Pretreatment Liquor Research Articles

Enzymatic saccharification promotion for bioenergy poplar under green liquor pretreatment by fully sulfonated polystyrene: Effect of molecular weight

Water-soluble lignin and lignin derivatives are cited to promote the enzymatic saccharification of lignocellulose. Herein, a series of fully sulfonated polystyrene sulfonates (FSPSSs) with various molecular weights (MW) were synthesized through free radical polymerization (FRP) and atom transfer radical polymerization (ATRP) to serve as lignin analogues to boost the enzymatic saccharification of bioenergy poplar under green liquor pretreatment. The FRP-made polymers with MW 944.5 × 103 to 123.6 × 103 g/mol increased the enzymatic hydrolysis digestibility (SED) by 13 % to 18.8 %. On contrary, the ATRP-made polymers with lower MW (3.8 × 103–12.2 × 103 g/mol) showed a weak effect with<8 % improvement in SED. This can be explained the adsorption capacity and the conformation of cellulase-FSPSS complexes, which respond to the reducing nonproductive adsorption correlated to their MWs, due to the strong dependence of molecular conformation on the chain length of strong polyelectrolytes.

High-efficiency recovery and regeneration of choline-polyol deep eutectic solvent for biorefinery via bipolar membrane electrodialysis and ultrafiltration

Deep eutectic solvents (DES) have been widely studied for interesting solvent properties in resources utilization and green conversion of energy. Complex composition of DES and lack of recovery techniques restricts their further scale-up application. Exploring efficient recovery and regeneration methodology of DES in biorefinery could be beneficial for low-carbon circular bioeconomy. Recovery and regeneration of choline-polyol DES (choline chloride-ethylene glycol, ChCl-EG) after biomass pretreatment was studied using bipolar membrane electrodialysis (BMED) with ultrafiltration (UF). UF-BMED treatment worked based on the interception of macromolecular degradation products in pretreatment liquor and regional recovery of Ch+, Cl− with EG. Influence of major parameters on DES recovery performance was studied with emphasis. Maximum recovery ratio of DES reached 97.4% and minimum specific energy consumption of DES recovery approached 6.0 kW·h/kg. Cognition gained from this research revealed an efficient technique for DES recycling after biorefinery.

Valorization of Miscanthus x giganteus for sustainable recovery of anthocyanins and enhanced production of sugars

The increased awareness for eco-friendliness and sustainability has shifted the interest of stakeholders from synthetic colors to natural plant-based pigments. In this study, purple stemmed Miscanthus x giganteus was evaluated as a source of anthocyanins. Hydrothermal pretreatment was studied as a green, chemical-free process for recovering maximum anthocyanins in the pretreatment liquor. The highest recovery of 94.3 ± 1.5% w/w of the total anthocyanin concentration was obtained for a temperature and time combination of 170 °C and 10 min. The pretreatment also improved the enzymatic digestibility of the biomass and led to a 2.1-fold increase in the overall recovery of glucose (70.6 ± 0.5% w/w) at the end of 72 h. The sugar monomers obtained after the enzymatic hydrolysis of the pretreated biomass could be used for the production of biofuels or biochemicals in an integrated biorefinery based on purple-stemmed miscanthus. Overall, this study demonstrates that the clean pretreatment method developed could lead to an additional product stream (rich in anthocyanins) along with its effect in reducing the recalcitrance of miscanthus biomass.

