Glucose Yield Research Articles

Developing a new ethylene glycol/H2O pretreatment system to achieve efficient enzymatic hydrolysis of sugarcane bagasse cellulose and recover highly active lignin: Countercurrent extraction

Improving pretreatment efficiency is a critical premise in achieving efficient biomass conversion, and obtaining high-performance natural polymers is the guarantee of high-value conversion of biomass. In this study, a new pilot-scale continuous countercurrent pretreatment reaction unit about ethylene glycol-alkali solution was designed for pretreating sugarcane bagasse in order to achieve efficient separation of the three major components of lignocellulose when expanding the scale of pretreatment, reduce lignin deposition on the fiber surface, and obtain highly active lignin and excellent enzymatic hydrolysis efficiency of cellulose. X-ray diffractometer (XRD), X-ray photoelectron spectrometer (XPS), brunauer-emmett-teller (BET) and scanning electron microscope (SEM) methods are used to analyze the structural properties of sugarcane bagasse before and after pretreatment, and high-performance liquid chromatography (HPLC) is used to analyze the monosaccharide components in the enzymatic solution. In addition, the structural properties of the recovered lignin are analyzed by gel permeation chromatography (GPC), 31P NMR and 2D-HSQC-NMR methods. The results indicate that the system can gain a high cellulose recovery of 92.99% along with a lignin removal of 95.33%, and recovered lignin has low lignin carbohydrate complexes, low condensation, and rich in phenolic hydroxyl groups for 1.95 mmol/g. Meanwhile, the countercurrent pretreatment system can effectively reduce the deposition of lignin on the cellulose surface, which is evidently superior to the non-countercurrent pretreatment and facilitates the efficiency of enzymatic saccharification of substrate, achieving a high glucose yield of 99% as well as a total sugar yield of 91.11%. The method efficiently separates biomass in a green manner, and solid residues are easily hydrolyzed, showing potential for industrial-scale production.

Comprehensive Wheat Straw Processing with Deep Eutectic Solvent to Deliver Reducing Sugar

Pretreatment is one of the bottlenecks in the cost and energy-efficient biomass valorization. Deep eutectic solvents are potential candidates for being used to address these challenges. In this work, the deep eutectic solvent composed of choline chloride, and acetic acid was studied for its use in wheat straw fractionation. The pretreated biomass was assessed concerning the lignin and glucan content. Under optimized time and temperature conditions, defined using Doehlert matrix chemometric tool, of 3 h 47 min and 139.6 °C, the processed wheat straw contained as much as 42.5 ± 0.42 wt.% and 38.59 ± 1.26 wt.% of glucan and lignin contents, respectively. The need for biomass washing after the pretreatment with deep eutectic solvents and before the enzymatic hydrolysis step was also evaluated. The obtained enzymatic hydrolysis results, i.e., glucan to glucose yield of 27.13 ± 0.25 vs. 25.73 ± 0.08 for washed or unwashed biomass correspondingly, are equally good substrates. Fractal kinetic analysis of the data showed similar values of k and h for both glucose and xylose reactions between washed and unwashed biomass. This confirmed that biomass washing is an unnecessary step, which in turn opens room for biomass processing intensification.

Synergism of jet milling and deep eutectic solvent pretreatment on grapevine lignin fractionation and enhancing enzymatic hydrolysis

Herein, we investigated the synergistic effects of jet milling (JM) and deep eutectic solvent (DES) pretreatment on the fractionation of grapevine lignin and the consequent enhancement of enzymatic hydrolysis. Grapevine, a substantial byproduct of the wine industry, was subjected to JM pretreatment to produce finely powdered particles (median diameter D50 = 98.90), which were then further treated with acidic ChCl-LA and alkaline K2CO3-EG DESs. The results revealed that the combined JM + ChCl-LA pretreatment significantly increased the cellulose preservation under optimal conditions (110 °C, 4 h, and 20 % water content), achieving removal rates of 74.18 % xylan and 66.05 % lignin, respectively. The pretreatment temperature and inhibitor production were reduced, resulting in a remarkable threefold increase in glucose yield compared to untreated samples. Moreover, the structural analysis of the pretreated lignin indicated an enrichment of phenolic units, leading to enhanced antioxidant and antibacterial activities, particularly in the JM pretreated samples. These findings underscore the promising potential of the synergistic JM and DES pretreatment in facilitating the efficient utilization of grapevine lignocellulosic biomass for sustainable biorefinery technologies.

