Enzymatic Digestibility Of Cellulose Research Articles

Efficient fractionation of corn stover using deep eutectic solvent from lignin-derived 3-phenylpropionic acid: Towards a closed-loop biorefinery

The search for renewable deep eutectic solvents (DESs) for efficient biomass pretreatment based on a closed-loop biorefinery is of great interest and importance. Herein, we reported three novel lignin-based DESs prepared by choline chloride (ChCl) and 3-(4-hydroxyphenyl)propionic acid (HPA), 3-(4-hydroxy-3-methoxyphenyl)propanoic acid (HMPA) and their combination for corn stover pretreatment. The effects of three lignin-based DESs pretreatments on the chemical compositions of raw material, structural features of lignin, and enzymatic digestibility of cellulose were comprehensively analyzed, among which ChCl/HPA DES exhibited superior pretreatment efficiency, affording 58% of delignification, 62% of hemicellulose removal, and 85% of glucose yield. A high abundance of p-coumaric acid in the regenerated and remaining lignin samples makes them a sustainable feedstock to produce hydrogen bond donors of ChCl/HPA DES, as demonstrated by the results from 2D HSQC NMR analysis and catalytic depolymerization of lignins. These results could be conducive to creating an integrated biorefinery strategy for biomass valorization. Mass balance analysis indicated 81% of cellulose, 90% of hemicellulose, and 91% of lignin components in corn stover could be collected after DES pretreatment. This contribution offers a novel and efficient pretreatment method for corn stover fractionation based on a closed-loop biorefinery.

Insight into key obstacles and technological strategy for enzymatic digestion of full cellulose fraction from poplar sawdust

Lignocellulosic biomass mainly consists of hemicellulose, lignin, and cellulose, which differently affect the enzymatic digestibility of cellulose. As for the typical representative for inert woody biomass, three components of cellulose were proposed conceptually for poplar sawdust, i.e., active cellulose, inert cellulose, and resistant cellulose. Dilute sulfuric acid pretreatment, hydrogen peroxide-sulfuric acid delignification, and sulfuric acid-assisted glycerol swelling were, respectively, proven to break the three obstacle mechanisms that affect the cellulase of poplar. The removal of key obstacles improved the cellulase digestibility of poplar enzyme-hydrolyzed residues by 188.7 %, and glucose yield increased from 34.6 % to 99.9 %. Therefore, a total of 39.5 g glucose was obtained from 100 g poplar sawdust by integrating the above three technologies. This work presented insight into and removed the key obstacles to enzymatic digestibility of poplar cellulose and developed an integrated technology to effectively convert full cellulose fraction to glucose from woody biomass.

Novel supramolecular deep eutectic solvent-enabled in-situ lignin protection for full valorization of all components of wheat straw

Lignin is usually deemed as an inhibitor to enzymatic hydrolysis of cellulose due to its physical barrier, non-productive adsorption, and steric hindrance. Herein, a novel supramolecular deep eutectic solvent (SUPRADES), comprising ethylene glycol and citric acid in 5:1 M ratio, and β-cyclodextrin (β-CD) in a concentration of 3.5% (w/w), was developed to be efficient for pretreating wheat straw. The delignification rate, cellulose enzymatic digestibility, and hemicellulose removal reached 90.45%, 97.36% and 87.24%, respectively, which may be attributed to the introduction of β-CD with superior ability of both adsorption and in-situ lignin protection to efficiently remove lignin with intact structure from cellulose surface. The mechanisms of high-efficiency lignin extraction/protection were systematically illustrated by adsorption kinetics. Moreover, Trichosporon cutaneum grown on the hemicellulose and cellulose fractions after pretreatment afforded 8.8 g total lipids from 100 g wheat straw. The green SUPARDES pretreatment strategy offers a new avenue for upgrading lignocellulose to biofuels.

