Plasma-assisted pretreatment of wheat straw.

O3 generated in a plasma at atmospheric pressure and room temperature, fed with dried air (or oxygen-enriched dried air), has been used for the degradation of lignin in wheat straw to optimize the enzymatic hydrolysis and to get more fermentable sugars. A fixed bed reactor was used combined with a CO2 detector and an online technique for O3 measurement in the fed and exhaust gas allowing continuous measurement of the consumption of O3. This rendered it possible for us to determine the progress of the pretreatment in real time (online analysis). The process time can be adjusted to produce wheat straw with desired lignin content because of the online analysis. The O3 consumption of wheat straw and its polymeric components, i.e., cellulose, hemicellulose, and lignin, as well as a mixture of these, dry as well as with 50% water, were studied. Furthermore, the process parameters dry matter content and milled particle size (the extent to which the wheat straw was milled) were investigated and optimized. The developed methodology offered the advantage of a simple and relatively fast (0.5-2 h) pretreatment allowing a dry matter concentration of 45-60%. FTIR measurements did not suggest any structural effects on cellulose and hemicellulose by the O3 treatment. The cost and the energy consumption for lignin degradation of 100 g of wheat straw were calculated.

Upgrading of straw hydrolysate for production of hydrogen and phenols in a microbial electrolysis cell (MEC).

In a microbial electrolysis cell (MEC), hydrolysate produced by hydrothermal treatment of wheat straw was used for hydrogen production during selective recovery of phenols. The average H2 production rate was 0.61 m³ H2/m³ MEC·day and equivalent to a rate of 0.40 kg COD/m³ MEC·day. The microbial community in the anode biofilm was adapted by establishment of xylose-degrading bacteria of the Bacteriodetes phylum (16%) and Geobacter sulfurreducens (49%). During the process, 61% of the chemical oxygen demand was removed as hydrogen at 64% yield. The total energy production yield was 78% considering the energy content in the consumed compounds and the cell voltage of 0.7 V. The highest hydrogen production was equivalent to 0.8 kg COD/m³ MEC·day and was obtained at pH 7-8 and 25°C. Accumulation of 53% w/v phenolic compounds in the liquor was obtained by stepwise addition of the hydrolysate during simultaneous production of hydrogen from consumption of 95% for the hemicellulose and 100% of the fatty acids. Final calculations showed that hydrolysate produced from 1 kg wheat straw was upgraded by means of the MEC to 22 g hydrogen (266 L), 8 g xylan, and 9 g polyphenolics for potential utilization in biobased materials.

Investigation of acetic acid-catalyzed hydrothermal pretreatment on corn stover.

Acetic acid (AA)-catalyzed liquid hot water (LHW) pretreatments on raw corn stover (RCS) were carried out at 195 degrees C at 15 min with the acetic acid concentrations between 0 and 400 g/kg RCS. After pretreatment, the liquor fractions and water-insoluble solids (WIS) were collected separately and tested in terms of the recoveries of glucan and xylan from both the liquor fractions and the WIS, toxicity level of the liquors, and the convertibility of WIS to ethanol. The highest glucan recoveries was found to be 97.42% and 97.94% when 15 and 30 g AA/kg RCS were employed, respectively. The highest xylan recovery of 81.82% was observed by the pretreatment with 10 g AA/kg RCS. The toxic test on liquors showed that the inhibition effect happened to Baker's yeast when the acetic acid used in the pretreatment was higher than 100 g/kg RCS. The WIS obtained from the pretreatment with 15 g and 30 g/kg RCS were subjected to enzymatic hydrolysis and more easily converted to ethanol by Baker's yeast, which gave the ethanol concentration of 33.72 g/L and 32.06 g/L, respectively, higher than 22.04 g/L which was from the non-catalyzed LHW pretreatment (195 degrees C, 15 min). The ethanol concentration from the RCS was only 8.02 g/L.

Feasibility of hydrothermal pretreatment on maize silage for bioethanol production.