Effect of storage condition on the viability of sago effluents as carbon source in fermentation medium for bioethanol production

In Sarawak, Malaysia, approximately 237 tons/day of sago effluent is commonly discharged into nearby river due to the sago starch extraction process. Due to the high concentration of polymeric compounds, particularly starch, in sago wastewater, which petrifies easily, this condition severely pollutes the environment in the affected area. This study was conducted to determine the viability of using sago effluent as a carbon source and fermentation medium for bioethanol production which indirectly help to minimize the environmental impact as well as the economics of the sago industry. The sago effluent obtained from the local sago mill was analysed for starch content and pH profile while stored at room and cold (4°C) temperature facility. Enzymatic hydrolysis was conducted to convert the residual starch into glucose as carbon source for bioethanol fermentation. Fresh sago effluent can be stored for up to 5 days in cold temperature where the starch content remains constant. The highest starch concentration in sago effluent was 61.33 g/L, in which 50.57 g/L glucose was obtained through the enzymatic hydrolysis process. Hence 82.5% of the starch to glucose conversion yield is revealed. Then, the sago effluent hydrolysate which acts as a carbon source as well as a fermentation medium able to generate 23.14 g/L of bioethanol, displays a 91% theoretical yield of glucose to ethanol. In conclusion, the utilization of sago wastewater as feasible alternative to cheap and locally available and sustainable source of raw materials to produce bioethanol.

Enhancing the dilute acid hydrolysis process using a machine learning approach: investigation of different biomass feedstocks influences glucose and ethanol yields

Enhancing the dilute acid hydrolysis process using a machine learning approach: investigation of different biomass feedstocks influences glucose and ethanol yields

Efficacy and Functional Mechanisms of a Two-Stage Pretreatment Approach Based on Alkali and Ionic Liquid for Bioconversion of Waste Medium-Density Fiberboard.

A novel pretreatment strategy utilizing a combination of NaOH and 1-Butyl-3-methylimidazolium chloride ([Bmim]Cl) was proposed to enhance the enzymatic hydrolysis of abandoned Medium-density fiberboard (MDF). The synergistic effect of NaOH and [Bmim]Cl pretreatment significantly improved the glucose yield, reaching 445.8 mg/g within 72 h, which was 5.04 times higher than that of the untreated samples. The working mechanism was elucidated according to chemical composition, as well as FTIR, 13C NMR, XRD, and SEM analyses. The combined effects of NaOH and [Bmim]Cl led to lignin degradation, hemicellulose removal, the destruction and erosion of crystalline regions, pores, and an irregular microscopic morphology. In addition, by comparing the enzymatic hydrolysis sugar yield and elemental nitrogen content of untreated MDF samples, eucalyptus, and hot mill fibers (HMF), it was demonstrated that the presence of adhesives and additives in waste MDF significantly influences its hydrolysis process. The sugar yield of untreated MDF samples (88.5 mg/g) was compared with those subjected to hydrothermal pretreatment (183.2 mg/g), Ionic liquid (IL) pretreatment (406.1 mg/g), and microwave-assisted ionic liquid pretreatment (MWI) (281.3 mg/g). A long water bath pretreatment can reduce the effect of adhesives and additives on the enzymatic hydrolysis of waste MDF. The sugar yield produced by the combined pretreatment proposed in this study and the removal ability of adhesives and additives highlight the great potential of our pretreatment technology in the recycling of waste fiberboard.

Sustainable wheat straw pretreatment process by self-produced and cyclical crude lactic acid

The purpose of this study was to investigate an environmentally friendly and recyclable pretreatment approach that would enhance the enzymatic digestibility of wheat straw. Wheat straw was pretreated using self-produced crude lactic acid obtained from enzymatic hydrolysate fermentation by Bacillus coagulans. Experimentally, crude lactic acid at low concentration could achieve a pretreatment effect comparable to that of commercial lactic acid. After pretreatment at 180 °C for 60 min with 2.0 % crude lactic acid, hemicellulose could be effectively separated and high recovery of cellulose was ensured, achieving cellulose recovery rate of 95.5 % and hemicellulose removal rate of 92.7 %. Excellent enzymatic hydrolysis was accomplished with a glucose yield of 99.7 %. Moreover, the crude lactic acid demonstrated acceptable pretreatment and enzymatic hydrolysis performance even after three repeated cycles. This not only effectively utilizes the pretreatment solution, but also offers insights into biomass pretreatment using other fermentable acids.

