield Research Articles

Ultrasound, microwave and enzyme-assisted multiproduct biorefinery of Ascophyllum nodosum

This study investigated the multiproduct (fucoidans, β-glucans, proteins, carotenoids, fatty acids, amino acids and polyphenols) valorization of the invasive macroalgae Ascophyllum nodosum within a green biorefinery concept using ultrasound (US), microwave (MW) treatment followed by supercritical CO2 (SC-CO2) with co-solvent and enzymatic extraction. Water and 50% aqueous ethanol were used as green extraction solvents. The extraction methods using 50% ethanol as extraction solvent improved the yields of phenolic compounds and glucan and enhanced in vitro antioxidant activity. The characterization of SC-CO2 extracts revealed that pretreatment with US and MW improved the 2-fold yield of carotenoids, total phenolics and fatty acids. However, US/MW pretreatment and enzymatic extraction did not improve the yields of proteins and free amino acids. Overall, using concurrent green US/MW-assisted extraction methods enhanced the yields of the bioactive compounds in a short duration and provided extracts with a better antioxidant capacity in the field of food applications.

Studies on metabolic regulation of Schizosaccharomyces pombe biomass production for glucan yield improvement

Studies on metabolic regulation of Schizosaccharomyces pombe biomass production for glucan yield improvement

Understanding triethylammonium hydrogen sulfate ([TEA][HSO4]) pretreatment induced changes in Pennisetum polystachion cell wall matrix and its implications on biofuel yield

Biofuel potential of a widely grown perennial grass, Pennisetum polystachion, was analyzed using triethylammonium hydrogen sulfate ([TEA][HSO4]) pretreatment. The optimum pretreatment condition was selected based on the delignification efficiency. 80% [TEA][HSO4] at 140 °C for 45 min at 10% sample load, yielding high delignification rate (65.8%) was selected as the optimum pretreatment condition. Recycling of [TEA][HSO4] showed up to 90% IL recovery and significant delignification rates. Glucan yield in the biomass increased to 67.8%, whereas digestibility of biomass enhanced from 16.7% to 84.1%. Extensive deferuloylation (88%) and decoumarylation (86.4%) were observed in the sample following pretreatment. Diferulic acids were not detected in pretreated samples indicating their complete removal. HMF (0.1 mg/mL) and furfural (0.001 mg/mL) produced during pretreatment were very low compared to conventional methods. FTIR and XRD confirmed pronounced delignification and hemicellulose dissolution, and FESEM showed a marked difference in the surface morphology, including defibrillation and enhanced porosity. An ethanol yield of 275 mg/g biomass (84.7% of theoretical maxima) was achieved using Saccharomyces cerevisiae MTCC 36 after 15 h of fermentation. The results obtained from the current study can shed light on utilizing P. polystachion as a potential biofuel feedstock using a low-cost ionic liquid [TEA][HSO4].

Investigation of extraction method effect on yeast beta glucan production

Nowadays, due to the stressful, tiring and busy lives of humans, the immune system becomes weak and can get sick easily. Therefore, scientists have been doing researches about new, natural and healthy products that can strengthen their immune system and can provide to adapt life standards. One of these products is the beta glucan. It is a polysaccharide molecule that consists of D-glucose monomers bonding with beta glycosidic bonds. Beta (β) glucans have being produced from different sources (microorganisms, cereals and mushrooms) so they have different branched structures such as (1→3) (1→6), (1→3) (1→4), (1→3) (1→2). Thus, different branched beta glucans show different physicochemical properties and biological activities that designate their usage purposes. Especially yeast beta glucan has lots of biological activities. On the other hand, it is fact that the extraction method affects the molecular weight, yield, purity and other properties of beta glucans. The main purpose of this study is to compare the performances of ultrasonically assisted alkali-acidic and autolysis extraction methods to produce a high yield of beta glucan. Also, it was aimed to determine the effect of extraction method on the molecular weight of yeast beta glucan. As a result of this work, it was found that the yeast beta glucan yield for ultrasonic supported autolysis extraction (12 %) is higher than that of the ultrasonic supported alkali-acidic extraction (8 %). On the other hand, having the lower molecular weight of yeast beta glucan (87 kDa) was produced by using an ultrasonic supported autolysis extraction method.