Coproduction of xylooligosaccharides and monosaccharides from hardwood by a combination of acetic acid pretreatment, mechanical refining and enzymatic hydrolysis

Sequential biorefinery treatments of acetic acid (HAC) pretreatment, Papir Forsknings Institutet (PFI) milling and enzymatic hydrolysis were demonstrated for coproduction of xylooligosaccharides (XOS) and fermentable monosaccharides. Results indicated that 36.2% XOS (50.8% X2-X3) and 17.0% low DP xylans were achieved using a HAC pretreatment with a combined severity factor of 0.78. The HAC pretreatment resulted in a XOS-rich prehydrolyzate with a low molecular weight of 1.28 kDa. The endo-xylanase hydrolysis was conducted on the pretreatment liquor to elevate XOS yield and the content of higher-value X2-X3. Moreover, fermentable glucose production from the pretreated residue increased by 2.3 folds when introducing an additional step of PFI refining prior to enzymatic digestion. Properties of substrate including cellulose accessibility, crystallite size, crystalline index and water retention value were in close relationships with enzymatic digestibility. The implementation of proposed biorefinery process will give more insights into the efficient construction of a wood-derived sugar platform.

The effect of soluble phenolic compounds from hydrothermally pretreated wheat straw on Trichoderma reesei cellulases and commercial enzyme cocktails

Hydrothermal methods are commonly applied in pretreatment of lignocellulose for conversion to biofuels and chemicals. This pretreatment partially disassembles and solubilizes hemicelluloses and lignin, releasing sugars and phenolic compounds, which are potential inhibitors for the subsequent enzymatic saccharification step. In this work, the inhibitory effect of soluble phenolics in hydrothermal pretreatment liquor on canonical Trichoderma reesei enzymes was investigated, using purified cellobiohydrolases, endoglucanases, a xylanase, and a lytic polysaccharide monooxygenase, as well as commercial enzyme cocktails Cellic Ctec2 and Celluclast. The pretreatment liquor was fractionated to enrich the phenolics, and thoroughly analyzed. The most sensitive to phenolics inhibition were cellobiohydrolases, the major enzymes in crystalline cellulose degradation and the primary component in cellulolytic mixtures. These were inhibited by the oligophenolics and phenol-carbohydrate complexes (lignin-carbohydrate complex type of compounds), the latter of which could be mitigated by other enzymes, i.e., xylanases and endoglucanases. Addition of lytic polysaccharide monooxygenase to the hydrolysis reactions was found to relieve the negative effect of phenolics. Similarly, hydrolytic activity of a commercial enzyme cocktail Cellic Ctec2 was enhanced in the presence of low concentrations of phenolics.

Reusing the acetone-butanol-ethanol separated broth as the lignocellulose pretreatment liquor for fresh corn stalk biorefinery

In this study, fresh corn stalk was used as the feedstock for the production of bio-butanol via acetone-butanol-ethanol (ABE) fermentation, in which process the wastewater from the first stage sugar containing juice fermentation was recycled by ex situ pervaporation and reused as the pulping liquor of the residual corn stalk bagasse (CSB), affording to effective ABE production in the subsequential second stage fermentation process. Results indicated that the juice fermentation wastewater could effectively depolymerize the CSB. Under optimized conditions of the pulping process (180 °C and 10% (w/v) of biomass loading), 32.45 ± 0.85 g/L of total monomer sugars was obtained in CSB enzymatic hydrolysate, which can be directly used as the substrate for the second-stage cellulosic ABE production. Semi-simultaneous saccharification and fermentation (S-SSF) was carried out using the CSB pulps, so that improving the cellulosic ABE production. 10.94 ± 0.65 g/L of ABE concentration with 0.18 ± 0.01 g/L h of productivity were realized. After hybrid the first-stage juice fermentation and the second-stage CSB biotransformation, total 54.07 ± 1.25 g (ABE) can be produced from 1 kg of fresh corn stalk. The alternative process showed great significance in reduction of the wastewater discharge and ensuring high pulping performances.

