Overcome Biomass Recalcitrance Research Articles

Enabling Downstream Processing of Lignin Effluent from Cosolvent Enhanced Lignocellulosic Fractionation (CELF) Process through Controlled Electrocatalytic Oxidation and Reduction

Lignin which is a complex organic polymer is abundant in nature. But due to its recalcitrant nature, it can't be converted to value added products easily. Previous studies have found that 40–60 million tons of lignin wastes are generated from the pulp and paper industry globally [1]. The sustainability of these industries can be ensured by converting these wastes into valuable products through environmentally friendly processes. Currently, there are many prevalent catalytic technologies for converting lignin to valuable fuels and chemicals. But these technologies use large amounts of energy and harsh reaction conditions. As a result, catalyst recovery and decomposition often become difficult under such harsh conditions. Besides, bonds between lignin precursors are cleaved unselectively and the selectivity of aromatic products is reduced. An electrocatalytic approach may allow improved control, and few studies have evaluated the electrocatalytic oxidation and reduction of lignin in organic solvents.Organic solvents like tetrahydrofuran (THF) are of interest for lignin electrocatalysis due to their use in the promising cosolvent enhanced lignocellulosic fractionation (CELF) process. Also, THF is a renewable solvent as it can be produced sustainably from the conversion of lignocellulosic biomass. In CELF process, high purity technical lignin can be produced at milder temperatures (1800C) from a mixture of THF and sulfuric acid. Thus, this process has the capability to overcome biomass recalcitrance by breaking β-O-4 aryl ether interunit linkages. Using electrocatalytic conversion processes, additional β-O-4, interunit linkages can be broken down selectively. As a result, if these two processes are integrated, a high amount of phenolic hydroxyl groups with low-content aryl ether linkages will be produced, making the product suitable for the development of biofuels and other chemicals. This study shows that by using controlled electrocatalytic oxidation and reduction in THF/aqueous acidic electrolytes in the ratio of 2:1, nonpolar β-O-4 linkages can be cleaved [2]. This ratio was chosen to mimic the electrolyte composition environment of the CELF pretreatment. The results from both attenuated total reflection-infrared spectroscopy (ATR-IR) and NMR characterization are consistent and show that β-O-4 interlinkage bonds were broken. Quantitative calculations from NMR show that during controlled oxidative potential holds (constant potential oxidation), the presence of aromatic structural components of the lignin polymer increased by 28.67% and aliphatic structural components decreased by 32.73%. On the other hand, during controlled reductive potential holds (constant potential reduction), the presence of aromatic structural compounds decreased by 33.50% and aliphatic structural compounds decreased by 78.43%. These results indicate that electrooxidation and electroreduction may be used strategically to cleave interunit linkages. Finally, it can be said, if electrochemical degradation of lignin is used as a secondary treatment along with the CELF process, it will substantially increase the possibility of transforming lignin into value-added products.References Collins, M. N., Nechifor, M., Tanasă, F., Zănoagă, M., McLoughlin, A., Stróżyk, M. A., Culebras,M., & Teacă, C. A. (2019). Valorization of lignin in polymer and composite systems for advanced engineering applications–a review. International journal of biological macromolecules, 131, 828-849.Hasan, M., Akbari, A., & Greenlee, L. F. (2023). Combined Electrocatalytic Oxidation and Reduction to Selectively Cleave β-O-4 Linkage of Lignin over Platinum Electrode in Organic Solvent: Secondary Treatment Opportunity for CELF Process. ACS Sustainable Chemistry & Engineering. Figure 1

Acidothermus cellulolyticus E1 endoglucanase expressed in planta undergoes extensive hydroxyproline-O-glycosylation and exhibits enhanced impact on biomass digestibility.