Green Husk of Walnuts (Juglans regia L.) from Southern Italy as a Valuable Source for the Recovery of Glucans and Pectins

Walnut green husk is an agricultural waste produced during the walnut (Juglans regia L.) harvest, that could be valued as a source of high-value compounds. In this respect, walnut green husks from two areas of Southern Italy (Montalto Uffugo and Zumpano), with different soil conditions, were investigated. Glucans and pectins were isolated from dry walnut husks by carrying out alkaline and acidic extractions, respectively, and then they were characterized by FT-IR, scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). The colorimetric method for the enzymatic measurement of α- and β-glucans was performed. The maximum total glucan yield was recovered from Montalto walnut husks (4.6 ± 0.2 g/100 g DM) with a β-glucan percentage (6.3 ± 0.4) higher than that calculated for Zumpano walnut husks (3.6 ± 0.5). Thermal analysis (DSC) confirmed the higher degree of crystallinity of glucans from Zumpano. The pectin content for Montalto husks was found to be 2.6 times that of Zumpano husks, and the esterification degree was more than 65%. The results suggested that J. regia L. green husks could be a source of glucans and pectins, whose content and morphological and thermal characteristics were influenced by different soil and climate conditions.

Effects of milling combined with extremely low-liquid ammonia (ELLA) pretreatment to enhance enzymatic hydrolysis of corn stover

Lignocellulosic biomass is one of the largest carbohydrate sources and has huge potential for biofuels production. However, the problem with lignocellulosic feedstock is that it has useful sugars locked in by lignin, hemicellulose, and cellulose. Some kind of pretreatment; therefore is needed to make carbohydrate accessible which later can be fermented to produce ethanol. The results from this research indicated that the yields of glucan (93%) and xylan (82.8%) were improved by using milling combined with ELLA pretreatment. The optimal enzymatic hydrolysis efficiencies were obtained under 10 min for ball milling time, pretreatment at 1 h, temperature at 150°C, S/L = 0.5 and ammonia loading at 0.25 g-NH3/g-biomass. This method reduced the pretreatment time and short milling time and thus has potential of reducing the energy consumption and promising the application in the large scale.

<p><strong>Organosolv process for deconstruction of highly recalcitrant cotton stalks</strong></p>

Burning crop residues in a cotton fields adds to environmental pollution and it demands a sustainable solution. As an alternative to burning, cotton biomass can be fractionated into sugars which would not only mitigate the pollution concern but also bring value addition. Here, we propose a process for deconstruction of the highly recalcitrant cotton stalk using a combination of organic and mineral acid fractionation. It includes treatment with formic acid (67 wt. %) at a solid to liquid ratio of 1:4 for 30 min at 130°C. Subsequent treatment with 2% (w/w) nitric acid and de-esterification with 0.5 % (w/v) sodium carbonate resulted in delignification of about 89.01%. The enzymatic hydrolysis using 25 FPU of Cellic Ctec3 at a biomass load of 5% (w/v) gave a glucan yield of 68% in 48 h. This is one of the few studies on deconstruction of a highly recalcitrant biomass with high lignin content.

Effects of sulfuric acid promoter on biopolymer-derived fractions from empty plam fruit bunch under solvothermal fractionation process

A single step sulfuric acid catalyzed (H2SO4) under the organosolv process was studied for fractionation of empty palm fruit branches. The use of H2SO4 as an acid catalyst showed higher efficiency of the removal of hemicellulose with an increase in lignin removal from native EPFB into organic phase and leave glucan in solid fraction. The optimal condition showed the maximum glucan yields of 2.45 g and the highest lignin yields of 1.86 g was at 180oC, with 15 min residence time. The addition of H2SO4 catalyzed enhanced hemicellulose and lignin removal. 0.05% H2SO4 led to improve glucan yields of 3.17 g with the lower degradation of glucan and pentose in the aqueous phase. Under this condition, a 1.28 g recovery of lignin was obtained from the organic phase. Physicochemical analysis revealed intact cellulose fibers with the decrease in crystallinity, while the hemicellulose was partially recovered as mono- and oligomeric sugar. High-purity organosolv lignin with < 2% sugar cross-contamination was obtained with no major structural modification according to Fourier-transform infrared spectroscopy. This work represents an alternative process for efficient fractionation of lignocellulosic biomass in biorefineries.

