Complete valorization of lignocellulose with one-step acidified monophasic phenoxyethanol fractionation.

Effective fractionation of lignocelluosic biomass and subsequent valorization of all three major components under mild conditions were achieved. Pretreatment with acidified monophasic phenoxyethanol (EPH) efficiently removed 92.6% lignin and 80% xylan from poplar at 110 ℃ in 60 min, yielding high-value EPH-xyloside, EPH-modified lignin (EPHL), and a solid residue nearly purely composed of carbohydrates. After removing the grafted acetyl groups using 1% NaOH at 50 ℃, the highest enzymatic digestibility reached 92.3%. EPHL could be recovered in high yield and purity with an uncondensed structure, while xylose was converted to EPH-xyloside, a potential precursor in biomedical industries. Additionally, the acidified monophasic EPH solvent could effectively fractionate biomass from species other than hardwood, achieving over 70% delignification from recalcitrant pinewood under the same mild conditions, demonstrating the high potential of monophasic EPH pretreatment.

Quantitative correlation analysis between particle liquefaction and saccharification through dynamic changes of slurry rheological behavior and particle characteristics during high-solid enzymatic hydrolysis of sugarcane bagasse.

This study identified the intrinsic relationships among slurry rheology, particle characteristics, and lignocellulosic liquefaction/saccharification based on correlation analysis and principal component analysis during the hydrolysis of sugarcane bagasse pretreated by deep eutectic solvents (DES) and mechanical milling (MM). The DES-MM pretreated lignocellulosic slurry (20% solids) exhibited high apparent viscosity of 1.4 × 104 Pa·s and shear stress of 929.0 Pa under steady state. Glucose production had a negative linear correlation with slurry viscosity (R2, 0.69-0.97), whereas its correlation with yield stress (R2, 0.85-0.98) depended on the particle liquefaction rate. The availability of free water provided a major contribution to improving slurry rheology. However, the size reduction of submillimeter particles and the changes in particle hydrophilicity during liquefaction were not significantly correlated with rheological changes. Various interrelated particle characteristics and rheological changes were integrated into two simple principal variables to predict glucose production with a high R2 of 0.96.

Production of Phenylacetylcarbinol via Biotransformation Using the Co-Culture of Candida tropicalis TISTR 5306 and Saccharomyces cerevisiae TISTR 5606 as the Biocatalyst.

Phenylacetylcarbinol (PAC) is a precursor for the synthesis of several pharmaceuticals, including ephedrine, pseudoephedrine, and norephedrine. PAC is commonly produced through biotransformation using microbial pyruvate decarboxylase (PDC) in the form of frozen-thawed whole cells. However, the lack of microorganisms capable of high PDC activity is the main factor in the production of PAC. In addition, researchers are also looking for ways to utilize agro-industrial residues as an inexpensive carbon source through an integrated biorefinery approach in which sugars can be utilized for bioethanol production and frozen-thawed whole cells for PAC synthesis. In the present study, Candida tropicalis, Saccharomyces cerevisiae, and the co-culture of both strains were compared for their biomass and ethanol concentrations, as well as for their volumetric and specific PDC activities when cultivated in a sugarcane bagasse (SCB) hydrolysate medium (SCBHM). The co-culture that resulted in a higher level of PAC (8.65 ± 0.08 mM) with 26.4 ± 0.9 g L-1 ethanol production was chosen for further experiments. Biomass production was scaled up to 100 L and the kinetic parameters were studied. The biomass harvested from the bioreactor was utilized as frozen-thawed whole cells for the selection of an initial pyruvate (Pyr)-to-benzaldehyde (Bz) concentration ([Pyr]/[Bz]) ratio suitable for the PAC biotransformation in a single-phase emulsion system. The initial [Pyr]/[Bz] at 100/120 mM resulted in higher PAC levels with 10.5 ± 0.2 mM when compared to 200/240 mM (8.60 ± 0.01 mM). A subsequent two-phase emulsion system with Pyr in the aqueous phase, Bz in the organic phase, and frozen-thawed whole cells of the co-culture as the biocatalyst produced a 1.46-fold higher PAC level when compared to a single-phase emulsion system. In addition, the cost analysis strategy indicated preliminary costs of USD 0.82 and 1.01/kg PAC for the single-phase and two-phase emulsion systems, respectively. The results of the present study suggested that the co-culture of C. tropicalis and S. cerevisiae can effectively produce bioethanol and PAC from SCB and would decrease the overall production cost on an industrial scale utilizing the two-phase emulsion system with the proposed multiple-pass strategy.