Niobium-based single-atom catalyst promoted fractionation of lignocellulose in choline chloride-lactic acid deep eutectic solvent

Pretreatment is the key step to convert lignocelluloses to sustainable biofuels, biochemicals or biomaterials. In this study, a green pretreatment method based on choline chloride-lactic acid deep eutectic solvent (ChCl-LA) and niobium-based single-atom catalyst (Nb/CN) was developed for the fractionation of corn straw and further enzymatic hydrolysis of cellulose. With this strategy, significant lignin removal of 96.5 % could be achieved when corn straw was pretreated by ChCl-LA (1:2) DES over Nb/CN under 120 °C for 6 h. Enzymatic hydrolysis of the cellulose-enriched fraction (CEF) presented high glucose yield of 92.7 % and xylose yield of 67.5 %. In-depth investigations verified that the high yields of fractions and monosaccharides was attributed to the preliminary fractionation by DES and the deep fractionation by Nb/CN. Significantly, compared to other reported soluble catalysts, the synthesized single-atom catalyst displayed excellent reusability by simple filtration and enzymatic hydrolysis. The recyclability experiments showed that the combination of ChCl-LA DES and Nb/CN could be repeated at least three times for corn straw fractionation, moreover, the combination displayed remarkable feedstock adaptability.

Enhancing fermentable sugar production from sugarcane bagasse through surfactant-assisted ethylene glycol pretreatment and enzymatic hydrolysis: Reduced temperature and enzyme loading

In this study, an efficient coupling surfactant to ethylene glycol pretreatment (EG) and enzymatic hydrolysis of sugarcane bagasse (SCB) was proposed to enhance fermentable sugar production with lower pretreatment energy consumption and reduced enzyme loading. Under optimized conditions, 5 % Tween 80-assisted EG pretreatment of SCB achieved 80.5 % delignification while retaining cellulose (91.6 %) and hemicellulose (81.6 %) content. This led to an enhanced glucose yield of 81.3 %, compared to 65.1 % without Tween 80. The addition of Tween 80 to pretreatment modified residual lignin through etherification, reduced phenolic hydroxyl groups and enhanced hydrophilicity by 23.7 % and 9.4 %, respectively, compared to the lignin sample without Tween 80. Consequently, this modification alleviated non-productive adsorption between lignin and enzymes, improving substrate hydrolyzability. Moreover, when 4.5 % Triton-X 100 was introduced during the hydrolysis of Tween 80-assisted EG-pretreated substrates, a maximum glucose yield of 91.8 % and xylose yield of 92.6 % were achieved. Energy and enzyme cost analyses revealed a reduction of 35 % in pretreatment energy consumption and a 58.8 % decrease in enzyme costs, thanks to the synergistic action of surfactants in EG pretreatment and enzymatic hydrolysis. Overall, integrating surfactants into pretreatment and enzymatic hydrolysis holds promise for highly efficient conversion of SCB into fermentable sugars in enzyme-mediated lignocellulosic biorefineries.

Autohydrolysis of sugarcane bagasse with water reuse: Impacts on residues’ composition and enzymatic hydrolysis

This work presents a new sequential approach to the sugarcane bagasse autohydrolysis process in a way that the liquor from the previous reaction was reused in the next one, and the makeup water for the next batch was used to wash the solid fraction before being added to the liquor in the next batch. This approach was suggested first as a way of reducing the water usage in the process, and second as a way of concentrating the liquor in any interesting component, working with the possibility of losing efficiency towards the glucose production through a cellulose loss to the liquor or enzymatic efficiency loss from a higher inhibitor concentration. Two sets of five sequential batches were performed: one washing the solids with the makeup water prior to the enzymatic step (Set 2), and the other one without the washing step (Set 1), looking forward to the effects of the water reuse over the glucose production and the liquor composition. Autohydrolysis pretreatment removed most of the hemicellulose (∼94 %), with liquor recycling improving its removal until the third batch and stabilizing after that. Although the washing step showed little impact on the composition of the solids, it was determinant to the success of the enzymatic hydrolysis, since it was possible to maintain the cellulose to glucose yield around 53% throughout the batches. There was a 64 % reduction in the water used in the sequential reactions without interfering in the glucose production, which indicates that the proposed strategy could be successfully used to reduce costs and environmental impacts associated with the pretreatment of lignocellulosic biomass.