Screening of glucan and pectin contents in broad bean (Vicia faba L.) pods during maturation

This study was performed to investigate broad bean pods (BBPs), a food waste by-product material, for their glucan and pectin contents, at three different stages of maturation, in order to provide basic support for the functional uses of BBPs and for the development of a potential market for bioactive ingredients from residual sources. In the first instance, the isolation of glucans was performed by carrying out an alkaline extraction on the dry BBP flour, at each stage of maturation. The isolated glucans were dialyzed and the crude and soluble samples were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy analyses. Afterwards, the colorimetric method for the enzymatic measurement of glucans was used. This procedure involved partial acid hydrolysis combined with an enzyme complex for the quantitative determination of α- and β-glucans. The results showed that, regardless of the used method, the glucan content increased with maturation. The maximum total glucan yield was recovered from BBPs at third stage of maturation (45 mg/g). The kit-based procedure revealed the variability of the β-glucan amount with maturation whose percentage, compared to total glucans, increased from 47.6 at the first stage up to 61.9 at the third stage. On the contrary, the pectin content in BBPs decreased with maturation. The yields ranged from 8.7% at the third stage to 17.2% at the first stage of maturation. The degree of esterification was determined by the classical titrimetric method and confirmed by the instrumental FT-IR method. The samples showed an average titrimetric value of 43.4% and the typical fingerprint in IR spectra with an average value of 50.5%. The degree of methoxylation remained almost unchanged during maturation, regardless of the method used.

Effect of microwave-assisted ionic liquid/acidic ionic liquid pretreatment on the morphology, structure, and enhanced delignification of rice straw

In the present study, was investigated an environmentally friendly method for pretreating lignocellulosic rice straw (RS) by using 1-butyl-3-methylimidazolium chloride ([Bmim]Cl) as an ionic liquid (IL) and 1-butyl-3-methylimidazolium hydrogen sulfate ([Bmim]HSO4) as an acidic-IL (Acidic-IL) under microwave irradiation (microwave-[Bmim]Cl and microwave-[Bmim]HSO4). The conversion of lignocellulosic biomass into simple sugars requires both efficient pretreatment and hydrolysis enzymes to produce biofuels and specialty chemicals. Therefore, the applied [Bmim]Cl, [Bmim]HSO4, microwave-[Bmim]Cl, and microwave-[Bmim]HSO4 to improve hydrolysis yields. Structural analyses of the pretreated solids were performed to understand the synergistic effects of [Bmim]Cl, and [Bmim]HSO4 pretreatment under microwave irradiation (microwave-[Bmim]Cl and microwave-[Bmim]HSO4) on the efficiencies of enzymatic hydrolyses. The results of a chemical composition analysis of untreated and all pretreated RS samples by using the difference pretreatment methods showed that significant lignin removal was achieved using microwave-[Bmim]Cl (57.02 ± 1.24%), followed by [Bmim]Cl only (41.01 ± 2.67%), microwave-[Bmim]HSO4 (20.77 ± 1.79%), and [Bmim]HSO4-only (16.88 ± 1.14%). The highest glucan yield and xylan conversion achieved through the enzymatic saccharification of microwave-[Bmim]Cl-regenerated cellulose was consistent with the observations obtained from a structural analysis, which indicated a more disrupted, amorphous structure, with lowered crystallinity index (CrI) and lateral order index (LOI) of cellulose polymers. Thus results demonstrated that the pretreatment of lignocellulosic biomass with [Bmim]Cl under microwave irradiation has potential as an alternative method for pretreating lignocellulosic materials.

The scale-up cultivation of Candida utilis in waste potato juice water with glycerol affects biomass and \u03b2(1,3)/(1,6)-glucan characteristic and yield

New ideas on production of yeast origin β-glucan preparations for industrial application are attracting interest considering market development of that high-value functional polysaccharide. Sellecting an efficient yeast producer and designing culture conditions are a prerequisite for obtaining high yield of β-glucan. The aim of this study was to describe at the first time the influence of the mode of cultivation (shake-flasks and batch fermentation) and time of culture on characteristic and yield of biomass and β(1,3)/(1,6)-glucan preparations of Candida utilis ATCC 9950 after cultivation in medium based on waste potato juice water supplemented with 10% of glycerol. After shake-flask culture, the biomass was characterized by higher protein content (app. 26.5%) compared to 19% after batch fermentation while the cultivation on a biofermentor scale promoted polysaccharides biosynthesis. The highest output of purified β(1,3)/(1,6)-glucan preparation (5.3 gd.w./L), containing app. 85% of that polysaccharide, was found after 48 h cultivation in biofermentor. Batch fermentation promoted biosynthesis of alkali-insoluble β(1,3)/(1,6)-glucan fraction, decreasing the content of β(1,6)-glucan. The yield of β(1,3)/(1,6)-glucan synthesis was 0.063 (g/g glycerol), while the productivity of that polysaccharide reached 0.094 (g/L/h). Longer batch fermentation (72 h) resulted in reduction of production efficiency of β-glucan preparation under studied conditions. The results of the study provide a new efficient biotechnological solution to produce high-value β-glucan preparations of C. utilis origin based on valorization of agro-waste potato juice water with glycerol.