Fractionation of lignin from rice straw using an acidified biphasic solvent system.

To obtain lignin from lignocellulosic biomass, phenoxyethanol (EPH) was employed to construct a biphasic solvent system. The concentration of EPH in this biphasic solvent system was first studied to determine a pretreatment condition for fractionation of lignin. Then, the fractionation of lignin from rice straw was performed under the conditions of temperature 130 °C, cooking time 60 min and sulfuric acid concentration 0.1 M, in 70 % aqueous EPH solvent system. The results showed that 50.97 %, 49.52 % or 82.02 % of the removed lignin with the purity of 89.04 %, 91.30 % or 84.76 % was regenerated from EPH liquor using dimethyl carbonate (DMC), dimethoxymethane (DMM) or diethyl ether (DE) as precipitant, respectively. Additionally, the weight-average molecular weight (Mw) and dispersity index (D) of the regenerated lignin decreased to 4247-4809 g/mol and 1.26-1.60 compared with that of the original lignin (5654 g/mol and 4.78). Finally, the compositional and structural characteristics of lignin, e.g., molecular weight and molecular structure, were also investigated by DSC, HSQC and elemental analysis.

Valorization of rice straw, sugarcane bagasse and sweet sorghum bagasse for the production of bioethanol and phenylacetylcarbinol.

Open burning of agricultural residues causes numerous complications including particulate matter pollution in the air, soil degradation, global warming and many more. Since they possess bio-conversion potential, agro-industrial residues including sugarcane bagasse (SCB), rice straw (RS), corncob (CC) and sweet sorghum bagasse (SSB) were chosen for the study. Yeast strains, Candida tropicalis, C. shehatae, Saccharomyces cerevisiae, and Kluyveromyces marxianus var. marxianus were compared for their production potential of bioethanol and phenylacetylcarbinol (PAC), an intermediate in the manufacture of crucial pharmaceuticals, namely, ephedrine, and pseudoephedrine. Among the substrates and yeasts evaluated, RS cultivated with C. tropicalis produced significantly (p ≤ 0.05) higher ethanol concentration at 15.3 g L-1 after 24 h cultivation. The product per substrate yield (Yeth/s) was 0.38 g g-1 with the volumetric productivity (Qp) of 0.64 g L-1 h-1 and fermentation efficiency of 73.6% based on a theoretical yield of 0.51 g ethanol/g glucose. C. tropicalis grown in RS medium produced 0.303 U mL-1 pyruvate decarboxylase (PDC), a key enzyme that catalyzes the production of PAC, with a specific activity of 0.400 U mg-1 protein after 24 h cultivation. This present study also compared the whole cells biomass of C. tropicalis with its partially purified PDC preparation for PAC biotransformation. The whole cells C. tropicalis PDC at 1.29 U mL-1 produced an overall concentration of 62.3 mM PAC, which was 68.4% higher when compared to partially purified enzyme preparation. The results suggest that the valorization of lignocellulosic residues into bioethanol and PAC will not only aid in mitigating the environmental challenge posed by their surroundings but also has the potential to improve the bioeconomy.

Biphasic fractionation of lignocellulosic biomass based on the combined action of pretreatment severity and solvent effects on delignification.