Glycolic acid-based deep eutectic solvents boosting co-production of xylo-oligomers and fermentable sugars from corncob and the related kinetic mechanism

BackgroundXylo-oligomers are a kind of high value-added products in biomass fractionation. Although there are several chemical methods to obtain xylo-oligomers from biomass, the reports about the deep eutectic solvents (DESs)-mediated co-production of xylo-oligomers and fermentable sugars and the related kinetic mechanism are limited.ResultsIn this work, glycolic acid-based DESs were used to obtain xylo-oligomers from corncob. The highest xylo-oligomers yield of 65.9% was achieved at 120 °C for 20 min, of which the functional xylo-oligosaccharides (XOSs, DP 2–5) accounted for up to 31.8%. Meanwhile, the enzymatic digestion of cellulose and xylan in residues reached 81.0% and 95.5%, respectively. Moreover, the addition of metal inorganic salts significantly accelerated the hydrolysis of xylan and even the degradation of xylo-oligomers in DES, thus resulting in higher selectivity of xylan removal. AlCl3 showed the strongest synergistic effect with DES on accelerating the processes, while FeCl2 is best one for xylo-oligomers accumulation, affording the highest xylo-oligomers yield of 66.1% for only 10 min. Furthermore, the kinetic study indicates that the ‘potential hydrolysis degree’ model could well describe the xylan hydrolysis processes and glycolic acid/lactic acid (3:1) is a promising solvent for xylo-oligomers production, in particular, it worked well with FeCl2 for the excellent accumulation of xylo-oligomers.ConclusionsGlycolic acid-based deep eutectic solvents can be successfully applied in corncob fractionation with excellent xylo-oligomers and fermentable sugars yields on mild conditions, and the large amount of xylo-oligosaccharides accumulation could be achieved by specific process controlling. The strategies established here can be useful for developing high-valued products from biomass.

Evaluation of the Extraction of Bioactive Compounds and the Saccharification of Cellulose as a Route for the Valorization of Spent Mushroom Substrate.

The extraction of bioactive compounds and cellulose saccharification are potential directions for the valorization of spent mushroom substrate (SMS). Therefore, investigating the suitability of different extraction methods for recovering bioactive compounds from SMS and how the extraction affects the enzymatic saccharification is of uppermost relevance. In this work, bioactive compounds were extracted from Pleurotus spp. SMS using four extraction methods. For Soxhlet extraction (SoE), a 40:60 ethanol/water mixture gave the highest extraction efficiency (EE) (69.9-71.1%) among the seven solvent systems assayed. Reflux extraction with 40:60 ethanol/water increased the extraction yield and EE compared to SoE. A shorter reflux time yielded a higher extraction of carbohydrates than SoE, while a longer time was more effective for extracting phenolics. The extracts from 240 min of reflux had comparable antioxidant activity (0.3-0.5 mM GAE) with that achieved for SoE. Ultrasound-assisted extraction (UAE) at 65 °C for 60 min allowed an EE (~82%) higher than that achieved by either reflux for up to 150 min or SoE. Subcritical water extraction (SWE) at 150 °C resulted in the best extraction parameters among all the tested methods. Vanillic acid and chlorogenic acid were the primary phenolic acids identified in the extracts. A good correlation between the concentration of caffeic acid and the antioxidant activity of the extracts was found. Saccharification tests revealed an enhancement of the enzymatic digestibility of SMS cellulose after the extraction of bioactive compounds. The findings of this initial study provide indications on new research directions for maximizing the recovery of bioactive compounds and fermentable sugars from SMS.

Improved enzymatic saccharification of bulrush via an efficient combination pretreatment

Glycerol (Gly) was selected as hydrogen-bond-donor for preparing ChCl-based DES (ChCl:Gly), and the mixture of ChCl:Gly (20 wt%) and NaOH (4 wt%) was utilized for combination pretreatment of bulrush at 100 °C for 60 min (severity factor LogRo = 1.78). The effects of DES pretreatment on the chemical composition, microstructure, crystal structure, and cellulase hydrolysis were explored. NaOH-ChCl:Gly could remove lignin (80.1%) and xylan (66.8%), and the enzymatic digestibility of cellulose reached 87.9%. The accessibility of bulrush was apparently increased to 645.2 mg/g after NaOH-ChCl:Gly pretreatment. The hydrophobicity and lignin surface area were reduced to 1.56 L/g and 417 m2/g, respectively. The crystallinity of cellulose was increased from 20.8% to 55.6%, and great changes in surface morphology were observed, which explained the improvement of enzymatic hydrolysis efficiency. Overall, DES combined with alkali treatment could effectively promote the removal of lignin and xylan in bulrush, thus the relative saccharification activity was greatly affected.