Persulfate oxidizing system for biomass pretreatment and process optimization

The functions of radicals in the pretreatment of lignocellulose remain a fascinating topic about which there are more questions than answers. The effect of persulfate-based radical species on lignin degradation and carbohydrate yield was investigated. Wheat straw was pretreated with three persulfate activation systems which generated OH•, SO42−•, and O2•−. Radical species were monitored using nitrobenzene, anisole, and hexachloroethane, respectively. Pretreatment variables affecting sugar yield were optimized by using the response surface method based on central composite design. Enzymatic hydrolysis and compositional analysis were performed to quantify changes in biomass composition, lignin removal, and glucan yield. Fourier transmission infrared spectroscopic and scanning electron microscopy were used to characterize the effect of pretreatment on biomass structure. The maximum sugar yields were 66.8%, 49.4%, and 41.7% in base, heat, and hydrogen peroxide activated persulfate, separately, while untreated material showed only 3.7% glucan yield. The radical analysis confirmed the existence of radicals in these three systems. The results from compositional analysis and Fourier transmission infrared spectroscopic provided proof that parts of lignin and hemicellulose were degraded after the treatment. Scanning electron microscopy indicted that radical pretreatment converted the smooth surface of wheat straw to that of scattered, uneven morphology. Base-activated persulfate, capable of producing sulfate radical, hydroxyl radical, and superoxide anion radical was demonstrated as an efficient radical system to treat biomass.

Conversion of Lignocellulosic Material Into Fermentable Sugars

Enzymatic hydrolysis process of lignocellulosic biomass materials is difficult because of inherent structural features of biomass, which represents barriers that prevent complete hydrolysis; therefore, pretreatment techniques are necessary to render biomass highly digestible in enzymatic hydrolysis process. In this research, (non?) oxidative short-term lime pretreatment of willow wood was used. A weight of 11.40 g of willow wood was mixed with an excess of calcium hydroxide (0.4 g Ca(OH)2/g raw biomass) and water loading (15 g/g raw biomass). Lime pretreatment was carried out for various periods of time including 1, 2, 3.5, 5 and 6 h, with temperatures at 100, 113, 130, 147 and 1600C, and oxygen pressures as oxidativeagent (6, 9, 13.5, 17.8, 21 bar absolute). The optimization of both pretreatment and enzymatic hydrolysis were depended on the maximum overall yields of glucan and xylan after two processes of lime pretreatment and enzymatic hydrolysis. The optimal conditions of pretreatment were as follow: 1) 1.33 h, 1470C, 17.8 bar absolute, 0.26 g Ca(OH)2/g raw biomass. 2) 1.25 h, 155 0C, 21 bar absolute, 0.26 Ca(OH)2/g raw biomass. Furthermore, the optimal values for low impact factors such as water loading was 15 g/g raw biomass and particle size was less than 3 mm. The optimal conditions of enzymatic hydrolysis were as follow: Cellulase enzymeloading was 0.1 g /g glucan in raw biomass, at substrate concentration of 50 g/L during 72 h of enzymatic hydrolysis The yield of enzymatic hydrolysis under these conditions were as follow: 96.00 g glucan/100 g of glucan in raw biomass, and 65.00 g xylan/100 g xylan in raw biomass.

Improvement of radio frequency (RF) heating-assisted alkaline pretreatment on four categories of lignocellulosic biomass.

Pretreatment plays an important role in making the cellulose accessible for enzyme hydrolysis and subsequent conversion because it destroys more or less resistance and recalcitrance of biomass. Radio frequency (RF)-assisted dielectric heating was utilized in the alkaline pretreatment on agricultural residues (corn stover), herbaceous crops (switchgrass), hardwood (sweetgum) and softwood (loblolly pine). Pretreatment was performed at 90°C with either RF or traditional water bath (WB) heating for 1h after overnight soaking in NaOH solution (0.2g NaOH/g Biomass). Pretreated materials were characterized by chemical compositional analysis, enzyme hydrolysis, scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The glucan yields of RF-heated four categories of hydrolysates were 89.6, 72.6, 21.7, and 9.9%. Interestingly, RF heating raised glucan yield on switchgrass and sweetgum but not on corn stover or loblolly pine. The SEM images and FTIR spectra agreed with results of composition analysis and hydrolysis. GC-MS detected some compounds only from RF-heated switchgrass. These compounds were found by other researchers only in high-temperature (150-600°C) and high-pressure pyrolysis processes.

