Pretreatment of wood by UCT-solvent for the enzymatic hydrolysis

UCT-solvent pretreatment was carried out on woods (beech and akamatsu (pine)) for the enzymatic hydrolysis, in which pretreatment the ground woods were autoclaved with a mixture of water and cyclo-hexanol (37.5% vol% cyclohexanol) having upper critical temperature (UCT: 184°C) on the mutual solubility curve (named as UCT-solvent). Ninety-five and 92% of Klason lignin were removed from beech and akamatsu, respectively, whereas when the woods were autoclaved with water instead of UCT-solvent, only 43 and 18% of Klason lignin was removed from them, respectively. The excellent ability of UCT-solvent for the removal of Klason lignin is owing to that the solvent disturbs re-coupling between the degradation products. The enzymatic hydrolysis of wood was much improved by UCT-solvent pretreatment: the hydrolytic reactivity of akamatsu was enhanced by 2.8 times comparing with when akamatsu was pretreated with water instead of UCT-solvent.     

Lime Pretreatment and Fermentation of Enzymatically Hydrolyzed Sugarcane Bagasse

Sugarcane bagasse was subjected to lime (calcium hydroxide) pretreatment and enzymatic hydrolysis for second-generation ethanol production. A central composite factorial design was performed to determine the best combination of pretreatment time, temperature, and lime loading, as well as to evaluate the influence of enzymatic loadings on hydrolysis conversion. The influence of increasing solids loading in the pretreatment and enzymatic hydrolysis stages was also determined. The hydrolysate was fermented using Saccharomyces cerevisiae in batch and continuous mode. In the continuous fermentation, the hydrolysates were concentrated with molasses. Lime pretreatment significantly increased the enzymatic digestibility of sugarcane bagasse without the need for prior particle size reduction. In the optimal pretreatment conditions (90 h, 90 °C, 0.47 g?lime/g bagasse) and industrially realistic conditions of hydrolysis (12.7 FPU/g of cellulase and 7.3 CBU/g of β-glucosidase), 139.6 kg?lignin/ton raw bagasse and 126.0 kg hemicellulose in the pretreatment liquor per ton raw bagasse were obtained. The hydrolysate from lime pretreated sugarcane bagasse presented low amounts of inhibitors, leading to ethanol yield of 164.1 kg?ethanol/ton raw bagasse.     

Enhancement of the Enzymatic Digestibility and Ethanol Production from Sugarcane Bagasse by Moderate Temperature-Dilute Ammonia Treatment

In this study, sugarcane bagasse was pretreated with ammonium hydroxide, and the effectiveness of the pretreatment on enzyme hydrolysis and ethanol production was examined. Bagasse was soaked in ammonium hydroxide and water at a ratio of 1:0.5:8 for 0–4 days at 70 °C. Approximately, 14–45 % lignin, 2–6 % cellulose, and 13–22 % hemicellulose were removed during a 0.5- to 4-day ammonia soaking period. The highest glucan conversion of sugarcane bagasse soaked in dilute ammonia at moderate temperature by cellulase was accomplished at 78 % with 75 % of the theoretical ethanol yield. Under the same conditions, untreated bagasse resulted in a cellulose digestibility of 29 and 27 % of the theoretical ethanol yield. The increased enzymatic digestibility and ethanol yields after dilute ammonia pretreatment was related to a combined effect of the removal of lignin and increase in the surface area of fibers.     

Kinetics of Lime Pretreatment of Sugarcane Bagasse to Enhance Enzymatic Hydrolysis

The objective of this work was to determine the optimum conditions of sugarcane bagasse pretreatment with lime to increase the enzymatic hydrolysis of the polysaccharide component and to study the delignification kinetics. The first stage was an evaluation of the influence of temperature, reaction time, and lime concentration in the pretreatment performance measured as glucose release after hydrolysis using a 2³ central composite design and response surface methodology. The maximum glucose yield was 228.45 mg/g raw biomass, corresponding to 409.9 mg/g raw biomass of total reducing sugars, with the pretreatment performed at 90°C, for 90 h, and with a lime loading of 0.4 g/g dry biomass. The enzymes loading was 5.0 FPU/dry pretreated biomass of cellulase and 1.0 CBU/dry pretreated biomass of β-glucosidase. Kinetic data of the pretreatment were evaluated at different temperatures (60°C, 70°C, 80°C, and 90°C), and a kinetic model for bagasse delignification with lime as a function of temperature was determined. Bagasse composition (cellulose, hemicellulose, and lignin) was measured, and the study has shown that 50% of the original material was solubilized, lignin and hemicellulose were selectively removed, but cellulose was not affected by lime pretreatment in mild temperatures (60–90°C). The delignification was highly dependent on temperature and duration of pretreatment.     

Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood

The wet oxidation pretreatment (water, oxygen, elevated temperature, and pressure) of softwood (Picea abies) was investigated for enhancing enzymatic hydrolysis. The pretreatment was preliminarily optimized. Six different combinations of reaction time, temperature, and pH were applied, and the compositions of solid and liquid fractions were analyzed. The solid fraction after wet oxidation contained 58–64% cellulose, 2–16% hemicellulose, and 24–30% lignin. The pretreatment series gave information about the roles of lignin and hemicellulose in the enzymatic hydrolysis. The temperature of the pretreatment, the residual hemicellulose content of the substrate, and the type of the commercial cellulase preparation used were the most important factors affecting the enzymatic hydrolysis. The highest sugar yield in a 72-h hydrolysis, 79% of theoretical, was obtained using a pretreatment of 200°C for 10 min at neutral pH.     

Pretreatment with ammonia water for enzymatic hydrolysis of corn husk, bagasse, and switchgrass

Bagasse, corn husk, and switchgrass were pretreated with ammonia water to enhance enzymatic hydrolysis. The sample (2 g) was mixed with 1–6 mL ammonia water (25–28% ammonia) and autoclaved at 120°C for 20 min. After treatment, the product was vacuum-dried to remove ammonia gas. The dried solid could be used immediately in the enzymatic hydrolysis without washing. The enzymatic hydrolysis was effectively improved with more than 0.5 and 1 mL ammonia water/g for corn husk and bagasse, respectively. In bagasse, glucose, xylose, and xylobiose were the main products. The adsorption of CMCase and xylanase was related to the initial rate of enzymatic hydrolysis. In corn husks, arabinoxylan extracted by pretreatment was substantially unhydrolyzed because of the high ratio of arabinose to xylose (0.6). The carbohydrate yields from cellulose and hemicellulose were 72.9% and 82.4% in bagasse, and 86.2% and 91.9% in corn husk, respectively. The ammonia/water pretreatment also benefited from switchgrass (Miscanthus sinensis and Solidago altissima L.) hydrolysis.     

Plasma-Assisted Pretreatment of Wheat Straw

O₃ generated in a plasma at atmospheric pressure and room temperature, fed with dried air (or oxygen-enriched dried air), has been used for the degradation of lignin in wheat straw to optimize the enzymatic hydrolysis and to get more fermentable sugars. A fixed bed reactor was used combined with a CO₂ detector and an online technique for O₃ measurement in the fed and exhaust gas allowing continuous measurement of the consumption of O₃. This rendered it possible for us to determine the progress of the pretreatment in real time (online analysis). The process time can be adjusted to produce wheat straw with desired lignin content because of the online analysis. The O₃ consumption of wheat straw and its polymeric components, i.e., cellulose, hemicellulose, and lignin, as well as a mixture of these, dry as well as with 50% water, were studied. Furthermore, the process parameters dry matter content and milled particle size (the extent to which the wheat straw was milled) were investigated and optimized. The developed methodology offered the advantage of a simple and relatively fast (0.5–2 h) pretreatment allowing a dry matter concentration of 45–60%. FTIR measurements did not suggest any structural effects on cellulose and hemicellulose by the O₃ treatment. The cost and the energy consumption for lignin degradation of 100 g of wheat straw were calculated.     

Impact of fluid velocity on hot water only pretreatment of corn stover in a flowthrough reactor

Flowthrough pretreatment with hot water only offers many promising features for advanced pretreatment of biomass, and a better understanding of the mechanisms responsible for flowthrough behavior could allow researchers to capitalize on key attributes while overcoming limitations. In this study, the effect of fluid velocity on the fate of total mass, hemicellulose, and lignin was evaluated for hot water only pretreatment of corn stover in tubular flow through reactors. Increasing fluid velocity significantly accelerated solubilization of total mass, hemicellulose, and lignin at early times. For example, when fluid velocity was increased from 2.8 to 10.7 cm/min, xylan removal increased from 60 to 82% for hot water only pretreatment of corn stover at 200°C after 8 min. At the same time, lignin removal increased from 30 to 46%. Dissolved hemicellulose was almost all in oligomeric form, and solubilization of hemicellulose was always accompanied by lignin release. The increase in removal of xylan and lignin with velocity, especially in the early reaction stage, suggests that chemical reaction is not the only factor controlling hemicellulose hydrolysis and that mass transfer and other physical effects may also play an important trole in hemicellulose and lignin degradation and removal.     