Anaerobic digestion of rice straw pretreatment liquor without detoxification for continuous biogas production using a 100 L internal circulation reactor

The production of biogas from lignocellulose by anaerobic digestion (AD) has attracted much attention recently. However, the over-acidification phenomenon and low biogas yield often occur during the AD of lignocellulose. In this work, the liquid-state fermentation mode using pretreatment liquor from dilute acid treated rice straw without detoxification for continuous biogas production in a 100 L internal circulation reactor was studied. The highest biogas yield reached 0.66 m3/kg CODconsumption while the methane yield was 0.38 m3/kg CODconsumption. The pH values of outlet were maintained at 6.9–7.4, and even the inlet pH values were lower than 6.0. Furthermore, from the second day to the end of fermentation, the COD removal rates were more than 90%, even though the inlet COD was close to 20000 mg/L. The organic substances from acid pretreated rice straw were efficiently degraded by AD, which was confirmed by substrate metabolism analysis. These results indicated that the whole fermentation process of AD was very stable and showed great potential for industrial application.

A combination of deep eutectic solvent and ethanol pretreatment for synergistic delignification and enhanced enzymatic hydrolysis for biorefinary process

A novel pretreatment system containing deep eutectic solvents and ethanol (DES-E) for synergistic carbohydrate conversion and delignification was reported in this study. The DES-E pretreatment resulted in an enhanced glucose yield compared to individual DES and ethanol pretreatment for the three tested biomass, including Broussonetia papyrifera, corn stover and pine. To further explore the delignification mechanism, the solubilized lignin and residual lignin from Broussonetia papyrifera was recovered and extracted, then thoroughly characterized. The highest total OH content was found in the DES-E solubilized lignin, which could be used as antioxidant. The presence of ethanol in pretreatment liquor could protect the β-O-4 substructure from breakage and reduce lignin condensation, which favors the subsequent enzymatic hydrolysis. Comparable glucose yield and delignification performance was achieved by recycled DES. DES-E pretreatment offers a promising method for lignin isolation and cellulose digestibility improvement simultaneously.

Combined Extraction and Ethanol Organosolv Fractionation of Exhausted Olive Pomace for Bioactive Compounds

AbstractThe olive pomace oil extracting industry generates large amounts of exhausted olive pomace (EOP), a lignocellulosic waste that needs to be managed according to sustainable criteria. The aim of this work is to devise an integrated strategy to valorize EOP by applying two‐step extraction, and to evaluate the effect of an ethanol organosolv pretreatment on the delignification and enzymatic hydrolysis of the extracted EOP. Once the extraction and organosolv pretreatment conditions are selected, solubilized lignin is recovered from the pretreatment liquor using different methods. In addition to those organosolv lignin samples, a lignin‐rich solid is obtained after enzymatic saccharification of the pretreated solid. All the lignin samples are fully characterized aiming at further valorization. The selected two‐step aqueous extraction (85 °C, 90 min, 10% biomass) removes 89% of the extractives content in raw EOP and achieves the full recovery of phenols and mannitol content in that fraction, 4.7 mg gallic acid equivalents per g EOP and 4.5 mg g−1 EOP, respectively. The organosolv pretreatment (50% ethanol catalyzed with 1% H2SO4, 140 °C, 60 min, 15% biomass) results in a delignified solid with 81% of enzymatic digestibility and a high purity organosolv lignin (&gt;71%), rich in guaiacyl units.

Polystyrene sulfonate is effective for enhancing biomass enzymatic saccharification under green liquor pretreatment in bioenergy poplar