A novel method based on pretreatment severity and solvent effects on delignification, was introduced to pretreat and fractionate lignocellulose in a 2-phenoxyethanol (EPH) biphasic solvent system. The combined severity factor (CSF) was used to regulate pretreatment severity, and the relative energy difference (RED) of solvent system to lignin was used to evaluate solvent effects. The combined action of pretreatment severity and solvent effects on delignification was first investigated by the response surface regression analysis on the pretreatment of Amorpha. Accordingly, pretreatment and fractionation of Amorpha, poplar and corn straw were then conducted under the optimized conditions. Results showed that >99 % lignin was removed after pretreatment with CSF 3.7845 in a solvent system with RED 0.9371, and 42.94 %, 39.41 % and 70.90 % lignin from Amorpha, poplar and corn straw were respectively regenerated from organosolv liquor after fractionation. Finally, the regenerated products were characterized by FTIR, TG and GPC analysis.

Pretreatment and enzymatic hydrolysis optimization of lignocellulosic biomass for ethanol, xylitol, and phenylacetylcarbinol co-production using Candida magnoliae.

Cellulosic bioethanol production generally has a higher operating cost due to relatively expensive pretreatment strategies and low efficiency of enzymatic hydrolysis. The production of other high-value chemicals such as xylitol and phenylacetylcarbinol (PAC) is, thus, necessary to offset the cost and promote economic viability. The optimal conditions of diluted sulfuric acid pretreatment under boiling water at 95°C and subsequent enzymatic hydrolysis steps for sugarcane bagasse (SCB), rice straw (RS), and corn cob (CC) were optimized using the response surface methodology via a central composite design to simplify the process on the large-scale production. The optimal pretreatment conditions (diluted sulfuric acid concentration (% w/v), treatment time (min)) for SCB (3.36, 113), RS (3.77, 109), and CC (3.89, 112) and the optimal enzymatic hydrolysis conditions (pretreated solid concentration (% w/v), hydrolysis time (h)) for SCB (12.1, 93), RS (10.9, 61), and CC (12.0, 90) were achieved. CC xylose-rich and CC glucose-rich hydrolysates obtained from the respective optimal condition of pretreatment and enzymatic hydrolysis steps were used for xylitol and ethanol production. The statistically significant highest (p ≤ 0.05) xylitol and ethanol yields were 65% ± 1% and 86% ± 2% using Candida magnoliae TISTR 5664. C. magnoliae could statistically significantly degrade (p ≤ 0.05) the inhibitors previously formed during the pretreatment step, including up to 97% w/w hydroxymethylfurfural, 76% w/w furfural, and completely degraded acetic acid during the xylitol production. This study was the first report using the mixed whole cells harvested from xylitol and ethanol production as a biocatalyst in PAC biotransformation under a two-phase emulsion system (vegetable oil/1 M phosphate (Pi) buffer). PAC concentration could be improved by 2-fold compared to a single-phase emulsion system using only 1 M Pi buffer.

A one-step deconstruction-separation organosolv fractionation of lignocellulosic biomass using acetone/phenoxyethanol/water ternary solvent system.

A novel ternary solvent system for organosolv fractionation of lignocellulosic biomass, named APW process, which is composed of acetone, phenoxyethanol and water with the advantages of monophasic deconstruction and biphasic separation of components was developed. Through fractionation of amorpha as a case study, a monophasic APW solution (acetone/phenoxyethanol/water = 5:11:4, volume ratio) with the best lignin affinity was constructed based on Hansen solubility parameters. According to Taguchi experimental design, the optimal conditions were 130 °C, 70 min, 0.15 M sulfuric acid and 20 LSR. Under optimal conditions, removal of lignin and hemicellulose reached 95.60% and 98.39%, respectively. While 80.48% of cellulose was retained in residue and its digestibility was 80.36%. Then, 83.74% of hemicellulose was recovered from aqueous as sugars, and 35.64% of lignin was recovered by precipitation. Moreover, APW process also have effective fractionation of sugarcane bagasse, corn cob and pine, cellulose and hemicellulose recovery were both over 80%.

Effect of a Nonionic Surfactant on Enzymatic Hydrolysis of Lignocellulose Based on Lignocellulosic Features and Enzyme Adsorption.