Natural genetic variability reduces recalcitrance in poplar

BackgroundLignin content and structure are known to affect recalcitrance of lignocellulosic biomass to chemical/biochemical conversion. Previously, we identified rare Populus trichocarpa natural variants with significantly reduced lignin content. Because reduced lignin content may lower recalcitrance, 18 rare variants along with 4 comparators, and BESC standard Populus was analyzed for composition of structural carbohydrates and lignin. Sugar yields from these plants were measured at 5 process conditions: one for just enzymatic hydrolysis without pretreatment and four via our combined high-throughput hot water pretreatment and co-hydrolysis (HTPH) technique.ResultsMean of glucan + xylan yields and the best glucan + xylan yield from rare natural poplar variants were 34 and 50 relative percent higher than the high lignin comparator (BESC-316) at the highest severity HTPH condition, respectively. The ability of HTPH to solubilize a large portion of xylan from solids led to small differences in xylan yields among poplar variants. However, HTPH showed large differences in glucan yields, and hence glucan + xylan yields, among the poplar variants. The high lignin comparator did not display lowest glucan + xylan yields with HTPH at moderate pretreatment severity compared to rare variants, but on the other hand, the low lignin comparator was a consistent top performer at all 5 process conditions. Furthermore, the low lignin comparator (GW-11012) showed a 15 absolute percent increase in glucan + xylan yield compared to the high lignin comparator at the most severe HTPH condition. Overall, relative variant rankings varied greatly with pretreatment severity, but poplar deconstruction was significantly enhanced when the pretreatment temperature was increased from 140 and 160 to 180 °C at the same pretreatment severity factor.ConclusionsGlucan yields from high severity HTPH of rare natural poplar variants with reduced lignin content were significantly higher than from the high lignin comparator. Because of the significant effect of processing conditions on the performance rankings, selection of the best performing biofuel feedstocks should be based on sugar yields tested at conditions that represent industrial practice. From a feedstock perspective, the most consistent variants, SKWE-24-2 and GW-11012, provide key insights into the genetic improvement of versatile lignocellulosic biofuels feedstock varieties.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0521-2) contains supplementary material, which is available to authorized users.

Ameliorated enzymatic saccharification of corn stover with a novel modified alkali pretreatment

Enzymatic saccharification/hydrolysis is one of the key steps for the bioconversion of lignocelluloses into sustainable biofuels. In this work, corn stover was pretreated with a novel modified alkali process (NaOH + anthraquinone (AQ) + sodium lignosulfonate (SLS)), and then enzymatically hydrolyzed with an enzyme cocktail (cellulase (Celluclast 1.5L), β-glucosidase (Novozyme 188) and xylanase (from thermomyceslanuginosus)) in the pH range of 4.0-6.5. It was found that the suitable pH for the enzymatic saccharification process to achieve a high glucan yield was between 4.2 and 5.7, while the appropriate pH to obtain a high xylan yield was in the range of 4.0-4.7. The best pH for the enzymatic saccharification process was found to be 4.4 in terms of the final total sugar yield, as xylanase worked most efficiently in the pH range of 4.0-4.7, under the conditions in the study. The addition of xylanase in the enzymatic saccharification process could hydrolyze xylan in the substrates and reduce the nonspecific binding of cellulase, thus improving the total sugar yields.

Reutilization of green liquor chemicals for pretreatment of whole rice waste biomass and its application to 2,3-butanediol production

The performance of green liquor pretreatment using Na2CO3 and Na2SO3 and its optimization for whole rice waste biomass (RWB) was investigated. Incubation of Na2CO3–Na2SO3 at a 1:1 ratio (chemical charge 10%) for 12% RWB at 100°C for 6h resulted in maximum delignification (58.2%) with significant glucan yield (88%) and total sugar recovery (545mg/g of RWB) after enzymatic hydrolysis. Recovery and reusability of the resulting chemical spent wash were evaluated to treat RWB along with its compatibility for enzymatic digestibility. Significant hydrolysis and lignin removal were observed for up to three cycles; however, further reuse of Na2CO3 and Na2SO3 lowered their performance. Significant 2,3-butanediol (BDO) was produced by Klebsiella pneumoniae KMK-05 with the RWB enzymatic hydrolysate from each pretreatment cycle. BDO yield achieved using RWB-derived sugars was similar to those using laboratory-grade sugars. This pretreatment strategy constitutes an ecofriendly, cost-effective, and practical method for utilizing lignocellulosic biomass.