Improving the Cellulose Enzymatic Digestibility of Sugarcane Bagasse by Atmospheric Acetic Acid Pretreatment and Peracetic Acid Post-Treatment.

Pretreatment of sugarcane bagasse (SCB) by aqueous acetic acid (AA), with the addition of sulfuric acid (SA) as a catalyst under mild condition (<110 °C), was investigated. A response surface methodology (central composite design) was employed to study the effects of temperature, AA concentration, time, and SA concentration, as well as their interactive effects, on several response variables. Kinetic modeling was further investigated for AA pretreatment using both Saeman's model and the Potential Degree of Reaction (PDR) model. It was found that Saeman's model showed a great deviation from the experimental results, while the PDR model fitted the experimental data very well, with determination coefficients of 0.95-0.99. However, poor enzymatic digestibility of the AA-pretreated substrates was observed, mainly due to the relatively low degree of delignification and acetylation of cellulose. Post-treatment of the pretreated cellulosic solid well improved the cellulose digestibly by further selectively removing 50-60% of the residual linin and acetyl group. The enzymatic polysaccharide conversion increased from <30% for AA-pretreatment to about 70% for PAA post-treatment.

Efficient delignification of wheat straw for microbial lipid production enabled by a novel ternary deep eutectic solvent containing ethylene glycol

Lignocellulose pretreatment often faces the problem of lignin condensation with less attention to biofuel production from total sugars, which significantly hinders biomass fractionation and energy utilization efficiency. The aim of this work was to design a novel ternary deep eutectic solvent (DES) (citric acid: choline chloride: ethylene glycol in a molar ratio of 1:1:2) for pretreatment of wheat straw with a solid/liquid ratio (S/L) of 1:20 (w/w) to achieve non-condensed lignin and high-yield fermentable sugars at 100 °C for 12 h. The introduction of ethylene glycol into ternary DES could capture the formed benzyl carbocation to protect lignin structure, inhibit excessive degradation of hemicellulose but to fermentable sugars (e.g., xylose), and improve the enzymatic digestibility of cellulose (98.73%, w/w). The mechanisms of both lignin fractionation and the in-situ lignin protection during the pretreatment of wheat straw enabled by ternary DES were systematically elucidated. Importantly, Trichosporon cutaneum cultivated in undetoxified DES waste liquid (hemicellulose-rich) and residue hydrolysate (cellulose-rich) could totally afford 8.7 g microbial lipid per 100 g wheat straw, which is superior to previous reports. This green in-situ lignin protection and fractionation strategy provides a new avenue for upgrading lignocellulose to microbial lipids.

Comparative study of acid- and alkali-catalyzed 1,4-butanediol pretreatment for co-production of fermentable sugars and value-added lignin compounds

BackgroundOrganosolv pretreatment is one of the most efficient methods for delignification and boosting biomass saccharification. As compared to typical ethanol organosolv pretreatments, 1,4-butanediol (BDO) organosolv pretreatment is a high-boiling-point solvent pretreatment, which can generate low pressure in the reactor during high temperature cooking that improves the operation safety. Although several studies showed that organosolv pretreatment can lead to effective delignification and enhancement in glucan hydrolysis, there has been no studies on acid- and alkali-catalyzed BDO pretreatment, as well as their comparison on promoting biomass saccharification and lignin utilization.ResultsIt was shown that BDO organosolv pretreatment was more effective in removing lignin from poplar as compared with typical ethanol organosolv pretreatment under the same pretreatment conditions. HCl-BDO pretreatment with 40 mM acid loading led to 82.04% of original lignin removed from biomass, as compared to the lignin removal of 59.66% in HCl-Ethanol pretreatment. Besides, acid-catalyzed BDO pretreatment was more effective in improving the enzymatic digestibility of poplar than alkali-catalyzed BDO pretreatment. As a result, HCl-BDO with acid loading of 40 mM provided a good enzymatic digestibility of cellulose (91.16%) and the maximum sugar yield of 79.41% from original woody biomass. The linear correlations between physicochemical structure (e.g., fiber swelling, cellulose crystallinity, crystallite size, surface lignin coverage and cellulose accessibility) changes of BDO pretreated poplar and enzymatic hydrolysis were plotted to figure out the main factors that influenced biomass saccharification. Moreover, acid-catalyzed BDO pretreatment mainly brought about the phenolic hydroxyl (PhOH) groups formation in lignin structure, while alkali-catalyzed BDO pretreatment mostly led to the lower molecular weight of lignin.ConclusionsResults indicated that the acid-catalyzed BDO organosolv pretreatment could significantly improve enzymatic digestibility of the highly recalcitrant woody biomass. The great enzymatic hydrolysis of glucan resulted from increased cellulose accessibility, which mostly associated with the higher degree of delignification and hemicellulose solubilization, as well as the more increase in fiber swelling. Besides, lignin was recovered from the organic solvent, which could be used as natural antioxidants. The formation of phenolic hydroxyl groups in lignin structure and the lower molecular weight of lignin contributed to its greater radical scavenging capacity.