Unrevealing the influence of reagent properties on disruption and digestibility of lignocellulosic biomass during alkaline pretreatment

Beyond the conventional consideration of pretreatment severity (PS) responsible for biomass disruption, the influence of reagent properties on biomass (LCB) disruption is often overlooked. To investigate the LCB disruption as a function of reagent properties, reagents with distinct cations (NaOH and KOH) and significantly higher delignification potential were chosen. NaOH solution (3 % w/v) with a measured pH of 13.05 ± 0.01 is considered the reference, against which a KOH solution (pH = 13.05 ± 0.01) was prepared for LCB pretreatment under the same PS. Despite comparable lignin content, varying glucose yield of NaOH (68.76 %) and KOH (46.88 %) pretreated residues indicated the presence of heterogeneously disrupted substrate. Holocellulose extracted from raw poplar (ASC, control) and alkaline pretreated residues (C-NaOH and C-KOH) were analyzed using HPLC, XRD, SEM, TGA/DTG, XPS, and 13CP MAS NMR to investigate the pretreatment-induced structural modification. Results revealed that, despite the same pretreatment severity, better disruption in C-NaOH (higher accessible fibril surface and less-ordered region) leading to higher digestibility than C-KOH, likely due to the smaller ionic radius of Na+, facilitates better penetration into dense LCB matrix. This study elucidates the importance of considering the reagent properties during LCB pretreatment, eventually enhancing consciousness while selecting reagents for efficient LCB utilization.

Enhanced starch accumulation in Chlorella sorokiniana as sugar platform and the expression profiling of key regulatory proteins

Microalgae can accumulate considerable starch as a renewable sugar platform for biofuel production. In this study, Chlorella sorokiniana exhibited a significant increase in starch content from 3.0% to 49.5% DM under sulfur limitation (SL) conditions, and the lower the concentration of sulfur was, the more effective it was. Direct enzymatic hydrolysis of wet biomass resulted in a glucose yield of 97.1%, indicating its potential for fermentable sugar production. However, C. sorokiniana grew slowly, and cell division and biomass accumulation decreased under SL conditions, which could be explained by the decreased photosynthetic efficiency and protein content. The sulfur replenishment (SR) confirmed the rapid response of C. sorokiniana in starch accumulation to sulfur availability. Thus, we further investigated the expression profiling of key regulatory proteins under Control-SL-SR stages. The expression of AGPase A was strongly induced in the SL stage and subsequently downregulated during SR stage, suggesting its role as a key regulatory protein for enhanced starch accumulation. Additionally, downregulation of SBE and GBE expression likely contributed to structural modifications of the starch molecules. These proteins could serve as potential targets for metabolic engineering in further improving the starch production capacity in C. sorokiniana.

Functionalized magnetic nanoparticles for Cellic Ctec2 cellulase immobilization: Allowing reusability of enzyme in the conversion of cellulosic biomass

The method of enzyme immobilization can ameliorate the overall stability and restoration of enzymes, hence facilitating their broader application in several sectors. This investigation utilized cellulase as a hydrolytic enzyme. In order to enhance the stability and performance of the cellulase enzyme, the research employed immobilization technology to secure the Cellic Ctec2 cellulase to the synthesized Cs@Fe3O4 nanocomposites. Fe3O4 nanoparticles (NPs) were coated with chitosan obtained from co-precipitation method that served as enzyme carrier. The NPs (Cs@ Fe3O4) were observed under XRD; VSM (vibrating-sample magnetometer) shows saturation magnetizations (Ms), UV–vis, field emission scanning electron microscopy (FESEM), and Fourier transform infrared spectroscopy (FT-IR). Response surface approach was applied to optimize the conditions for immobilization of cellulase. The optimum immobilization of cellulase reaches to 99.1% of loading efficiency and 69.7% of recovery activity with 2.5% of glutaraldehyde concentration. Furthermore, under ideal circumstances the immobilized enzyme’s thermostability, pH stability, temperature tolerance and reusability, were studied with respect to free cellulase. Higher relative activity of cellulase enzyme was observed at pH 5 with 50 °C temperature than free enzyme. One percent CMC hydrolysis is considered for reusability of free and immobilized enzyme and releases 222 mg glucose/g substrate at 24 h, showing great quiescence in cellulosic biomass conversion. Immobilized cellulase demonstrated high reusability by retaining almost 61.2% up to the 5th cycles and 51.2% of activity-maintained 10th cycle of hydrolysis. Reusability of cellulase enzyme can attain a gradual decrease in relative activity as number of repeats of the cycle increases to 10 during hydrolysis and increases in glucose yield after hydrolysis.