E1 holoenzyme was extensively Hyp-O-glycosylated at the proline rich linker region in plants, which substantially increased the molecular size and improved the enzymatic digestibility of the biomass of transgenic plants. Thermophilic E1 endo-1,4-β-glucanase derived from Acidothermus cellulolyticus has been frequently expressed in planta to reconstruct the plant cell wall to overcome biomass recalcitrance. However, the expressed holoenzyme exhibited a larger molecular size (~ 100kDa) than the theoretical one (57kDa), possibly due to posttranslational modifications in the recombinant enzyme within plant cells. This study investigates the glycosylation of the E1 holoenzyme expressed in tobacco plants and determines its impact on enzyme activity and biomass digestibility. The E1 holoenzyme, E1 catalytic domain (E1cd) and E1 linker (E1Lk) were each expressed in tobacco plants and suspension cells. The accumulation of holoenzyme was 2.0- to 2.3- times higher than that of E1cd. The proline-rich E1Lk region was extensively hydroxyproline-O-glycosylated with arabinogalactan polysaccharides. Compared with E1cd, the holoenzyme displayed a broader optimal temperature range (70 to 85 ºC). When grown in greenhouse, the expression of E1 holoenzyme induced notable phenotypic changes in plants, including delayed flowering and leaf variegation post-flowering. However, the final yield of plant biomass was not significantly affected. Finally, plant biomass engineering with E1 holoenzyme showed 1.7- to 1.8-fold higher saccharification efficiency than the E1cd lines and 2.4- to 2.7-fold higher than the wild-type lines, which was ascribed to the synergetic action of the E1Lk and cellulose binding module in reducing cell wall recalcitrance.

Advancing safe bioenergy production from cadmium-contaminated sweet sorghum through mild synergistic pretreatment

Phytoremediation is an eco-friendly and cost-effective method for treating cadmium (Cd)-contaminated soil. However, safe disposal of harvested hyperaccumulators remains scarce. Herein, a mild synergistic pretreatment combining dilute acid and alkali pretreatment was developed for safe biorefining of the Cd-containing sweet sorghum, and Cd was centrally extracted into the acid pretreatment liquid in the first step. The results showed that the synergistic pretreatment was more effective in removing Cd from the biomass and overcoming biomass recalcitrance. During the synergistic pretreatment, the Cd concentration in biomass dramatically decreased from 4.03 to 0.00626 mg/kg, and a considerable amount of lignin and hemicelluloses in biomass was removed, facilitating enzymatic saccharification and safe bioenergy production. The optimal conditions of the synergistic pretreatment resulted in a total sugar yield of 81.6 wt %, which was 4.7-fold higher than that of the original biomass. This strategy holds promise for reducing the cost of industrial saccharification and is a valuable inspiration for the safe handling and resource utilization of Cd-containing biomass.

Microwave-assisted ternary deep eutectic solvent pretreatment for improved rice straw saccharification under mild pretreatment conditions

The development of a green and cost-effective pretreatment method to overcome biomass recalcitrance is a challenging assignment for the efficient valorization of lignocellulosic compounds. In this study, a ternary deep eutectic solvent (DES) consisting of choline chloride (ChCl), p-toluene sulfonic acid (pTSA) and ethylene glycol (EG) was applied in rice straw (RS) pretreatment under microwave (MW) irradiation for the further conversion of the polysaccharide fraction. The effect of the pretreatment temperature, time and pTSA content in the DES on lignin extraction and polysaccharide hydrolysis was assessed by a central composite design (CCD). Under the selected pretreatment conditions (80 ºC, 22.5 min, ChCl:pTSA:EG 1:1:9), 61.6% delignification was achieved, as well as nearly complete conversion of the initial glucan in RS (93.4% glucose yield) and a good xylan conversion rate (48.3% xylose yield), representing a total polysaccharide exploitation of 78.3%. Energetical requirement was remarkably lower compared to conventional conductive heating. Importantly, the ternary DES showed good recyclability pattern after two reuses. Overall, this study demonstrated the potential of MW-assisted ternary DES pretreatment for improved valorization of biomass polysaccharides under mild pretreatment conditions, representing an efficient and energetical-saving alternative pathway within the biorefinery framework.