The crystalline structure transition and hydrogen bonds shift determining enhanced enzymatic digestibility of cellulose treated by ultrasonication

Global energy issue raised the necessity to develop second-generation biofuels, and biorefinery of cellulosic biomass becomes a promising solution. Various pretreatments were used to overcome the cellulose nature of recalcitrance and improve the enzymatic digestibility, but the lack of mechanism understanding hindered the development of efficient and cost-effective technologies of cellulose utilization. Using structure-based analysis, we demonstrate that the improved hydrolysis efficiency caused by ultrasonication was ascribed to the changed cellulose properties rather than the increased dissolubility. Further, isothermal titration calorimetry (ITC) analysis suggested that enzymatic digestion of cellulose is an entropically favored reaction driven by hydrophobic forces other than an enthalpically favored reaction. The changes in cellulose properties and thermodynamic paramenters due to ultrasonication accounted for the improved accessibility. Ultrasonication-treated cellulose showed porous, rough and disordered morphology, accompanying with the loss of crystalline structure. Despite the unaffected unit cell structure, ultrasonication expanded the crystalline lattice by increasing grain sizes and average cross-sectional area, resulting in the transformation from cellulose I to cellulose II, with the decreased crystallinity, better hydrophilicity and increased enzymatic bioaccessibility. Furthermore, FTIR combined with two-dimensional correlation spectroscopy (2D-CoS) verified that the sequential shift of hydroxyl group and intramolecular/intermolecular hydrogen bonds, the functional groups governing cellulose crystal structure and stability, accounted for the ultrasonication-induced transition of cellulose crystalline structure. This study provides a comprehensive picture of cellulose structure and property response caused by mechanistic treatments and will open up avenues to develop novel pretreatments for efficient utilization.

Utilization of walnut shell by deep eutectic solvents: enzymatic digestion of cellulose and preparation of lignin nanoparticles

Utilization of walnut shell by deep eutectic solvents: enzymatic digestion of cellulose and preparation of lignin nanoparticles

A novel ternary deep eutectic solvent pretreatment for the efficient separation and conversion of high-quality gutta-percha, value-added lignin and monosaccharide from Eucommia ulmoides seed shells

A novel ternary deep eutectic solvent (DES), consisted of choline chloride, oxalic acid and ethylene glycol, was developed as a green, low-cost and recyclable pretreatment system for multi-stage utilization of Eucommia ulmoides seed shells. Under optimum conditions, 79.7 % hemicellulose and 65.6 % lignin were quickly removed while 84.0 % cellulose was retained. After DES pretreatment, the yield and purity of gutta-percha achieved 85.1 mg/g and 96.2 %, which increased 1.4 and 1.8 folds higher than that of un-treatment ones. Meanwhile, 69.1 % enzymatic digestibility of cellulose was obtained, that was 2.3 folds higher than that of raw substrates. Moreover, 53.6 % low-condensation lignin with aromatic structures and valuable aryl-ether linkages was well collected. Importantly, the DES that has been recycled five runs can still remove 73.9 % hemicellulose and 58.0 % lignin. Overall, the DES was determined to efficiently promote the separation and conversion of high-quality gutta-percha, value-added lignin and high-yield glucose from Eucommia ulmoides seed shells.