Microwave-enhanced hydrolysis of cellulose by acidic ionic liquids

In the [Bmim]Cl reaction medium, five different acidic ionic liquids were used as catalysts to study the effects of reaction time, reaction temperature, system water content, catalyst dosage, microwave power, and other factors on cellulose hydrolysis under microwave irradiation. The results showed that in the [Bmim]Cl reaction system, using N-methylpyrrolidone methylsulfonic acid salt as a catalyst, controlling the microwave reaction time at 10 min, reaction temperature at 130 °C, catalyst dosage at 1 g/g (cellulose), water addition at 0.756 μL/g ([Bmim]Cl), and microwave power at 480 W, resulted in the best cellulose hydrolysis effect with a glucose yield of 74.49%. Compared to conventional heating, the glucose yield increased by 24% and the hydrolysis time was reduced by 77%. Microwave irradiation significantly enhances the cellulose hydrolysis process in an ionic liquid medium.

Sustainability analysis of biorefineries applying biotechnological routes to convert bagasse from non-centrifugal sugar production for rural economic development in Colombia

AbstractPanela is the second most important rural agro-industry after coffee in Colombia. This agro-industry produces a large amount of bagasse from non-centrifugal sugar (B-NCSP) without valorization. B-NCSP is used in combustion boilers in panela production, causing greenhouse gas emissions and health problems. This research aims to compare the sustainability of two B-NCSP biorefineries considering the current residue use. The sustainability of the biorefinery scenarios was analyzed to promote rural economic development in Colombia. In both biorefinery scenarios, biogas was evaluated as an energy vector to meet the energy demand of panela production. Two biorefinery scenarios were considered: (i) saccharification, bioethanol, and biogas production and (ii) acid pretreatment, saccharification, xylitol, bioethanol, and biogas production. Experimental data of the pretreatment, saccharification, and anaerobic digestion stages were used as input information in simulations. Technical, economic, environmental, and social metrics were used to estimate a sustainability index. The experimental glucose yield in scenario 2 was 0.42 ± 0.03 (31.52 ± 2.36 g/L) g/g B-NCSP, 1.8 times greater than scenario 1. The biogas yield in scenario 2 was 504.23 ± 21.68 (71.16% CH4) L/kg B-NCSP, 3.3 times greater than scenario 1. Scenario 1 was unfeasible at different scales. Xylitol production helps to improve the economic feasibility in scenario 2 since the payback period was 15 years. This scenario can create 36 job positions with a wage 18% higher than the minimum wage in Colombia (highest sustainability index of 72.44%). Finally, biogas produced in scenario 2 supplies 62.76% of the energy demand needed in the panela production process, reducing the environmental impact.

Investigating the potential of delignified rice husk as a carbon-rich resource for extracting glucose and its utilization in biocement production through fungal isolates.

Burning rice straw is now a significant issue faced by different regions in India, as its burning releases harmful gases, mainly carbon dioxide. Various techniques are now in trend to utilize the rice straw, e.g., producing compressed natural gas using rice straw, bioethanol, etc., as a substrate for various microorganisms. A high quantity of non-utilized rice husk generates more ideas for its proper utilization. The cellulose, hemicellulose, and lignin found in rice straws can be a fungi growth medium. In this research, the delignification of rice husk is done by acid (2% and 4% H2SO4) and alkali (2% and 4% NaOH) at 121°C at 103kPa for 1h to obtain crude carbon source which is further utilized for biomineralization. The glucose is subjected to qualitative and quantitative analysis using Molisch's and Dinitro salicylic tests. The delignification process showed a positive outcome when 2% H2SO4 is utilized maximum yield of 5.9 ug/ml free sugar concentration. Representing the highest glucose yield compared to the experiment's other acid and base substances used. Various techniques such as field emission-scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and Fourier transformed infra-red (FTIR) spectroscopy are employed to examine surface and chemical alterations. The 2% H2SO4 pretreated rice husk is utilized for microbial-induced calcite precipitation using fungal isolates S1 (3), S1 (18), and S4 (1). The calcite and vaterite produced by biomineralization are confirmed using XRD for fungal isolates namely, S1 (3), S1 (18), and S4 (1) having percentage crystallinity of 59%, 46.428%, and 62.69% percentage crystallinity respectively.