The effects of lignin removal and drying on the porosity and enzymatic hydrolysis of sugarcane bagasse

The porosity of lignocellulosic materials is a key feature for the enzymatic hydrolysis of the constituent polysaccharides, being affected by its drying and lignin content. Here we evaluated the influence of both parameters in the porosity of sugarcane bagasse, using raw and chlorite-delignified samples. A fraction of these samples was air dried at room temperature and the remainder one was kept wet. All the samples were subjected to porometry (solute exclusion technique), determination of water retention value and assessment of enzymatic saccharification of polysaccharides. Samples with higher lignin contents had lower porosities and exhibited worse enzymatic conversions of polysaccharides. Mild drying collapsed only the smaller pores, which already were inaccessible to enzymes, and therefore did not affect the efficiencies of saccharification. Our results show that the lignin content plays an important role in the porosity of the material and that its removal improves the enzymatic saccharification of the constituent polysaccharides.     

Utilization of waste cellulose

A pretreatment of lignocellulosic materials with sodium hypochlorite-hypochlorous acid at controlled pH (between 7 and 9) considerably increases the accessibility of the cellulosic part of the substrate to chemical and biochemical reactants. As a consequence, the yield and rate of the enzymatic hydrolysis to glucose is largely in creased. Wheat straw and spruce sawdust have been investigated. The increase in accessibility is assigned to degradation and (or) de tachment of the lignin network. The loss in cellulose and hemicellulose is not important, lignin being preferentially degraded under carefully controlled pH conditions. When applied to pure cel lulose, the pretreatment decreases the yield of enzymatic hydrolysis; in the absence of lignin, oxidation of the anhydroglucose units is im portant and results in the inhibition of the enzymatic hydrolysis.     

Effects of hemicellulose and lignin on enzymatic hydrolysis of cellulose from dairy manure

This study focused on the effect of hemicellulose and lignin on enzymatic hydrolysis of dairy manure and hydrolysis process optimization to improve sugar yield. It was found that hemicellulose and lignin in dairy manure, similar to their role in other lignocellulosic material, were major resistive factors to enzymatic hydrolysis and that the removal of either of them, or for best performance, both of them, improved the enzymatic hydrolysis of manure cellulose. This result combined with scanning electron microscope (SEM) pictures further proved that the accessibility of cellulose to cellulase was the most important feature to the hydrolysis. Quantitatively, fed-batch enzymatic hydrolysis of fiber without lignin and hemicellulose had a high glucose yield of 52% with respect to the glucose concentration of 17 g/L at a total enzyme loading of 1300 FPU/L and reaction time of 160 h, which was better than corresponding batch enzymatic hydrolysis.     

Optimization of Dilute Sulfuric Acid Pretreatment to Maximize Combined Sugar Yield from Sugarcane Bagasse for Ethanol Production

Increasing fermentable sugar yields per gram of biomass depends strongly on optimal selection of varieties and optimization of pretreatment conditions. In this study, dilute acid pretreatment of bagasse from six varieties of sugarcane was investigated in connection with enzymatic hydrolysis for maximum combined sugar yield (CSY). The CSY from the varieties were also compared with the results from industrial bagasse. The results revealed considerable differences in CSY between the varieties. Up to 22.7 % differences in CSY at the optimal conditions was observed. The combined sugar yield difference between the best performing variety and the industrial bagasse was 34.1 %. High ratio of carbohydrates to lignin and low ash content favored the release of sugar from the substrates. At mild pretreatment conditions, the differences in bioconversion efficiency between varieties were greater than at severe condition. This observation suggests that under less severe conditions the glucose recovery was largely determined by chemical composition of biomass. The results from this study support the possibility of increasing sugar yields or improving the conversion efficiency when pretreatment optimization is performed on varieties with improved properties.     