The potential of maize silage as a feedstock to produce bioethanol was evaluated in the present study. The hydrothermal pretreatment with five different pretreatment severity factors (PSF) was employed to pretreat the maize silage and compared in terms of sugar recovery, toxic test, and ethanol production by prehydrolysis and simultaneous saccharification and fermentation. After pretreatment, most of the cellulose remained in the residue, ranging between 85.87% by the highest PSF (185 degrees C, 15 min) and 92.90% obtained at the lowest PSF (185 degrees C, 3 min). A larger part of starch, varying from 71.64% by the highest PSF to 78.28% by the lowest, was liberated into liquor part, leaving 8.05-11.74% in the residues. Xylan recovery in the residues increased from 44.25% at the highest PSF to 82.95% at the lowest. The recovery of xylan in liquor changed from 20.13% to 50.33%. Toxic test indicated that all the liquors from the five conditions were not toxic to the Baker's yeast. Pretreatment under 195 degrees C for 7 min had the similar PSF with that of 185 degrees C for 15 min, and both gave the higher ethanol concentration of 19.92 and 19.98 g/L, respectively. The ethanol concentration from untreated maize silage was only 7.67 g/L.

Pretreatment on corn stover with low concentration of formic acid.

Bioethanol derived from lignocellulosic biomass has the potential to replace gasoline. Cellulose is naturally recalcitrant to enzymatic attack, and it also surrounded by the matrix of xylan and lignin, which enhances the recalcitrance. Therefore, lignocellulosic materials must be pretreated to make the cellulose easily degraded into sugars and further fermented to ethanol. In this work, hydrothermal pretreatment on corn stover at 195 degrees for 15 min with and without lower concentration of formic acid was compared in terms of sugar recoveries and ethanol fermentation. For pretreatment with formic acid, the overall glucan recovery was 89% and pretreatment without formic acid yielded the recovery of 94%. Compared with glucan, xylan was more sensitive to the pretreatment condition. The lowest xylan recovery of 55% was obtained after pretreatment with formic acid and the highest of 75% found following pretreatment without formic acid. Toxicity tests of liquor parts showed that there were no inhibitions found for both pretreatment conditions. After simultaneous saccharification and fermentation (SSF) of the pretreated corn stover with Baker's yeast, the highest ethanol yield of 76.5% of the theoretical was observed from corn stover pretreated at 195 degrees for 15 min with formic acid.

Enzymatic hydrolysis and fermentability of corn stover pretreated by lactic acid and/or acetic acid.

Four different pretreatments with and without addition of low concentration organic acids were carried out on corn stover at 195 degrees C for 15 min. The highest xylan recovery of 81.08% was obtained after pretreatment without acid catalyst and the lowest of 58.78% after pretreatment with both acetic and lactic acid. Glucan recovery was less sensitive to the pretreatment conditions than xylan recovery. The pretreatment with acetic and lactic acid yielded the highest glucan recovery of 95.66%. The glucan recoveries of the other three pretreatments varied between 83.92% and 94.28%. Fermentability tests were performed on liquors obtained from all pretreatments and there were no inhibition effect found in any of the liquors. Simultaneous saccharification and fermentation (SSF) of water-insoluble solids (WIS) showed that a high ethanol yield of 88.7% of the theoretical based on glucose in the raw material was obtained following pretreatment at 195 degrees C for 15 min with acetic acid employed. The estimated total ethanol production was 241.1 kg/ton raw material by assuming fermentation of both C-6 and C-5, and 0.51 g ethanol/g sugar.

Identification and characterization of fermentation inhibitors formed during hydrothermal treatment and following SSF of wheat straw.

A pilot plant for hydrothermal treatment of wheat straw was compared in reactor systems of two steps (first, 80 degrees C; second, 190-205 degrees C) and of three steps (first, 80 degrees C; second, 170-180 degrees C; third, 195 degrees C). Fermentation (SSF) with Sacharomyces cerevisiae of the pretreated fibers and hydrolysate from the two-step system gave higher ethanol yield (64-75%) than that obtained from the three-step system (61-65%), due to higher enzymatic cellulose convertibility. At the optimal conditions (two steps, 195 degrees C for 6 min), 69% of available C6-sugar could be fermented into ethanol with a high hemicellulose recovery (65%). The concentration of furfural obtained during the pretreatment process increased versus temperature from 50 mg/l at 190 degrees C to 1,200 mg/l at 205 degrees C as a result of xylose degradation. S. cerevisiae detoxified the hydrolysates by degradation of several toxic compounds such as 90-99% furfural and 80-100% phenolic aldehydes, which extended the lag phase to 5 h. Acetic acid concentration increased by 0.2-1 g/l during enzymatic hydrolysis and 0-3.4 g/l during fermentation due to hydrolysis of acetyl groups and minor xylose degradation. Formic acid concentration increased by 0.5-1.5 g/l probably due to degradation of furfural. Phenolic aldehydes were oxidized to the corresponding acids during fermentation reducing the inhibition level.