Semi-Simultaneous Saccharification and Fermentation Improved by Lignin and Extractives Removal from Sugarcane Bagasse

Lignocellulosic biomass and agro-industrial residues are a source of fermentable sugars; however, pretreatments are needed to overcome biomass recalcitrance. This study evaluated the effect of sugarcane bagasse hydrolysis and fermentation in response to dilute acid pretreatment. In natura bagasse, extractive-free bagasse, partially delignified bagasse, and bagasse with added butylated hydroxytoluene antioxidant were pretreated with diluted acid and investigated in semi-simultaneous saccharification and fermentation (S-SSF). The effect of butylated hydroxytoluene (BHT) resulted in lower yields of inhibitors in the liquid fraction of the acid pretreatment (0.01 g L−1 of furfural, 0.01 g L−1 of 5-hydroxymethylfurfural, and 0.68 g L−1 of acetic acid). Partially delignified material and material with BHT resulted in biomass with low hemicellulose and lignin contents, indicating that BHT influenced lignin removal. Extractives removal showed benefits for the acid pretreatment, decreasing the dioxane-soluble material, and a higher yield of glucose and ethanol via S-SSF for the partially delignified material. Enzymatic saccharification of partially delignified material showed 87% of cellulose conversion (24 h with 15 FPU/g), and after 48 h of S-SSF (25 FPU/g), residual 7.06 g L−1 of glucose and production of 15.17 g L−1 of ethanol were observed. The low content of extractives, lignin, and dioxane soluble material resulted in better cellulose accessibility and ethanol yield. Chemical compounds can help remove lignin from biomass favoring ethanol production by S-SSF.

Microbial Lignocellulolytic Enzymes for the Effective Valorization of Lignocellulosic Biomass: A Review

The urgent demand for alternative energy sources has been sparked by the tremendous burden on fossil fuels and the resulting acute energy crisis and climate change issues. Lignocellulosic biomass is a copious renewable and alternative bioresource for the generation of energy fuels and biochemicals in biorefineries. Different pretreatment strategies have been established to overcome biomass recalcitrance and face technological challenges, such as high energy consumption and operational costs and environmental hazards, among many. Biological pretreatment using microbial enzymes is an environmentally benign and low-cost method that holds promising features in the effective pretreatment of lignocellulosic biomass. Due to their versatility and eco-friendliness, cellulases, hemicellulases, and ligninolytic enzymes have been recognized as “green biocatalysts” with a myriad of industrial applications. The current review provides a detailed description of different types of lignocellulolytic enzymes, their mode of action, and their prospective applications in the valorization of lignocellulosic biomass. Solid state fermentation holds great promise in the microbial production of lignocellulolytic enzymes owing to its energy efficient, environment friendly, and higher product yielding features utilizing the lignocellulosic feedstocks. The recent trends in the application of enzyme immobilization strategies for improved enzymatic catalysis have been discussed. The major bottlenecks in the bioprocessing of lignocellulosic biomass using microbial enzymes and future prospects have also been summarized.

In-situ lignin sulfonation for enhancing enzymatic hydrolysis of poplar using mild organic solvent pretreatment

Biomass pretreatment is an essential strategy to overcome biomass recalcitrance and promote lignocellulosic bioconversion. Here, a reusable organic solvent system (formic acid-methanesulfonic acid) was explored to pretreat poplar under a mild temperature (below 100 °C). The results showed that the co-solvent system could extract basically complete hemicelluloses and part of lignin with original cellulose retained in the pretreated substrates. Meanwhile, sulfonic acid groups were introduced into lignin structure remained in the substrates. The glucose conversion yield of the substrates with a higher concentration of sulfonic acid groups (13.2 mmol/kg) reached 45.9 % by reducing the hydrophobic interaction between lignin and cellulase, showing 89.3 % improvement compared with that of the substrates treated with single formic acid. This progressive study aimed to develop a new strategy to realize sulfonation and promote enzymatic hydrolysis of substrates by using mild organic solvent pretreatment.