Reduction in the adsorption of cellulase onto lignin has been thought to be the common reason for the improvement of enzymatic hydrolysis of lignocellulose (EHLC) by a nonionic surfactant (NIS). Few research studies have focused on the relationship between lignocellulosic features and NIS for improving EHLC. This study investigated the impact of Tween20 on the enzymatic hydrolysis and enzyme adsorption of acid-treated and alkali-treated sugarcane bagasse (SCB), cypress, and Pterocarpus soyauxii (PS) with and without being ground. After addition of Tween20, the adsorption of cellulase onto unground and ground alkali-treated SCB increased, and the unground acid-treated SCB exhibited little change in adsorption cellulase, while other unground and ground, treated samples showed decreased cellulase adsorption. Tween20 could improve the enzymatic hydrolysis of acid-treated SCB, while it had little influence on the enzymatic hydrolysis of other treated materials. After being ground, both cellulase adsorption and enzymatic hydrolysis of treated lignocelluloses increased, and Tween20 could enhance the enzymatic hydrolysis of acid-treated materials while hardly affected the enzymatic hydrolysis of alkali-treated materials. This indicated that the promotion effect of Tween20 on enzymatic hydrolysis of treated lignocellulose could not be mainly ascribed to the hindrance of Tween20 to cellulase adsorption on lignin but was related to the lignocellulosic features such as hemicellulose removal and surface morphology changes.

Structural insights reveal the effective Spirulina platensis cell wall dissociation methods for multi-output recovery.

In this work, Spirulina platensis cells harvested in the exponential and equilibrium phases with intact and broken cell walls were treated through a set of alkaline or acidic conditions including alkalis and acids, with solutions of pH 0.0-14.0. The effective Spirulina platensis cell wall dissociation methods for multi-output recovery were obtained. SEM and FTIR were applied to characterize the alkaline and acid treatment details, and Spirulina platensis cell wall dissociation mechanisms, via attacks by OH- or H+, were then proposed. Overall, this study highlights the synthesized multi-output algal product in an integrated strategy with ultracellular structural insight and is valuable for understanding the specific roles of attack ions.

Low-temperature sodium hydroxide pretreatment for ethanol production from sugarcane bagasse without washing process.

A low-temperature sodium hydroxide (NaOH) pretreatment for sugarcane bagasse (SCB) was obtained via the surface response design in this study. However, a large quantity of water consumption and wastewater generation which have been the common problems for alkaline pretreatment of lignocellulose still exists in this pretreatment. In order to reduce water consumption and wastewater generation, this study attempted to perform enzymatic hydrolysis and fermentation of NaOH-treated SCB without washing process. It showed that after pretreatment and solid-liquid separation, NaOH-treated SCB could be directly hydrolysed by cellulase via pH and solid-liquid adjustment without washing steps, and the maximum enzymatic hydrolysis efficiency could reach to 70.2%. A domesticated Saccharomyces cerevisiae Y2034 which can endure 6-times diluted BL was obtained, and realized 67.5% ethanol yield from the enzymatic hydrolysate of unwashed NaOH-treated SCB. It provided a clue for converting NaOH-treated lignocellulose to ethanol at low water consumption and wastewater generation.

Synthesis and properties of porous CLEAs lipase by the calcium carbonate template method and its application in biodiesel production.