Lime Pretreatment of Sugarcane Bagasse for Bioethanol Production

The pretreatment of sugarcane bagasse with lime (calcium hydroxide) is evaluated. The effect of lime pretreatment on digestibility was studied through analyses using central composite design (response surface), considering pretreatment time, temperature, and lime loading as factors. The responses evaluated were the yield of glucose from pretreated bagasse after enzymatic hydrolysis. Experiments were performed using the bagasse as it comes from an alcohol/sugar factory (non-screened bagasse) and bagasse in the size range from 0.248 to 1.397 mm (screened bagasse) (12-60 mesh). It was observed that the particle size presented influence in the release of fermentable sugars after enzymatic hydrolysis using low loading of cellulase and β-glucosidase (3.5 FPU/g dry pretreated biomass and 1.0 IU/g dry pretreated biomass, respectively).     

Optimization of lignin recovery from sugarcane bagasse using ionic liquid aided pretreatment

Ionic liquid 1-ethyl-3-methylimidazolium acetate ([EMIM]oAc) was employed for the pretreatment of sugarcane bagasse (SCB) and extraction of lignin, a potentially valuable by-product of the biofuel industry. Response surface methodology based on central composite design was exploited and thereby an empirical model, exhibiting a coefficient of determination, R², of 0.9890, was established to optimize lignin recovery. In particular, a maximum lignin yield, equal to 90.1%, was calculated at the optimal pretreatment conditions, namely time: 120 min, temperature: 140 °C, and ionic liquid to bagasse ratio equal to 20:1 (wt/wt). The presence of guaiacyl and syringyl rings in lignin was confirmed by Fourier transform infrared spectroscopy (FTIR); whereas UV–Vis spectrophotometry showed that both p-coumaric acid and ferulic acid were contained in the lignin. Thermal analysis indicated a maximum decomposition rate of 2%/°C at 265 °C while Gel permeation chromatography analysis revealed that the molecular weight (M_w) of recovered lignin was equal to 1769 g/mol. Comparison of FTIR spectra of pretreated and untreated bagasse showed a negligible presence of lignin in the pretreated samples. Maximum delignification of bagasse after pretreatment was thus ensured. Thermal stability of the ionic liquid towards recyclability was proven by thermogravimetric analysis. The present study established adequate performance of neat and recycled ([EMIM]oAc) with regard to lignin recovery from SCB.     

Effects of Liquid Hot Water Pretreatment on Enzymatic Hydrolysis and Physicochemical Changes of Corncobs

Liquid hot water (LHW) pretreatment is an efficient chemical-free strategy for enhancing enzymatic digestibility of lignocellulosic biomass for conversion to fuels and chemicals in biorefinery. In this study, effects of LHW on removals of hemicelluloses and lignin from corncobs were studied under varying reaction conditions. LHW pretreatment at 160 °C for 10 min promoted the highest levels of hemicellulose solubilization into the liquid phase, resulting into the maximized pentose yield of 58.8% in the liquid and more than 60% removal of lignin from the solid, with 73.1% glucose recovery from enzymatic hydrolysis of the pretreated biomass using 10 FPU/g Celluclast?. This led to the maximal glucose and pentose recoveries of 81.9 and 71.2%, respectively, when combining sugars from the liquid phase from LHW and hydrolysis of the solid. Scanning electron microscopy revealed disruption of the intact biomass structure allowing increasing enzyme’s accessibility to the cellulose microfibers which showed higher crystallinity index compared to the native biomass as shown by x-ray diffraction with a marked increase in surface area as revealed by BET measurement. The work provides an insight into effects of LHW on modification of physicochemical properties of corncobs and an efficient approach for its processing in biorefinery industry.     

Two-Stage Fractionation of Corn Stover Using Aqueous Ammonia and Hot Water

Hot water and aqueous ammonia fractionation of corn stover were used to separate hemicellulose and lignin and improve enzymatic digestibility of cellulose. A two-stage approach was used: The first stage was designed to recover soluble lignin using aqueous ammonia at low temperature, while the second stage was designed to recover xylan using hot water at high temperature. Specifically, the first stage employed a batch reaction using 15 wt.% ammonia at 60 °C, in a 1:10 solid:liquid ratio for 8 h, while the second stage employed a percolation reaction using hot water, 190–210 °C, at a 20 ml/min flow rate for 10 min. After fractionation, the remaining solids were nearly pure cellulose. The two-stage fractionation process achieved 68% lignin purity with 47% lignin recovery in the first stage, and 78% xylan purity, with 65% xylan recovery in the second stage. Two-stage treatment enhanced the enzymatic hydrolysis of remaining cellulose to 96% with 15 FPU/g of glucan using commercial cellulase enzymes. Enzyme hydrolyses were nearly completed within 12–24 h with the remaining solids fraction.     