Pretreatment of reed by wet oxidation and subsequent utilization of the pretreated fibers for ethanol production.

Common reed (Phragmites australis) is often recognized as a promising source of renewable energy. However, it is among the least characterized crops from the bioethanol perspective. Although one third of reed dry matter is cellulose, without pretreatment, it resists enzymatic hydrolysis like lignocelluloses usually do. In the present study, wet oxidation was investigated as the pretreatment method to enhance the enzymatic digestibility of reed cellulose to soluble sugars and thus improve the convertibility of reed to ethanol. The most effective treatment increased the digestibility of reed cellulose by cellulases more than three times compared to the untreated control. During this wet oxidation, 51.7% of the hemicellulose and 58.3% of the lignin were solubilized, whereas 87.1% of the cellulose remained in the solids. After enzymatic hydrolysis of pretreated fibers from the same treatment, the conversion of cellulose to glucose was 82.4%. Simultaneous saccharification and fermentation of pretreated solids resulted in a final ethanol concentration as high as 8.7 g/L, yielding 73% of the theoretical.

Bioethanol, biohydrogen and biogas production from wheat straw in a biorefinery concept.

The production of bioethanol, biohydrogen and biogas from wheat straw was investigated within a biorefinery framework. Initially, wheat straw was hydrothermally liberated to a cellulose rich fiber fraction and a hemicellulose rich liquid fraction (hydrolysate). Enzymatic hydrolysis and subsequent fermentation of cellulose yielded 0.41 g-ethanol/g-glucose, while dark fermentation of hydrolysate produced 178.0 ml-H(2)/g-sugars. The effluents from both bioethanol and biohydrogen processes were further used to produce methane with the yields of 0.324 and 0.381 m(3)/kg volatile solids (VS)(added), respectively. Additionally, evaluation of six different wheat straw-to-biofuel production scenaria showed that either use of wheat straw for biogas production or multi-fuel production were the energetically most efficient processes compared to production of mono-fuel such as bioethanol when fermenting C6 sugars alone. Thus, multiple biofuels production from wheat straw can increase the efficiency for material and energy and can presumably be more economical process for biomass utilization.

The effect of different substrates and humic acid on power generation in microbial fuel cell operation.

Electricity production from acetate, glucose and xylose with humic acid as mediator was investigated in two chambers microbial fuel cells (MFCs). Acetate produced the highest voltage (570 mV with 1000 Omega) and maximum power density (P(maxd)=123 mW/m(2)) due to a simpler metabolism than with glucose and xylose. Glucose and xylose resulted in P(maxd) of 28 mW/m(2) and 32 mW/m(2) at lower voltage of 380 mV and 414 mV, respectively. P(maxd) increased by 84% and 30%, for glucose and xylose respectively, when humic acid (2g/l) was present in the medium. No significant effect was found with acetate since the internal resistance possessed a limiting effect. The increase of P(maxd) due to humic acid presence was attributed to its ability to act as mediator. Even though pH decreased to 5 with glucose and xylose, due to production of acetate and propionate, the voltage remained on the same level of 250-350 mV.

Wet oxidation pretreatment, enzymatic hydrolysis and simultaneous saccharification and fermentation of clover–ryegrass mixtures

The potential of clover (Trifolium repens) and ryegrass (Lolium perenne) mixtures as raw materials for ethanol production was investigated. Wet oxidation, at 175, 185 or 195 degree Celsius during 10 min at two different oxygen pressures and with either addition or no addition of sodium carbonate, was evaluated as pretreatment method for clover–ryegrass mixtures. The enzymatic hydrolysis of cellulose was significantly improved after pretreatment. The highest conversion efficiency, 93,6 percent, was achieved for the sample pretreated at 195 degree Celsius, 10 min, 1,2 MPa and no addition of Na2CO3. For that sample, the overall glucose yield after pretreatment and hydrolysis was 75,5 percent. No inhibition of cellulose enzymatic conversion by the filtrates was observed. The simultaneous saccharification and fermentation of the pretreated material yielded cellulose conversions of 87,5 and 86,6 percent, respectively, with Saccharomyces cerevisiae and the filamentous fungus Mucor indicus, and revealed that no addition of nutrients is needed for the fermentation of clover–ryegrass hydrolysates(AU)

Hydrothermal treatment of wheat straw at pilot plant scale using a three-step reactor system aiming at high hemicellulose recovery, high cellulose digestibility and low lignin hydrolysis.