Understanding the impact of steam pretreatment severity on cellulose ultrastructure, recalcitrance, and hydrolyzability of Norway spruce

The efficient use of softwood in biorefineries requires harsh pretreatment conditions to overcome biomass recalcitrance. Following harsh pretreatments, the hemicellulose is solubilized. Here, we studied the material characteristics of Norway spruce following steam pretreatment at six different severities, relating chemical and structural information to the enzymatic hydrolyzability. Steam pretreatment conditions were defined by two different temperatures (180 °C and 210 °C), with and without the addition of various acids (CH3COOH, H3PO4, H2SO4, SO2). Structural knowledge of the streams is a cornerstone for developing an efficient saccharification process.This study combines advanced structural characterizations to gain fundamental understanding of the influence of severity of pretreatment on spruce. Structural knowledge is a cornerstone in developing an effective saccharification process by modulating pretreatment conditions and enzymes employed.Overall structural properties were assessed by scanning electron microscopy. The effect of stream pretreatment severity on lignin and lignin-carbohydrate bonds was investigated by two-dimensional heteronuclear single quantum correlation nuclear magnetic resonance. Finally, cellulose ultrastructure was studied by applying small/wide-angle X-ray scattering. The structural characteristics of the six pretreated softwood substrates were related to the enzymatic hydrolyzability.With increasing pretreatment severity, surface defibrillation, and lignin depolymeryzation were observed. Further, lignin-carbohydrate complexes signals were detected. Cellulose analysis revealed the rearrangement of microfibrils leading to the formation of larger microfibril aggregates. This microfibril rearrangement likely contributed to the observed increase in enzymatic hydrolysis yields as better enzyme accessibility resulted.

Co-production of levulinic acid and lignin adsorbent from aspen wood with combination of liquid hot water and green-liquor pretreatments

Efficient lignocellulosic fractionation and conversion into platform chemicals provide a considerable opportunity for the industrial operation of a lignocellulosic biorefinery. However, single-step pretreatments are often insufficient to overcome biomass recalcitrance as the existence of insoluble lignin generally restricts component fractionation and conversion. This study performed a two-step pretreatment, consisting of sequential liquid hot water (LHW) and green-liquor (GL), for bioprocessing of aspen biomass into bioproducts such as glucose, levulinic acid (LA), and lignin adsorbent. The two-step pretreated glucan-rich solid displayed high cellulose accessibility, which is responsible for 94% of the glucose yield. The optimized LHW pretreatment (200 °C, 40 min) solubilized 91% of the hemicellulose, which together with monomeric sugars in the hydrolysate solution was straightly heated at 190 °C for 80 min and yielded 72.49% of LA in the presence of sulfuric acid as a catalyst. Moreover, GL post-treatment (Na2CO3+Na2SO3) solution maximized the delignification at 78% from LHW pretreated substrate. The isolated lignin possessed a remarkable ability for Cd2+ and Pb2+ uptake with maximized adsorption values of 250 mg/g and 228 mg/g, respectively. Finally, energy analyses showed that moderate lignocellulosic conversion could obtain an energy recovery of 85%, providing a sustainable pathway for biorefinery approaches.

Characterization of the Biomass Degrading Enzyme GuxA from Acidothermus cellulolyticus.

Microbial conversion of biomass relies on a complex combination of enzyme systems promoting synergy to overcome biomass recalcitrance. Some thermophilic bacteria have been shown to exhibit particularly high levels of cellulolytic activity, making them of particular interest for biomass conversion. These bacteria use varying combinations of CAZymes that vary in complexity from a single catalytic domain to large multi-modular and multi-functional architectures to deconstruct biomass. Since the discovery of CelA from Caldicellulosiruptor bescii which was identified as one of the most active cellulase so far identified, the search for efficient multi-modular and multi-functional CAZymes has intensified. One of these candidates, GuxA (previously Acel_0615), was recently shown to exhibit synergy with other CAZymes in C. bescii, leading to a dramatic increase in growth on biomass when expressed in this host. GuxA is a multi-modular and multi-functional enzyme from Acidothermus cellulolyticus whose catalytic domains include a xylanase/endoglucanase GH12 and an exoglucanase GH6, representing a unique combination of these two glycoside hydrolase families in a single CAZyme. These attributes make GuxA of particular interest as a potential candidate for thermophilic industrial enzyme preparations. Here, we present a more complete characterization of GuxA to understand the mechanism of its activity and substrate specificity. In addition, we demonstrate that GuxA exhibits high levels of synergism with E1, a companion endoglucanase from A. cellulolyticus. We also present a crystal structure of one of the GuxA domains and dissect the structural features that might contribute to its thermotolerance.

