Lignin and ash balances of sulfur dioxide-ethanol-water fractionation of sugarcane straw.

Lignin and ash material balances of SO2-ethanol-water (AVAP®) fractionation of sugarcane (SC) straw were thoroughly studied at various conditions. Most of straw lignin and ash dissolve in the liquor and 40-80% of lignin is precipitated after ethanol removal as a pure (∼99%) and sulfur-lean (<2%) fraction. Most of the acid-soluble ash and its elements (Na, K, Fe, Al) as well as large portion of silica are removed from the fiber phase. Straw lignin behavior exhibited differences compared to wood lignin including high apparent content in fiber, higher degree of sulfonation of dissolved lignin, and dense char-like precipitate formation upon ethanol removal. Variation in fractionation conditions did not have significant effect on lignin properties, while post-sulfonation was capable of changing its form from char-like to colloidal precipitate.

Sulfur balance of sulfur dioxide-ethanol-water fractionation of sugarcane straw.

The sulfur balance of SO2-ethanol-water (AVAP®) fractionation of sugarcane (SC) straw was investigated. Hydrogen sulfite and sulfite anions are nearly absent in the liquors, despite cations present in straw, whose effect is thus limited to neutralization of lignosulfonic acids decreasing the acidity. Higher degree of sulfonation was observed for dissolved straw lignin compared to wood lignin (0.8-0.9 vs. 0.25 S/C9). Sulfur dioxide amounts of 0.9-1.2, 4.1-4.3 and 44-49g per o.d.kg straw were bound to pulp, precipitated lignin and lignosulfonic acid, respectively, while the rest of SO2 was recovered by distillation from the spent fractionation suspensions.

Kinetics of SO2-ethanol-water (AVAP®) fractionation of sugarcane straw.

Kinetics of SO2-ethanol-water (AVAP®) fractionation was determined for sugarcane (SC) straw in terms of pulp composition (non-carbohydrate components, cellulose, hemicelluloses) and properties (kappa number, pulp intrinsic viscosity in CED and cellulose degree of polymerization). Effect of temperature (135-165°C) and time (18-118min) was studied at fixed liquor composition (SO2/ethanol/water=12:22.5:65.5, w/w) and a liquor-to-solid ratio (4Lkg(-1)). Interpretation is given in terms of major fractionation reactions, removal of non-carbohydrate components and xylan, as well as acid hydrolysis of cellulose, and is compared to other lignocellulosic substrates (beech, spruce and wheat straw). Overall, SO2-ethanol-water process efficiently fractionates SC straw by separating cellulose from both non-carbohydrate components and xylan while reducing cellulose DP.

The effect of bark on sulfur dioxide-ethanol-water fractionation and enzymatic hydrolysis of forest biomass.

The focus of this study was to find out the effect of bark on SO2-ethanol-water (SEW) fractionation and enzymatic hydrolysis of forest biomass. Softwood bark was found to be more harmful than hardwood bark in both processes. For softwood, the amount of undigested wood in SEW fractionation increased with the increasing bark content, whereas the hardwood bark did not impair the fractionation of wood. The higher the softwood bark content was the lower were the yields in enzymatic hydrolysis likely due to the unproductive binding of enzymes on lignin and other compounds. Addition of surfactant Tween 20 (2% w/w on substrate) prior to enzyme more than doubled the sugar yield of bark-rich softwood pulp. Hardwood bark impaired enzymatic hydrolysis when its share was over 28%. According to a preliminary study, lignosulfonates from the carry-over liquor seem to improve the sugar yield in the enzymatic hydrolysis by acting as a surfactant.

Enzymatic hydrolysis of hardwood and softwood harvest residue fibers released by sulfur dioxide-ethanol-water fractionation.