Elucidating the inhibiting mechanism of pseudo-lignin on the enzymatic digestibility of cellulose by surface plasmon resonance

To explore the interaction mechanism of pseudo-lignin (PL) with cellulase and its influence on cellulose hydrolysis, different PLs were extracted from pretreated bamboo holocellulose (HC) using different organic solvents. Meanwhile, the real-time interaction of PL and cellulase was analyzed using surface plasmon resonance (SPR). The results showed that the extraction effect of the tetrahydrofuran and 1, 4-dioxane/water solution on PL was more effective than the ethanol/water solution. The inhibition of PL fraction obtained from HC by acid pretreatment with higher temperature showed less effect on Avicel’s enzymatic hydrolysis. SPR analysis revealed that PL formed at higher pretreatment temperature had a lower dissociation rate after adsorption with cellulase. Besides, the binding affinity of PL (160 °C) to cellulase was much greater than that of PL obtained from 180 °C, indicating PL extracted at higher temperature treated biomass is more easily dissociated from cellulase after binding.

Microstructural and histochemical variations of Korshinsk peashrub during aqueous ammonia pretreatment and its impacts on enzymatic digestibility of cellulose

Korshinsk peashrub (KP) is a drought-tolerant perennial energy shrub. Valorization of KP is beneficial to both environment protection and economy development. Herein, aqueous ammonia (AA) pretreatment was applied to enhance enzymatic hydrolysis of KP for the first time, and influences of AA pretreatment on recalcitrance of KP were also assessed through wet chemistry composition, physical properties, and glycome profiling analysis. The results suggested that AA pretreatment at 80 ◦C degraded over 9.85% neutral detergent soluble and retained 51.63% cellulose. AA pretreatment could explicitly eliminate non-cellulosic polymers, e.g. pectin/arabinogalactan epitopes, and interrupt ester bond and pectin linking matrix, leading to deconstruction in overall cell wall integrity. The pretreated KP (80 ◦C) gave the highest reducing sugar yield of 316.93 mg/g, which was 1.96 times as high as the untreated KP. AA pretreatment at atmospheric temperature might be a promising strategy employed in enzymatic saccharification of KP in biorefinery.

A hydrotrope pretreatment for stabilized lignin extraction and high titer ethanol production

The biomass pretreatment strategies using organic acids facilitate lignin removal and enhance the enzymatic digestion of cellulose. However, lignin always suffers a severe and irreversible condensation. The newly generated C–C bonds dramatically affect its further upgrading. In this study, we used a recyclable hydrotrope (p-Toluenessulfonic acid, p-TsOH) to dissolve lignin under mild condition and stabilized lignin with a quenching agent (formaldehyde, FA) during extraction, achieving both value-added lignin extraction and efficient enzymatic saccharification of cellulose. Approximately 63.7% of lignin was dissolved by 80% (wt. %) p-TsOH with 1.5% FA addition at 80 °C, 30 min. The obtained lignin was characterized by FTIR spectroscopy, TGA, 2D HSQC NMR spectroscopy, and GPC. The results indicated that the extracted lignin exhibited excellent properties, such as light color, a low molecular weight (Mw, 5371 g/mol), and a narrow polydispersity (Mw/Mn, 1.63). The pretreated substrate was converted to ethanol via a quasi-simultaneous saccharification and fermentation process (Q-SSF). After fermentation of 60 h, the ethanol concentration reached 38.7 ± 3.3 g/L which was equivalent to a theoretical ethanol yield of 82.9 ± 2.2% based on the glucan content, while the residual glucose concentration was only 4.69 ± 1.4 g/L. In short, this pretreatment strategy protected lignin to form new C–C linkages and improved the enzymatic saccharification of glucan for high-titer ethanol production.Graphical

Cell-wall damage activates DOF transcription factors to promote wound healing and tissue regeneration in Arabidopsis thaliana