BackgroundWater-soluble lignin (particularly lignosulfonate, LS) has been well documented for its significance on enzymatic saccharification of lignocellulose, though the promotion mechanism has not been fully understood. Much attention has been paid to natural lignin or its derivatives. The disadvantage of using natural lignin-based polymers as promoting agents lies in the difficulty in tailor-incorporating functional groups due to their complex 3D structures. To further improve our understanding on the promotion mechanism of water-soluble lignin in the bioconversion of lignocellulose and to pursue better alternatives with different skeleton structures other than natural lignin, herein we reported a synthetic soluble linear aromatic polymer, sodium polystyrene sulfonate (PSS), to mimic LS for enhancing the efficiency of enzymatic saccharification.ResultsThe role of PSS in enzymatic saccharification of pure cellulose and green liquor-pretreated poplar (GL-P) was explored by analyzing substrate enzymatic digestibility (SED) under different addition dosages and various pH media, along with LS for comparison. At the cellulase loading of 13.3 FPU/g-glucan, the glucose yield of GL-P increased from 53% for the control to 81.5% with PSS addition of 0.1 g/g-substrate. It outperformed LS with the addition of 0.2 g/g-substrate by 6.3%. In the pH range from 4.5 to 6, PSS showed a positive effect on lignocellulose saccharification with the optimum pH at 4.8, where the most pronounced SED of GL-P was achieved. The underlying mechanism was unveiled by measuring zeta potential and using Quartz Crystal Microbalance (QCM) and Multi-parametric Surface Plasmon Resonance (MP-SPR). The results confirmed that the complexes of cellulase and PSS were conjugated and the negatively supercharged complexes reduced non-productive binding effectively along with the improved saccharification efficiency. The thickness of PSS required to block the binding sites of cellulase film was less than half of that of LS, and the PSS adlayer on cellulase film is also more hydrated and with a much lower shear modulus than LS adlayer.ConclusionsPSS as LS analogue is effective for enhancing the biomass enzymatic saccharification of GL-pretreated poplar. PSS exhibited a severer inhibition on the enzymatic saccharification of pure cellulose, while a more positive effect on bioconversion of lignocellulose (GL-P) than LS. In addition, a much lower dosage is required by PSS. The dynamic enzymatic hydrolysis indicated PSS could prolong the processive activity of cellulase. The valid data stemmed from QCM and SPR expressed that PSS bound to cellulases and the as-formed complexes reduced the non-productive adsorption of cellulase onto substrate lignin more efficiently than LS due to its flexible skeleton and highly hydrated structure. Therefore, PSS is a promising alternative promoting agent for lignocellulose saccharification. From another perspective, the synthetic lignin mimics with controllable structures enable us to reach an in-depth understanding of the promotion mechanism of soluble lignins on enzymatic saccharification.

Recovery of low molecular weight compounds from alkaline pretreatment liquor via membrane separations

Lignin is an abundant renewable resource that is a promising substrate for upgrading to fuels and chemicals.

Enzymatic Saccharification Promotion for Bioenergy Poplar Under Green Liquor Pretreatment by Fully Sulfonated Polystyrene: Effect of Molecular Weight

Enzymatic Saccharification Promotion for Bioenergy Poplar Under Green Liquor Pretreatment by Fully Sulfonated Polystyrene: Effect of Molecular Weight

Impact of sulfuric acid pretreatment of durian peel on the production of fermentable sugar and ethanol

After harvesting season, large amounts of durian peels were produced and uselessly disposed of by combustion or landfilling leading to environmental pollution and human health hazards. Proper management of these wastes is necessary to reduce not only an environmental problem but also to create value-added products. Herein, we optimized sulfuric acid pretreatment to promote enzymatic saccharification of durian peels and convert fermentable sugars to bioethanol. Three pretreatment parameters were optimized based on Response Surface Methodology (RSM), including acid concentration (0.5%–3.5%), temperature (60–140 ​°C), and time (20–100 ​min). At optimal pretreatment condition using 2.75% H2SO4, at 127.14 ​°C for 74.13 ​min, 0.53 ​g/g-biomass of reducing sugars were produced, which is 1.88 folds higher than the untreated durian peel. The pretreatment liquor and biomass hydrolysate were analyzed by Gas Chromatograph-Mass spectrometer (GC-MS), and fermentation inhibitors, i.e. acetic acid, furfural, and furan methanol, were identified in those fractions. Due to pretreatment at the optimal condition, a higher yield of reducing sugar was observed, and the production of ethanol from the pretreated biomass was 5.70 ​g/L (equivalent to 87.43% of theoretical yields). These findings demonstrated the potential of using durian waste in the biorefinery concept to achieve a concept of the green economy.