The enzymatic hydrolysis of hardwood and softwood harvest residues treated by SO2-ethanol-water (SEW) fractionation was studied. The target was to convert these fibers with high yield into glucose monomers which could be further converted into biofuel by a subsequent fermentation stage. Hardwood biomass residues were efficiently digested at low enzyme dosage (5 FPU/g cellulose) whereas the softwood residues required notably higher enzyme dosage (20 FPU) for sufficient conversion. However, cellulase dosage of softwood could be reduced mannanase supplementation. Especially the high lignin content of softwood biomass pulps impairs the digestibility and thereby, improved delignification could notably enhance the hydrolysis yields. It was shown that inferior delignification of SW biomass is due to persistent polyphenolic acids present in coniferous bark, whereas no evidence of the negative effect of inorganics and acetone extractives was observed. Additionally, SW hydrolyzate was successfully converted into a mixture of butanol, acetone and ethanol through ABE fermentation.

Effects of residual lignin and heteropolysaccharides on the bioconversion of softwood lignocellulose nanofibrils obtained by SO2-ethanol-water fractionation.

The amount of residual lignin and hemicelluloses in softwood fibers was systematically varied by SO2-ethanol-water fractionation for integrated biorefinery with nanomaterial and biofuel production. On the basis of their low energy demand in mechanical processing, the fibers were deconstructed to lignocellulose nanofibrils (LCNF) and used as substrate for bioconversion. The effect of LCNF composition on saccharification via multicomponent enzymes was investigated at different loadings. LCNF digestibility was compared with the enzyme activity measured with a quartz crystal microbalance. LCNF hydrolysis rate gradually decreased with lignin and hemicellulose concentration, both of which limited enzyme accessibility. Enzyme inhibition resulted from non-productive binding of proteins onto lignin. Near complete LCNF hydrolysis was achieved, even at high lignin and hemicellulose content. Sugar yields for LCNF were higher than those for precursor SEW fibers, highlighting the benefits of high surface area in LCNF.

Kinetics of SO(2)-ethanol-water (SEW) fractionation of hardwood and softwood biomass.

SO(2)-ethanol-water (SEW) fractionation of forest residues (tree tops, stumps, branches) was investigated to demonstrate the potential of this method for forest biorefineries. The effect of fractionation time on dissolution of wood components was studied. Total mass balances of fractionation show that lignin and hemicelluloses are rapidly dissolved in the spent fractionation liquor whereas cellulose is fully preserved in the solid residue throughout the fractionation treatment. Within 20min treatment at 150°C (SO(2):EtOH:H2O=12:43.5:44.5, by weight, L:W ratio 6Lkg(-1)), 89% of hardwood lignin and 74% of hemicelluloses are dissolved. The corresponding values for softwood biomass are 64% and 74%, respectively, indicating slower delignification but equal hemicellulose removal. Additionally, sulfur content of the feedstocks, solid fractionation residues and spent liquors were analyzed to determine the degree of lignin sulfonation. The obtained results are compared with the stem wood fractionation results.

Oil palm empty fruit bunch to biofuels and chemicals via SO2-ethanol-water fractionation and ABE fermentation.

A process has been developed for conversion of spent liquor produced by SO2-ethanol-water (SEW) fractionation of oil palm empty fruit bunch (OPEFB) fibers to biofuels by ABE fermentation. The fermentation process utilizes Clostridia bacteria that produce butanol, ethanol and acetone solvents at a total yield of 0.26 g/g sugars. A conditioning scheme is developed, which demonstrates that it is possible to utilize the hemicellulose sugars from this agricultural waste stream by traditional ABE fermentation. Fractionation as well as sugar hydrolysis in the spent liquor is hindered by the high cation content of OPEFB, which can be partly removed by acidic leaching suggesting that a better deashing method is necessary. Furthermore, it is inferred that better and more selective lignin removal is needed during conditioning to improve liquor fermentability.

Wood pulp as an immobilization matrix for the continuous production of isopropanol and butanol.