Stepwise allomorphic transformations by alkaline and ethylenediamine treatments on bamboo crystalline cellulose for enhanced enzymatic digestibility

Efficient modification and decrystallization of cellulose Iβ in lignocellulosic biomass are potential ways to overcome biomass recalcitrance and facilitates enzymatic degradation. Herein, we have treated moso bamboo and microcrystalline cellulose separately by either alkali or ethylenediamine (EDA) treatments, as well as using both in succession. The pretreatments successfully transformed the initial cellulose Iβ allomorph into cellulose II, IIII, and cellulose IIIII. Crystal structures and crystallinities were determined by Rietveld X-ray diffraction and CP/MAS 13C NMR. Comparatively, the samples that were sequentially treated by EDA and NaOH had low crystallinity (~50%), larger d-spacings of ~4.4 Å for the hydrophobic (110) planes, and less condensed microfibrillar aggregation. The factors simultaneously contributed to as high enzymatic hydrolysis yield as 89.5% and 91.7% respectively, followed by the yields of mercerized cellulose II and cellulose IIIII allomorph. The stepwise allomorphic transformations may provide new strategies on improving efficiency of saccharification process and biomass utilization.

Ethanol Production from Oil Palm Trunk: A Combined Strategy Using an Effective Pretreatment and Simultaneous Saccharification and Cofermentation.

Background Oil palm trunk (OPT) with highly cellulose content is a valuable bioresource for bioethanol production. To produce ethanol from biomass, pretreatment is an essential step in the conversion of lignocellulosic biomass to fermentable sugars such as glucose and xylose. Several pretreatment methods have been developed to overcome biomass recalcitrance. In this study, the effects of different pretreatment methods such as alkali pretreatment, microwave-alkali, and alkaline peroxide combined with autoclave on the lignocellulosic biomass structure were investigated. Moreover, ethanol production from the treated biomass was performed by simultaneous saccharification and cofermentation (SSCF) under different temperatures, fermentation times, and cell ratios of Saccharomyces cerevisiae NCYC 479 and pentose-utilizing yeast, Pichia stipitis NCYC 1541. Results Pretreatment resulted in a significant lignin removal up to 83.26% and cellulose released up to 80.74% in treated OPT by alkaline peroxide combined with autoclave method. Enzymatic hydrolysis of treated OPT resulted in an increase in fermentable sugar up to 93.22%. Optimization of SSCF by response surface method showed that the coculture could work together to produce maximum ethanol (1.89%) and fermentation efficiency (66.14%) under the optimized condition. Conclusion Pretreatment by alkaline peroxide combined with autoclave method and SSCF process could be expected as a promising system for ethanol production from oil palm trunk and various lignocellulosic biomass.

Phylogenetic Occurrence of the Phenylpropanoid Pathway and Lignin Biosynthesis in Plants.

The phenylpropanoid pathway serves as a rich source of metabolites in plants and provides precursors for lignin biosynthesis. Lignin first appeared in tracheophytes and has been hypothesized to have played pivotal roles in land plant colonization. In this review, we summarize recent progress in defining the lignin biosynthetic pathway in lycophytes, monilophytes, gymnosperms, and angiosperms. In particular, we review the key structural genes involved in p-hydroxyphenyl-, guaiacyl-, and syringyl-lignin biosynthesis across plant taxa and consider and integrate new insights on major transcription factors, such as NACs and MYBs. We also review insight regarding a new transcriptional regulator, 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, canonically identified as a key enzyme in the shikimate pathway. We use several case studies, including EPSP synthase, to illustrate the evolution processes of gene duplication and neo-functionalization in lignin biosynthesis. This review provides new insights into the genetic engineering of the lignin biosynthetic pathway to overcome biomass recalcitrance in bioenergy crops.

Effects of Mechanical Refining on Anaerobic Digestion of Dairy Manure.