In situ utilization of biomass pretreatment liquor as a novel flocculant for anion dyes removal: Performance and mechanism

In this work, it was first found biomass pretreatment liquor (PL) produced from rice straw (RS) pretreatment with FeCl3 and polyethylene glycol 400 co-solvent can be used in situ as a new flocculant to remove anionic dyes from wastewater. The removal performance of nine dyes was investigated using various PL doses at different pH values. The experiment indicated that the PL had different flocculation effects on these dyes (color removal efficiency: 42.58–99.84%). Positive color removal results for the dyes were unachievable with six commercial coagulants. Among the nine dyes treated by PL flocculation, the best removal efficiencies for color, turbidity and suspended matter were obtained for Congo red. In the flocculation process, Fe3+ plays a role in charge neutralization, lignin nanoparticles (LNP) relies on hydroxyl groups to react instantaneously with the amino groups on the dye, and are bridged together by π–π interactions to promote the formation of floc clusters until they completely settle. Utilization of PL as a flocculant helps pave the way to simultaneously treat waste biomass, waste treatment liquor and dye wastewater. This research is of great significance for future water environment remediation and material development.

All-iron ions mediated electron transfer for biomass pretreatment coupling with direct generation of electricity from lignocellulose

A coupled process of biomass pretreatment for increasing cellulose digestibility and direct conversion of biomass to electricity has been developed with ferric or ferricyanide ions as the anode electron carriers, and Fe(NO3)3 activated by HNO3 as the cathode electron carriers. Pretreated substrates are subjected to enzymatic hydrolysis for release of fermentable sugars, while the pretreatment liquor is employed as anolyte for electricity generation in a liquid flow fuel cell (LFFC). Pretreatment of sugarcane bagasse with 2 M FeCl3 in anode reactor removes ∼ 100% hemicelluloses and obtains 76% enzymatic glucan conversion (EGC), while pretreatment with 0.1 M K3[Fe(CN)6] in 0.5 M KOH achieves 78% lignin removal, 95.8% EGC and 85.1% xylan conversion. From 1000 g bagasse, 171.3 g fermentable sugars is produced with co-generation of 101.4 W·h electricity based on FeCl3 pretreatment, while 519 g fermentable sugars and 28.9 W·h electricity are obtained based on K3[Fe(CN)6] pretreatment.

Apple pomace biorefinery for ethanol, mycoprotein, and value-added biochemicals production by Mucor indicus

Apple pomace, a waste resulting after juice extraction, is often discarded in landfills, which can cause environmental issues. However, since apple pomace contains high levels of free sugars and structural carbohydrates, its conversion to fuels and biomaterials can mitigate environmental issues. In this study, an apple pomace feedstock-based biorefinery utilizing ethanolic Organosolv pretreatment and simultaneous saccharification and fermentation (SSF) was conceptualized. Ethanolic based Organosolv pretreatment was performed at 24 different conditions to find the optimum conditions for maximum ethanol production. The resulting pretreated solids and pretreatment liquor were fermented to ethanol and/or biomass with filamentous fungi Mucor indicus. The pretreatment of apple pomace at 100 °C for 30 min with 50% ethanol and 0.5% sulfuric acid resulted in the highest ethanol yield of 244.9 g/kg of apple pomace. This yield included 47.2 g ethanol from pretreated solids and 197.7 g from pretreatment liquor, where the corresponding amount of pretreatment liquor and solids were subjected to fermentation. However, the aerobic fermentation of pretreatment liquor resulted in lower ethanol yield (117.4 g/kg of apple pomace) than the fermentation under anaerobic conditions, but high levels of mycoprotein (29.0 g/kg of apple pomace) and chitin and chitosan (8.7 g/kg of apple pomace) were produced.