The study was focused on developing a continuous method to produce an alcohol mixture suitable to be used as a gasoline supplement. The immobilized column reactor with wood pulp fibers was successfully used for the continuous production of butanol and isopropanol using Clostridium beijerinckii DSM 6423. A sugar mixture (glucose, mannose, galactose, arabinose and xylose) representing lignocellulose hydrolysate was used as a substrate for the production of solvents. The effect of dilution rate on solvent production was studied during continuous operation. The maximum total solvent concentration of 11.99 g/l was obtained at a dilution rate of 0.16 h(-1). The maximum solvent productivity (5.58 g/l h) was obtained at a dilution rate of 1.5 h(-1). The maximum solvent yield of 0.45 g/g from sugar mixture was observed at 0.25 h(-1). The system will be further used for the solvent production using wood hydrolysate as a substrate.

Efficient fractionation of spruce by SO(2)-ethanol-water treatment: closed mass balances for carbohydrates and sulfur.

SO(2)-ethanol-water (SEW) lignocellulosic fractionation has the potential to overcome the present techno-economic barriers that hinder the commercial implementation of renewable transportation fuel production. In this study, SEW fractionation of spruce wood chips is examined for its ability to separate the main wood components, hemicelluloses, lignin, and cellulose, and the potential to recover SO(2) and ethanol from the spent fractionation liquid. Therefore, overall sulfur and carbohydrate mass balances are established. 95-97 % of the charged SO(2) remains in the liquid and can be fully recovered by distillation. During fractionation, hemicelluloses and lignin are effectively dissolved, whereas cellulose is preserved in the solid (fibre) phase. Hemicelluloses are hydrolysed, producing up to 50 % monomeric sugars, whereas dehydration and oxidation of carbohydrates are insignificant. The latter is proven by the closed carbohydrate material balances as well as by the near absence of corresponding by-products (furfural, hydroxymethylfurfural (HMF) and aldonic acids). In addition, acid methanolysis/GC and acid hydrolysis/high performance anion exchange chromatography (HPAEC) methods for the carbohydrate determination are compared.

Butanol production from lignocellulosics.

Clostridium spp. produce n-butanol in the acetone/butanol/ethanol process. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, the low product yield, toxicity to production strain, multiple-end products and downstream processing of alcohol mixtures. This review describes the use of lignocellulosic feedstocks, bioprocess and metabolic engineering, downstream processing and catalytic refining of n-butanol.

Continuous bio-catalytic conversion of sugar mixture to acetone-butanol-ethanol by immobilized Clostridium acetobutylicum DSM 792.

Continuous production of acetone, n-butanol, and ethanol (ABE) was carried out using immobilized cells of Clostridium acetobutylicum DSM 792 using glucose and sugar mixture as a substrate. Among various lignocellulosic materials screened as a support matrix, coconut fibers and wood pulp fibers were found to be promising in batch experiments. With a motive of promoting wood-based bio-refinery concept, wood pulp was used as a cell holding material. Glucose and sugar mixture (glucose, mannose, galactose, arabinose, and xylose) comparable to lignocellulose hydrolysate was used as a substrate for continuous production of ABE. We report the best solvent productivity among wild-type strains using column reactor. The maximum total solvent concentration of 14.32 g L(-1) was obtained at a dilution rate of 0.22 h(-1) with glucose as a substrate compared to 12.64 g L(-1) at 0.5 h(-1) dilution rate with sugar mixture. The maximum solvent productivity (13.66 g L(-1) h(-1)) was obtained at a dilution rate of 1.9 h(-1) with glucose as a substrate whereas solvent productivity (12.14 g L(-1) h(-1)) was obtained at a dilution rate of 1.5 h(-1) with sugar mixture. The immobilized column reactor with wood pulp can become an efficient technology to be integrated with existing pulp mills to convert them into wood-based bio-refineries.

Continuous acetone-butanol-ethanol fermentation using SO2-ethanol-water spent liquor from spruce.

SO2-ethanol-water (SEW) spent liquor from spruce chips was successfully used for batch and continuous production of acetone, butanol and ethanol (ABE). Initially, batch experiments were performed using spent liquor to check the suitability for production of ABE. Maximum concentration of total ABE was found to be 8.79 g/l using 4-fold diluted SEW liquor supplemented with 35 g/l of glucose. The effect of dilution rate on solvent production, productivity and yield was studied in column reactor consisting of immobilized Clostridium acetobutylicum DSM 792 on wood pulp. Total solvent concentration of 12 g/l was obtained at a dilution rate of 0.21 h(-1). The maximum solvent productivity (4.86 g/l h) with yield of 0.27 g/g was obtained at dilution rate of 0.64 h(-1). Further, to increase the solvent yield, the unutilized sugars were subjected to batch fermentation.