In this work, porous cross-linked enzyme aggregates (p-CLEAs) were synthesized by the in situ co-precipitation method using CaCO3 microparticles as templates. The preparation procedure involved the immobilization of crude lipase as CLEAs via precipitation with ammonium sulfate and entrapping these lipase molecules into the CaCO3 templates, followed by DTT (dithiothreitol)-induced assembly of lipase molecules to form lipase microparticles (lipase molecules were assembled into microparticles internally using disulfide bonds within the lipase molecules as the molecular linkers and stimulated by dithiothreitol); finally, the removal of CaCO3 templates was performed by EDTA to form pores in CLEAs. The scanning electron microscopy analysis of p-CLEAs showed a porous structure. p-CLEAs showed obvious improvement in thermal stability (after incubation at 65 °C, p-CLEAs lipase retained 86% relative activity, while free lipase retained only 33.67%) and pH stability (p-CLEAs relative activity was over 90% while for free lipase, the relative activity ranged from 72% to 89% from pH 6 to 9) than free lipase and could hold relatively high activity retention without activity loss at 4 °C for more than six months. The application of p-CLEAs in producing biodiesel showed a higher degree of conversion. The conversion of fatty acid methyl ester (FAME) was 89.7%; this value was higher by approximately 7.4% compared to that of the conventional CLEAs under the optimized conditions of a methanol-oil molar ratio of 6 : 1, with a p-CLEAs lipase dose of 20% and water content of 3% at 45 °C for 24 h. The FAME conversion remained greater than 70% even after reusing the p-CLEAs lipase for 8 reactions. The results demonstrated that the p-CLEAs lipase is suitable for applications in the preparation of biodiesel.

Deep eutectic solvents from hemicellulose-derived acids for the cellulosic ethanol refining of Akebia' herbal residues.

Here, the potential use of herbal residues of Akebia as feedstock for ethanol production is evaluated. Additionally, five deep eutectic solvents from hemicellulose-derived acids were prepared to overcome biomass recalcitrance. Reaction temperatures had more significant influences on solid loss and chemical composition than the molar ratios of choline chloride (ChCl) to derived acids. Glycolic acid resulted in the maximum levels of lignin, xylan and glucan removal, which were 60.0%, 100% and 71.5%, respectively, at 120°C with a 1:6M ratio of ChCl-glycolic acid. In contrast, ChCl-formic acid resulted in the greatest level of glucan retention, at 97.8%, with a lignin removal rate of 40.7% under the same pretreatment conditions. Moreover, ChCl loading could significantly enhance the selectivity of carboxylic acid for lignin dissolution. A 98.0% level of subsequent enzymatic saccharification and a 100% ethanol yield were achieved after ChCl-formic acid pretreatments of Akebia' herbal residues.

Feasibility of reusing the black liquor for enzymatic hydrolysis and ethanol fermentation.

The black liquor (BL) generated in the alkaline pretreatment process is usually thought as the environmental pollutant. This study found that the pure alkaline lignin hardly inhibited the enzymatic hydrolysis of cellulose (EHC), which led to the investigation on the feasibility of reusing BL as the buffer via pH adjustment for the subsequent enzymatic hydrolysis and fermentation. The pH value of BL was adjusted from 13.23 to 4.80 with acetic acid, and the alkaline lignin was partially precipitated. It deposited on the surface of cellulose and negatively influenced the EHC via blocking the access of cellulase to cellulose and adsorbing cellulase. The supernatant separated from the acidified BL scarcely affected the EHC, but inhibited the ethanol fermentation. The 4-times diluted supernatant and the last-time waste wash water of the alkali-treated sugarcane bagasse didn't inhibit the EHC and ethanol production. This work gives a clue of saving water for alkaline pretreatment.

Co-extraction of soluble and insoluble sugars from energy sorghum based on a hydrothermal hydrolysis process.

A process for co-extraction of soluble and insoluble sugars from energy sorghum (ES) was developed based on hydrothermal hydrolysis (HH). Two series of ES were investigated: one (N) with a high biomass yield displayed a higher recalcitrance to sugar release, whereas the second (T) series was characterized by high sugar extraction. The highest total xylose recoveries of 87.2% and 98.7% were obtained for N-11 and T-106 under hydrolysis conditions of 180°C for 50min and 180°C for 30min, respectively. Moreover, the T series displayed higher enzymatic digestibility (ED) than the N series. The high degree of branching (arabinose/xylose ratio) and acetyl groups in the hemicellulose chains of T-106 would be expected to accelerate sugar release during the HH process. In addition, negative correlations between ED and the lignin content, crystallinity index (CrI) and syringyl/guaiacyl (S/G) lignin ratio were observed. Furthermore, finding ways to overcome the thickness of the cell wall and heterogeneity of its chemical composition distribution would make cellulose more accessible to the enzyme.