Compositional Changes in Sugarcane Bagasse on Low Temperature, Long-term Diluted Ammonia Treatment

Sugarcane bagasse is the major by-product of the sugar industry. It has a great potential for the production of biofuels and chemicals due to its considerable amount of cellulose and hemicellulose. In this study, we investigated a simple and economic pretreatment process using dilute ammonia for the storage of sugarcane bagasse. Sugarcane bagasse was stored in 0, 0.03, and 0.3% (w/w) ammonium hydroxide in a closed bottle for 40 days at 30 °C under atmospheric pressure without any agitation or circulation. Samples were taken every 10 days and analyzed for changes on lignin, cellulose, hemicellulose composition, ammonia concentration, and microbial counts. Biomass storage for 40 days at 0.3% ammonium hydroxide removed 46% of lignin and retained 100% cellulose and 73% hemicellulose.     

Evaluation and Characterization of Forage Sorghum as Feedstock for Fermentable Sugar Production

Sorghum is a tropical grass grown primarily in semiarid and drier parts of the world, especially areas too dry for corn. Sorghum production also leaves about 58 million tons of by-products composed mainly of cellulose, hemicellulose, and lignin. The low lignin content of some forage sorghums such as brown midrib makes them more digestible for ethanol production. Successful use of biomass for biofuel production depends on not only pretreatment methods and efficient processing conditions but also physical and chemical properties of the biomass. In this study, four varieties of forage sorghum (stems and leaves) were characterized and evaluated as feedstock for fermentable sugar production. Fourier transform infrared spectroscopy and X-ray diffraction were used to determine changes in structure and chemical composition of forage sorghum before and after pretreatment and the enzymatic hydrolysis process. Forage sorghums with a low syringyl/guaiacyl ratio in their lignin structure were easy to hydrolyze after pretreatment despite the initial lignin content. Enzymatic hydrolysis was also more effective for forage sorghums with a low crystallinity index and easily transformed crystalline cellulose to amorphous cellulose, despite initial cellulose content. Up to 72% hexose yield and 94% pentose yield were obtained using modified steam explosion with 2% sulfuric acid at 140 °C for 30 min and enzymatic hydrolysis with cellulase (15 filter per unit (FPU)/g cellulose) and β-glucosidase (50 cellobiose units (CBU)/g cellulose).     

Sugarcane Bagasse Mild Alkaline/Oxidative Pretreatment for Ethanol Production by Alkaline Recycle Process

In order to decrease the alkali and water consumptions in the sugarcane bagasse alkaline/oxidative pretreatment for ethanol production, an alkaline recycle process was carried out. Two recycles of NaOH/H₂O₂ pretreatment did not decrease the pretreatment and enzymatic hydrolysis efficiencies and the consumptions of NaOH and water would be saved by 26% and 40%, respectively. A simultaneous saccharification and fermentation (SSF) culture with pretreated bagasse as substrate was developed giving 25 g ethanol l⁻¹ with a yield of 0.2 g g⁻¹ bagasse and productivity of 0.52 g l⁻¹ h⁻¹.     

Pretreatment of sugarcane bagasse by acidified aqueous polyol solutions

Pretreatments of sugarcane bagasse by three high boiling-point polyol solutions were compared in acid-catalysed processes. Pretreatments by ethylene glycol (EG) and propylene glycol solutions containing 1.2 % H₂SO₄ and 10 % water at 130 °C for 30 min removed 89 % lignin from bagasse resulting in a glucan digestibility of 95 % with a cellulase loading of ~20 FPU/g glucan. Pretreatment by glycerol solution under the same conditions removed 57 % lignin with a glucan digestibility of 77 %. Further investigations with EG solutions showed that increases in acid content, pretreatment temperature and time, and decrease in water content improved pretreatment effectiveness. A good linear correlation of glucan digestibility with delignification was observed with R² = 0.984. Bagasse samples pretreated with EG solutions were characterised by scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray diffraction, which confirmed that improved glucan enzymatic digestibility is mainly due to delignification and defibrillation of bagasse. Pretreatment by acidified EG solutions likely led to the formation of EG-glycosides. Up to 36 % of the total lignin was recovered from pretreatment hydrolysate, which may improve the pretreatment efficiency of recycled EG solution.