Continuous production of isopropanol and butanol using Clostridium beijerinckii DSM 6423.

Clostridium beijerinckii DSM 6423 was studied using different continuous production methods to give maximum and stable production of isopropanol and n-butanol. In a single-stage continuous culture, when wood pulp was added as a cell holding material, we could increase the solvent productivity from 0.47 to 5.52 g L⁻¹ h⁻¹ with the yield of 54% from glucose. The overall solvent concentration of 7.51 g L⁻¹ (39.4% isopropanol and 60.6% n-butanol) with the maximum solvent productivity of 0.84 g L⁻¹ h⁻¹ was obtained with two-stage continuous culture. We were able to run the process for more than 48 overall retention times without losing the ability to produce solvents.

Production of lactic acid from hemicellulose extracts by Bacillus coagulans MXL-9.

Bacillus coagulans MXL-9 was found capable of growing on pre-pulping hemicellulose extracts, utilizing all of the principle monosugars found in woody biomass. This organism is a moderate thermophile isolated from compost for its pentose-utilizing capabilities. It was found to have high tolerance for inhibitors such as acetic acid and sodium, which are present in pre-pulping hemicellulose extracts. Fermentation of 20 g/l xylose in the presence of 30 g/l acetic acid required a longer lag phase but overall lactic acid yield was not diminished. Similarly, fermentation of xylose in the presence of 20 g/l sodium increased the lag time but did not affect overall product yield, though 30 g/l sodium proved completely inhibitory. Fermentation of hot water-extracted Siberian larch containing 45 g/l total monosaccharides, mainly galactose and arabinose, produced 33 g/l lactic acid in 60 h and completely consumed all sugars. Small amounts of co-products were formed, including acetic acid, formic acid, and ethanol. Hemicellulose extract formed during autohydrolysis of mixed hardwoods contained mainly xylose and was converted into lactic acid with a 94% yield. Green liquor-extracted hardwood hemicellulose containing 10 g/l acetic acid and 6 g/l sodium was also completely converted into lactic acid at a 72% yield. The Bacillus coagulans MXL-9 strain was found to be well suited to production of lactic acid from lignocellulosic biomass due to its compatibility with conditions favorable to industrial enzymes and its ability to withstand inhibitors while rapidly consuming all pentose and hexose sugars of interest at high product yields.

Inhibition effects on fermentation of hardwood extracted hemicelluloses by acetic acid and sodium.

Extraction of hemicellulose from hardwood chips prior to pulping is a possible method for producing ethanol and acetic acid in an integrated forest bio-refinery, adding value to wood components normally relegated to boiler fuel. Hemicellulose was extracted from hardwood chips using green liquor, a pulping liquor intermediate consisting of aqueous NaOH, Na(2)CO(3), and Na(2)S, at 160 degrees C, held for 110 min in a 20 L rocking digester. The extracted liquor contained 3.7% solids and had a pH of 5.6. The organic content of the extracts was mainly xylo-oligosaccharides and acetic acid. Because it was dilute, the hemicellulose extract was concentrated by evaporation in a thin film evaporator. Concentrates from the evaporator reached levels of up to 10% solids. Inhibitors such as acetic acid and sodium were also concentrated by this method, presenting a challenge for the fermentation organisms. Fermentation experiments were conducted with Escherichia coli K011. The un-concentrated extract supported approximately 70% conversion of the initial sugars in 14 h. An extract evaporated down to 6% solids was also fermentable while a 10% solids extract was not initially fermentable. Strain conditioning was later found to enable fermentation at this level of concentration. Alternative processing schemes or inhibitor removal prior to fermentation are necessary to produce ethanol economically.