Fractionation of rice straw by a single-step solvothermal process: Effects of solvents, acid promoters, and microwave treatment

Fractionation of lignocellulosic components is a primary step for maximizing value-added conversion of plant biomass to biofuels and chemicals. In this study, different organic solvents (methyl isobutyl keone, ethyl acetate, toluene and diethyl ether) in the ternary water/alcohol/organic solvent system and acid promoters (H2SO4, HCl, and H3PO4) were studied on a single-step solvothermal fractionation of rice straw. Ethyl acetate was found to be a superior composite solvent to the conventional solvent MIBK by providing higher glucan and lignin yields. Glucan and xylan could be further solubilized and lignin recovery increased at higher temperature and acid concentration. The optimal fractionation conditions were found to be water/ethanol/ethyl acetate (62.5%:25%:12.5%) mixture using 0.05 M H2SO4 at 160 °C for 1 h, which gave cellulose-enriched solid with 71.4 wt% glucan yield. Hemicellulose (53.2 wt%) was obtained as sugars, and dehydration products in the aqueous/alcohol phase were minimal. 63.1 wt% lignin was recovered in the organic solvent phase with no cross-contamination of sugars. Microwave-assisted heating at 300 W, corresponded to the temperature of 160 °C for 1 h resulted in comparable glucan and lignin yields in the respective phases to conventional heating with the advantages of increased pentose yield and lower furan formation in the aqueous phase.

Sugarcane hybrids with original low lignin contents and high field productivity are useful to reach high glucose yields from bagasse

Five sugarcane hybrids plus a reference material were evaluated according to the glucose yields obtained after alkaline-sulfite pretreatment and enzymatic hydrolysis. Sugarcane hybrids with varied original chemical compositions were used to assess how contrasting samples might influence the integrated pretreatment and hydrolysis process. The hydrolysis efficiency of six samples treated at three different chemical loads, suggested that lignin and hemicellulose removals during the pretreatment were not the single factor necessary to reach high cellulose conversion levels in the enzymatic hydrolysis step. Pretreated samples with the highest total acid contents (mainly sulfonic acids) were also the most digestible materials. The glucose yields were heavily dependent not only on the digestibility of the pretreated materials but also on the field productivity of the plants. One of the hybrids, presenting high glucan yields after pretreatment and high digestibility, produced low glucose yields because it presented very low biomass productivity. In contrast, one of the hybrids that presented low glucan yield after pretreatment, but was highly digestible and presented high biomass productivity, provided the highest glucose yields in the data set, producing 4192 and 5629 kg of glucose per hectare after enzymatic hydrolysis for 24 h and 72 h, respectively.

Scale-Up of Ionic Liquid-Based Fractionation of Single and Mixed Feedstocks

Lignocellulosic biorefineries have tonnage and throughput requirements that must be met year round, and there is no single feedstock available in any given region that is capable of meeting the price and availability demands of the biorefineries. Ionic liquid (IL) pretreatment with certain ILs is receiving significant attentions as a potential process that enables fractionation of a wide range of feedstocks and produces high yields of fermentable sugars suitable for biofuel production. Building on the large-scale demonstration of a single herbaceous feedstock (switchgrass), this work extends scale-up of IL pretreatment to woody (eucalyptus) and mixed feedstock (mixtures of two) by 30-fold, relative to the bench scale (6 vs 0.2 L) at 10 % solid loading. The mixed feedstock recovered similar yields of glucan (99.7 %), xylan (62.8 %), and lignin (59.9 %) as switchgrass and eucalyptus at 6-L scale operation, and results of all three feedstocks are better than those obtained from small-scale studies. By integrating the process of IL pretreatment with efficient and scalable homogenization, washing, and product recovery system, IL contents in the recovered materials were decreased to 0.2 %, mitigating the risk to downstream enzymatic saccharification and microbial fermentation. Results indicate that mixed feedstock are viable and valuable resource to consider when assessing biomass availability and affordability for lignocellulosic biorefineries. This scale-up evaluation demonstrates that IL pretreatment technology is feedstock agnostic and can be effectively scaled to larger operations.