Mechanical refining (MR) is a cost-effective pretreatment in biochemical conversion processes that is employed to overcome biomass recalcitrance. This work studied the effects of MR on biogas and methane produced by the anaerobic digestion (AD) of dairy manure. The cumulative gas volume and yield from the AD of manure refined at 6k revolutions increased by 33.7 and 7.7% for methane and by 32.0 and 6.4% for biogas, respectively, compared to the unrefined manure. This enhancement was reached by increasing manure solubilization, reducing particle size, and achieving external fibrillation and internal delamination of fibers in manure. However, the highly refined manure (subjected to 60k revolutions) exhibited methane and biogas yields that were reduced by 9.5 and 1.5%, respectively. This decrease was observed because the pore structure was ruptured, and finely ground manure particles were aggregated together at high revolutions (60k), thereby inhibiting the release of organic matter from the manure. Therefore, this study indicates that the MR for pretreatment of dairy manure could have great potential for significantly enhancing AD of dairy manure. Further studies will include optimization of conditions of mechanical refining (i.e., mechanical intensity, process time), a continuous AD of dairy manure pretreated by the MR, and scale-up with cost evaluation.

Carbocation scavenger assisted acid pretreatment followed by mild alkaline hydrogen peroxide (AHP) treatment for efficient production of fermentable sugars and lignin adsorbents from hardwood biomass

Pretreatment is one of the key steps in lignocellulosic biorefinery. Single stage acid pretreatment is often insufficiently satisfactory to overcome biomass recalcitrance, as the presence of insoluble lignin with repolymerized structure always hinders biomass fractionation and saccharification. In this work, a sequential acid-alkali pretreatment was developed to promote biomass fractionation, saccharification and holistic utilization. Results indicated that the addition of 2-naphthol in acid pretreatment enhanced lignin reactivity towards alkali post-treatment. A synergistic effect was observed between 2-naphthol additive and alkali post-treatment in improving biomass saccharification. As a result, up to 90 % of carbohydrates in raw biomass was released and recovered as fermentable sugars after pretreatment and enzymatic hydrolysis. Besides, enzymatic hydrolysis at cellulose loading of 8 % led to a high glucose concentration of 75.51 g/L in enzymatic hydrolysate. Moreover, lignin recovered after alkaline hydrogen peroxide (AHP) treatment and enzymatic hydrolysis both showed great adsorption performance for lead ions, presenting value-added lignin materials. Finally, the mass balance and energy analysis revealed an optimal fractionation of poplar biomass that could achieve an energy recovery of 86 %.

Development of non-derivatizing hydrate salt pre-treatment solvent for pre-treatment and fractionation of corn cob

Major concern in beneficiating lignocellulose is overcoming biomass recalcitrance through pre-treatment. Molten hydrate salts (MHS) is a green solvent with ability to swell and dissolve cellulose and biomass in a non-derivatizing way. Over the last decade, MHSs have been used for isolated cellulose dissolution, however very few studies have been reported on their effectiveness in pre-treating lignocellulosic biomass. Therefore, effectiveness of their application as solvent for pre-treating and fractionating corn cob is presented in this article. In this study, seven molten hydrate salt pre-treatment solvent systems such as unary, binary and ternary mixtures of ZnCl2.4H2O, LiClO4.3H2O and Urea were investigated for their ability to pre-treat and fractionate biomass. The pre-treatment experiments were carried out in a shaking incubator at 70°C for 60 minutes at a biomass: solvent ratio of 1:10. The surface chemistry of the biomass was checked before and after pre-treatment using Fourier Transform infrared spectroscopy. X-ray diffraction and scanning electron microscopy were employed to check the crystallinity and surface morphology of the biomass. Physicochemical analysis consistently indicated a disruption in the structure of corncob due to removal of lignin and hemicellulose during the pre-treatment process. Additionally, results showed a decrease in crystallinity and a change in surface morphology after the pre-treatment using all the seven solvent systems (MHS solvents). The use of ZnCl2.4H2O/ Urea solvent displayed 100% recovery of cellulose, 42% recovery of hemicellulose and 44% recovery of lignin from the corn-cob when compared to the performance of the other proposed solvent systems in this study.