Structural Changes of Lignin after Liquid Hot Water Pretreatment and Its Effect on the Enzymatic Hydrolysis.

During liquid hot water (LHW) pretreatment, lignin is mostly retained in the pretreated biomass, and the changes in the chemical and structural characteristics of lignin should probably refer to re-/depolymerization, solubilization, or glass transition. The residual lignin could influence the effective enzymatic hydrolysis of cellulose. The pure lignin was used to evaluate the effect of LHW process on its structural and chemical features. The surface morphology of LHW-treated lignin observed with the scanning electron microscopy (SEM) was more porous and irregular than that of untreated lignin. Compared to the untreated lignin, the surface area, total pore volume, and average pore size of LHW-treated lignin tested with the Brunner-Emmet-Teller (BET) measurement were increased. FTIR analysis showed that the chemical structure of lignin was broken down in the LHW process. Additionally, the impact of untreated and treated lignin on the enzymatic hydrolysis of cellulose was also explored. The LHW-treated lignin had little impact on the cellulase adsorption and enzyme activities and somehow could improve the enzymatic hydrolysis of cellulose.

High conversion of sugarcane bagasse into monosaccharides based on sodium hydroxide pretreatment at low water consumption and wastewater generation.

The generation of a great quantity of black liquor (BL) and waste wash water (WWW) has been key problems of the alkaline pretreatment. This work tried to build a sustainable way to recycle the BL for pretreating sugarcane bagasse (SCB) and the WWW for washing the residual solid (RS) of alkali-treated SCB which would be subsequently hydrolysed and fermented. The enzymatic hydrolysis efficiency of the washed RS decreased with the recycling times of BL and WWW increasing. Tween80 at the loading of 0.25% (V/V) could notably improve the enzymatic hydrolysis and had no negative impact on the downstream fermentation. Compared with the non-recycling and BL recycling ways based on alkaline pretreatment, the BL-WWW recycling way could not only maintain high conversion of carbohydrate into monosaccharides and save alkali amount of 45.5%, but also save more than 80% water and generate less than 15% waste water.

Liquid hot water pretreatment of lignocellulosic biomass for bioethanol production accompanying with high valuable products.

Pretreatment is an essential prerequisite to overcome recalcitrance of biomass and enhance the ethanol conversion efficiency of polysaccharides. Compared with other pretreatment methods, liquid hot water (LHW) pretreatment not only reduces the downstream pressure by making cellulose more accessible to the enzymes but minimizes the formation of degradation products that inhibit the growth of fermentative microorganisms. Herein, this review summarized the improved LHW process for different biomass feedstocks, the decomposition behavior of biomass in the LHW process, the enzymatic hydrolysis of LHW-treated substrates, and production of high value-added products and ethanol. Moreover, a combined process producing ethanol and high value-added products was proposed basing on the works of Guangzhou Institute of Energy Conversion to make LHW pretreatment acceptable in the biorefinery of cellulosic ethanol.

Influence of lignin level on release of hemicellulose-derived sugars in liquid hot water.

Lignin layers surrounding hemicelluloses and cellulose in the plant cell walls protect them from deconstruction. This recalcitrance to sugar release is a major limitation for cost-effective industrial conversion of lignocellulosic biomass to biofuels. Many literatures had reported the contribution of lignin removal to cellulose accessibility to enzyme, but less to the hemicellulose hydrolysis. Herein, beech xylan with lignin addition, partly delignified sugarcane bagasse (SB), energy sorghum hybrids (ESH) were treated in liquid hot water (LHW) to investigate the effect of lignin on hemicellulose decomposition. The addition of lignin can enhance the low degree of polymerization of xylooligomers production resulted from the acid catalyzed cleavage of lignin-derived acidic products. However, a negative correlation was observed initially between the lignin level and the total xylose yield from ESH. Furthermore, samples with lignin addition or high lignin content had a great resistant to harsh reaction environment, about 93.5% total xylose lost but only 52.3% released due to the lack of lignin protection for the sample with 100% lignin removal.