Wound healing is a fundamental property of plants and animals that requires recognition of cellular damage to initiate regeneration. In plants, wounding activates a defense response via the production of jasmonic acid and a regeneration response via the hormone auxin and several ethylene response factor (ERF) and NAC domain-containing protein (ANAC) transcription factors. To better understand how plants recognize damage and initiate healing, we searched for factors upregulated during the horticulturally relevant process of plant grafting and found four related DNA binding with one finger (DOF) transcription factors, HIGH CAMBIAL ACTIVITY2(HCA2),TARGET OF MONOPTEROS6(TMO6),DOF2.1,andDOF6, whose expression rapidly activated at the Arabidopsis graft junction. Grafting or wounding a quadruple hca2, tmo6, dof2.1, dof6 mutant inhibited vascular and cell-wall-related gene expression. Furthermore, the quadruple dof mutant reduced callus formation, tissue attachment, vascular regeneration, and pectin methylesterification in response to wounding. We also found that activation of DOF gene expression after wounding required auxin, but hormone treatment alone was insufficient for their induction. However, modifying cell walls by enzymatic digestion of cellulose or pectin greatly enhanced TMO6 and HCA2 expression, whereas genetic modifications to the pectin or cellulose matrix using the PECTIN METHYLESTERASE INHIBITOR5 overexpression line or korrigan1 mutant altered TMO6 and HCA2 expression. Changes to the cellulose or pectin matrix were also sufficient to activate the wound-associated ERF115 and ANAC096 transcription factors, suggesting that cell-wall damage represents a common mechanism for wound perception and the promotion of tissue regeneration.

Structural Changes of Alkali Lignin under Ozone Treatment and Effect of Ozone-Oxidized Alkali Lignin on Cellulose Digestibility

In this study, the structural changes of alkali lignin induced by ozonation were investigated, and the effect of ozone-treated alkali lignin and its mechanism on Avicel enzymatic hydrolysis was examined. The physicochemical properties of alkali lignin were analyzed by FTIR, 1H-13C HSQC NMR, and GPC. It was revealed that ozone pretreatment increased the content of carboxyl and/or aldehyde groups and the negative zeta potential of alkali lignin, which enhanced the electrostatic repulsion between alkali lignin and cellulase; The S/G ratio was reduced, indicating the hydrophobic interaction was diminished. The Langmuir adsorption isotherm showed that the cellulase binding strength of ozone pretreated alkali lignin (OL-pH3, OL-pH7, and OL-pH12 were 16.67, 13.87, and 44.05 mL/g, respectively) was significantly lower than that of alkali lignin (161.29 mL/g). The 72 h hydrolysis yields of Avicel added with OL-pH3, OL-pH7, and OL-pH12 were 55.4%, 58.6%, and 54.9% respectively, which were 2.6–6.3% higher than that of Avicel added with AL (52.3%). This research aimed to reduce the non-productive adsorption between cellulase and lignin by investigating the structural changes of lignin caused by ozone treatment. For the first time, we discovered that ozone-treated alkali lignin has a further promotion effect on the enzymatic digestion of cellulose, providing a green and feasible pretreatment process for the enzymatic hydrolysis of lignocellulose and aiding in the more efficient utilization of biomass.

New Insight into Ultrasonication Pretreatment Prompting Enzymatic Digestibility of Cellulose: Functioning on the Crystalline Structure Transition and Hydrogen Bonds Shift

New Insight into Ultrasonication Pretreatment Prompting Enzymatic Digestibility of Cellulose: Functioning on the Crystalline Structure Transition and Hydrogen Bonds Shift

New Insight into Ultrasonication Pretreatment Prompting Enzymatic Digestibility of Cellulose: Functioning on the Crystalline Structure Transition and Hydrogen Bonds Shift

New Insight into Ultrasonication Pretreatment Prompting Enzymatic Digestibility of Cellulose: Functioning on the Crystalline Structure Transition and Hydrogen Bonds Shift

Enhanced Saccharification of Purple Alfalfa via Sequential Pretreatment with Acidified Ethylene Glycol and Urea/NaOH

Purple Alfalfa is an inexpensive, abundant, readily available lignocellulosic material. This work was attempted to develop an efficient combination pretreatment by sequential HClO4–ethyl glycol–H2O (1.2:88.8:10, w/w/w) extraction at 130 °C in 0.5 h and urea/NaOH (urea 12 wt%, NaOH 7 wt%) soaking at −20 °C for 0.5 h for the pretreatment of purple alfalfa. The porosity, morphology, and crystallinity of pretreated purple alfalfa were characterized with SEM, FM, XRD, and FTIR. This combination pretreatment had a significant influence on hemicellulose removal and delignification. The above changes could enhance cellulose accessibility to enzymes and improve the enzymatic digestibility of cellulose. High yields of reducing sugars from pretreated purple alfalfa were obtained at 93.4%. In summary, this combination pretreatment has high potential application in the future.