Multiscale modeling and predictive control of cellulose accessibility in alkaline pretreatment for enhanced glucose yield

Even though overcoming biomass recalcitrance through chemical pretreatment significantly improves the glucose yield during biorefinery process, in-depth understanding of the pretreatment has been impeded by the lack of robust kinetic models that capture the microscopic properties of biomass (e.g., cellulose accessibility). In this work, a multiscale model is developed to describe the dynamic evolution of cellulose accessibility during pretreatment. Then, a multi-reaction kinetic model is adopted from the literature and modified to describe the glucose yield of biomass from enzymatic hydrolysis. Then, model approximation techniques are employed to the pretreatment and hydrolysis models to ease the inherent model complexity. Utilizing the approximate models, a model-based feedback controller is designed to enhance the cellulose accessibility while minimizing the heat during alkaline pretreatment. The implementation of the control framework improved the glucose yield by 46% while consuming 7.2% more heat energy compared to a conventional constant-temperature pretreatment method.

Coupling structural characterization with secretomic analysis reveals the mechanism of disruption of the cross-linked structure of bamboo culms

Carboxylesterases, alcohol oxidases and class-II peroxidases were involved in the cleavage of the cross-linked lignin and LCC structures, overcoming biomass recalcitrance.

Overcoming biomass recalcitrance to enhance platform chemical production from soft wood by organosolvolysis coupled with fast pyrolysis

Softwood is an abundantly available lignocelloluse feedstock which can be converted into chemical and liquid fuels via sugar-based platform molecules. However, cost-effective release of pyrolytic sugars from softwood is considerably hindered by the biomass recalcitrance related to its compositions and structures as well as the catalytic effect of alkali and alkaline earth metals. In order to address these challenges, a novel biorefinery based on H2SO4 assisted organosolvolysis of softwood (e.g. pine wood) in high boiling organic solvents coupled with subsequent fast pyrolysis is proposed. The experimental results demonstrated that H2SO4 assisted organosolvolysis could effectively deconstruct pine wood into pentose, organosolv lignin, cellulose-rich fraction, and simutaneously transfer alkali and alkaline earth metals to solutions, thus improving the yields of platform chemcials (levoglucosan and phenols) in subsequent fast pyrolysis. Moreover, different high boiling solvents showed obviously distinct performance for the deconstruction of pine wood and removal of alkali and alkaline earth metals, thus resulting in different yields of platform chemcials in the subsequent fast pyrolysis. The rank order of these solvents which are beneficial for improving the yield of pyrolytic sugars from pine wood was ethylene glycol > glycerin + ethylene glycol (mass ratio of 1:1) > glycerin > γ-valerolactone. The yield of levoglucosan increased drastically from 3.53 wt% of raw pine wood to 27.19 wt% of cellulose-rich fraction pretreated by ethylene glycol with 1 wt% H2SO4. It was found that the yield of levoglucosan from fast pyrolysis of feedstocks was subjecetd to the mutual effect of normalized total alkali and alkaline earth metals’ valencies and severity of delignification. These findings help to provide a simple and efficient process to selective production of platform chemicals from highly recalcitrant biomass.

Overcoming biomass recalcitrance by synergistic pretreatment of mechanical activation and metal salt for enhancing enzymatic conversion of lignocellulose

BackgroundDue to biomass recalcitrance, including complexity of lignocellulosic matrix, crystallinity of cellulose, and inhibition of lignin, the bioconversion of lignocellulosic biomass is difficult and inefficient. The aim of this study is to investigate an effective and green pretreatment method for overcoming biomass recalcitrance of lignocellulose.ResultsAn effective mechanical activation (MA) + metal salt (MAMS) technology was applied to pretreat sugarcane bagasse (SCB), a typical kind of lignocellulosic biomass, in a stirring ball mill. Chlorides and nitrates of Al and Fe showed better synergistic effect with MA, especially AlCl3, ascribing to the interaction between metal salt and oxygen-containing groups induced by MA. Comparative studies showed that MAMS pretreatment effectively changed the recalcitrant structural characteristics of lignocellulosic matrix and reduced the inhibitory action of lignin on enzymatic conversion of SCB. The increase in hydroxyl and carboxyl groups of lignin induced by MAMS pretreatment led to the increase of its hydrophilicity, which could weaken the binding force between cellulase and lignin and reduce the nonproductive binding of cellulase enzymes to lignin.ConclusionsMAMS pretreatment significantly enhanced the enzymatic digestibility of polysaccharides substrate by overcoming biomass recalcitrance without the removal of lignin from enzymatic hydrolysis system.