Rapid and efficient microwave-assisted guanidine hydrochloride deep eutectic solvent pretreatment for biological conversion of castor stalk

Microwave-assisted guanidine hydrochloride deep eutectic solvents (DESs) was developed for rapid and efficient pretreatment of castor stalk. The DES synthesized with guanidine hydrochloride and lactic acid showed a better delignification (92.02%) and enzymatic saccharification yield (96.3%) than choline chloride and lactic acid DES resulted. In addition, high-purity (up to 98%) lignin was recovered from the pretreatment liquor. The good recyclability of the guanidine hydrochloride-based DES was also proven with up to 90% cellulose hydrolysis with third-time recycled DES without post purification. The proposed microwave-assisted guanidine hydrochloride/lactic acid DES showed its great potentials as a highly effective and recyclable pretreatment solvent for future biorefinery strategies.

Efficient production of xylooligosaccharides rich in xylobiose and xylotriose from poplar by hydrothermal pretreatment coupled with post-enzymatic hydrolysis

A promising approach for production of value-added xylooligosaccharides (XOS) from poplar was developed by combining hydrothermal pretreatment and endo-xylanase post-hydrolysis. Results showed that the 35.4% XOS (DP 2–6) and 17.6% low DP xylans (DP > 6) were obtained at the identified optimal condition (170 °C, 50 min) for hydrothermal pretreatment. Structural features of low DP xylans generated during the hydrothermal pretreatment were examined, revealing that low DP xylans are mainly comprised of 4-O-methylglucuronic xylan and are involved in lignin carbohydrate complexes. Moreover, higher pretreatment intensity promoted the cleavage of side-chain substituents including arabinose and glucuronic acid groups. The subsequent endo-xylanase hydrolysis of the pretreatment liquor hydrolyzed low DP xylans, contributing to a significant improvement in xylobiose and xylotriose proportions. This combined strategy resulted in a XOS with conversion yield of 44.6% containing 78.7% xylobiose and xylotriose starting from the initial xylan in raw poplar.

Hydrothermal pretreatment: An efficient process for improvement of biobutanol, biohydrogen, and biogas production from orange waste via a biorefinery approach

Orange waste (OW), an abundant and severe globally environmental treat, was used for biobutanol and biohydrogen production emploing acetone-butanol-ethanol (ABE) fermentation through a biorefinery process. The solvent yield from untreated OW was insufficient; thus, the substrate was subjected to hydrothermal pretreatment before hydrolysis. The pretreatment at 140 ℃ for 30 min resulted in the solid with the highest yield of hydrolysis and fermentation. Moreover, the anaerobic digestion of hydrolysis residue produced appreciable amounts of biomethane. However, the pretreatment liquor was not fermentable; thus, it was detoxified by overliming for 24 h at 30 ℃ and then fermented. Overall, this sustainable biorefinery, based on pretreatment without any additional chemical agent, hydrolysis of pretreated solids, detoxification of pretreatment liquor, ABE fermentation, and anaerobic digestion of residues, produced 42.3 g biobutanol, 33.1 g acetone, 13.4 g ethanol, 104.5 L biohydrogen, and 28.3 L biomethane per kg of OW that contained 4560 kJ energy.

Rice straw pretreatment with KOH/urea for enhancing sugar yield and ethanol production at low temperature

In this experiment, a low-temperature (< 100 °C) KOH/urea (KU) pretreatment of rice straw (RS) was performed to enhance its bioconversion to ethanol. The pretreatment factors were optimized by Box-Behnken Design of response surface method (RSM), resulting in an optimum KU concentration of 3%, with a solid to liquid ratio of 1:15 for 3 h at 70 °C. Under the above conditions, five-times higher sugar yields (92.38 %) and six-times higher ethanol production (87.13 %) compared with the control experiment were obtained. The cell wall composition and crystallinity index (CrI) of RS changed significantly after KU pretreatment, with cellulose amounts raised, while the CrI increased and the amounts of hemicellulose and lignin decreased. Additionally, the pretreatment liquor is high in potassium and nitrogen contents, which may be used as a fertilizer for plant cultivation to achieve a zero-emission biorefinery process.