A novel Ruminiclostridium thermocellum cellulase system enhances cellulosic saccharification by elimination of cellobiose feedback inhibition

The Ruminiclostridium thermocellum cellulosome system is one of the most efficient cellulase systems in nature. The main hydrolysis product of cellulosic saccharification by R. thermocellum was cellobiose, which served as a strong feedback inhibitor to the cellulosome. In this study, R. thermocellum M3, which directly hydrolyzes cellulosic biomass into monosaccharides, was used to degrade cellobiose to investigate the kinetic characteristics of cellobiose degradation. Moreover, the transcriptomic characteristics of strain M3 under cellobiose and Avicel culture conditions were compared. The results indicated that strain M3 was able to grow and hydrolyze cellobiose under cellobiose concentrations higher than 20 g/L. The best growth performance and glucose production were obtained under 12 g/L cellobiose. Compared with that of Avicel, cellobiose had greater β-glucosidase activity under cellobiose culture, with a maximum glucose yield of 767.97±19.13 mg/g cellobiose. The transcription results indicated that R. thermocellum M3 resisted the feedback inhibition of cellobiose by regulating genes encoding β-glucosidase, cellobiose phosphorylase and ABC transporters. The high glucose yield of strain M3 was closely related to the expression of the bglX gene. In addition, the gene related to the chemotactic pathway of strain M3 also increased its perception and adaptation to cellobiose.

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.

Enhanced enzymatic sugar production from corn stover by combination of water extraction and glycerol-assisted instant catapult steam explosion

Glycerol-assisted instant catapult steam explosion (ICSE) of lignocellulose is an effective pretreatment method for enhancing sugar production compared to glycerol-free ICSE. In this study, glycerol-assisted ICSE of corn stover was studied in order to understand the reaction mechanisms and further optimize the process. Results showed that water extraction of corn stover prior to ICSE reduced pseudo-lignin formation. The combination of water extraction and glycerol-assisted ICSE led to the formation of lignin with a lower molecular weight (Mw) of 2851 g/mol than 3521 g/mole of that from the combination of water extraction and glycerol-free ICSE. 1H-13C NMR analysis revealed that glycerol likely reacted with lignin carboxylic OHs through esterification while etherification of aliphatic OHs was not observed in ICSE. These lignin analyses indicated that glycerol protected lignin from condensation/repolymerization during glycerol-assisted ICSE. Enzymatic hydrolysis results showed that without water extraction increasing glycerol usage from 0.2 kg/kg stover to 0.4 kg/kg stover improved glucan digestibility to 78% but further increase to 0.5 kg/kg stover reduced glucan digestibility. In addition, at the glycerol usage of 0.2–0.4 kg/kg stover, washing of pretreated stover for removal of glycerol and other biomass-derived compounds did not improve glucan digestibility compared to unwashed ones. Combination of water extraction and glycerol-assisted ICSE led to a high glucan digestibility of 89.7% and a total glucose yield of 25.5 g glucose/100 g stover, which were 30.1% and 7.5 g/100 g stover higher than those derived from glycerol-free ICSE of stover, respectively. Since glycerol is a low-cost carbon source, the resulting enzymatic hydrolysate that contained both glucose and glycerol may be directly used to produce bioproducts by microbial fermentation.Graphical

Unraveling interplay between lignocellulosic structures caused by chemical pretreatments in enhancing enzymatic hydrolysis

To investigate the interplay between substrate structure and enzymatic hydrolysis (EH) efficiency, poplar was pretreated with acidic sodium-chlorite (ASC), 3 % sodium-hydroxide (3-SH), and 3 % sulfuric acid (3-SA), resulting in different glucose yields of 94.10 %, 74.35 %, and 24.51 %, respectively, of pretreated residues. Residues were fractionated into cellulose, lignin and unhydrolyzed residue after EH (for lignin-carbohydrate complex (LCC) analysis) and analyzed using HPLC, FTIR, XPS, CP MAS 13C NMR and 2D-NMR (Lignin and LCC analysis). After delignification, holocellulose exhibited a dramatic increase in glucose yield (74.35 % to 90.82 % for 3-SH and 24.51 % to 80.0 % for 3-SA). Structural analysis of holocellulose suggested the synergistic interplay among cellulose allomorphs to limit glucose yield. Residual lignin analysis from un/pretreated residues indicated that higher β-β' contents and S/G ratios were favorable to the inhibitory effect but unfavourable to the holocellulose digestibility and followed the trend in the following order: 3-SA (L3) > 3-SH (L2) > native-lignin (L1). Analysis of enzymatically unhydrolyzed pretreated residues revealed the presence of benzyl ether (BE1,2) LCC and phenyl glycoside (PG) bond linking to xylose (X) and mannose (M), which yielded a xylan-lignin-glucomannan network. The stability, steric hindrance and hydrophobicity of this network may play a central role in defining poplar recalcitrance.