A pilot plant (IBUS) consisting of three reactors was used for hydrothermal treatment of wheat straw (120-150 kg/h) aiming at co-production of bioethanol (from sugars) and electricity (from lignin). The first reactor step was pre-soaking at 80 degrees C, the second extraction of hemicellulose at 170-180 degrees C and the third improvement of the enzymatic cellulose convertibility at 195 degrees C. Water added to the third reactor passed countercurrent to straw. The highest water addition (600 kg/h) gave the highest hemicellulose recovery (83%). With no water addition xylose degradation occurred resulting in low hemicellulose recovery (33%) but also in high glucose yield in the enzymatic hydrolysis (72 g/100g glucose in straw). Under these conditions most of the lignin was retained in the fibre fraction, which resulted in a lignin rich residue with high combustion energy (up to 31 MJ/kg) after enzymatic hydrolysis of cellulose and hemicellulose.

Ethanol production from maize silage as lignocellulosic biomass in anaerobically digested and wet-oxidized manure.

In this communication, pretreatment of the anaerobically digested (AD) manure and the application of the pretreated AD manure as liquid medium for the simultaneous saccharification and fermentation (SSF) were described. Furthermore, fermentation of pretreated maize silage and wheat straw was investigated using 2l bioreactors. Wet oxidation performed for 20 min at 121 degrees C was found as the most suitable pretreatment conditions for AD manure. High ammonia concentration and significant amount of macro- and micro-nutrients in the AD manure had a positive influence on the ethanol fermentation. No extra nitrogen source was needed in the fermentation broth. It was shown that the AD manure could successfully substitute process water in SSF of pretreated lignocellulosic fibres. Theoretical ethanol yields of 82% were achieved, giving 30.8 kg ethanol per 100 kg dry mass of maize silage.

Anaerobic biotechnological approaches for production of liquid energy carriers from biomass.

In recent years, increasing attention has been paid to the use of renewable biomass for energy production. Anaerobic biotechnological approaches for production of liquid energy carriers (ethanol and a mixture of acetone, butanol and ethanol) from biomass can be employed to decrease environmental pollution and reduce dependency on fossil fuels. There are two major biological processes that can convert biomass to liquid energy carriers via anaerobic biological breakdown of organic matter: ethanol fermentation and mixed acetone, butanol, ethanol (ABE) fermentation. The specific product formation is determined by substrates and microbial communities available as well as the operating conditions applied. In this review, we evaluate the recent biotechnological approaches employed in ethanol and ABE fermentation. Practical applicability of different technologies is discussed taking into account the microbiology and biochemistry of the processes.

Preliminary results on optimization of pilot scale pretreatment of wheat straw used in coproduction of bioethanol and electricity.

The overall objective in this European Union-project is to develop cost and energy effective production systems for coproduction of bioethanol and electricity based on integrated biomass utilization. A pilot plan reactor for hydrothermal pretreatment (including weak acid hydrolysis, wet oxidation, and steam pretreatment) with a capacity of 100 kg/h was constructed and tested for pretreatment of wheat straw for ethanol production. Highest hemicellulose (C5 sugar) recovery and extraction of hemicellulose sugars was obtained at 190 degrees C whereas highest C6 sugar yield was obtained at 200 degrees C. Lowest toxicity of hydrolysates was observed at 190 degrees C; however, addition of H2O2 improved the fermentability and sugar recoveries at the higher temperatures. The estimated total ethanol production was 223 kg/t straw assuming utilisation of both C6 and C5 during fermentation, and 0.5 g ethanol/g sugar.

High solid simultaneous saccharification and fermentation of wet oxidized corn stover to ethanol.

In this study ethanol was produced from corn stover pretreated by alkaline and acidic wet oxidation (WO) (195 degrees C, 15 min, 12 bar oxygen) followed by nonisothermal simultaneous saccharification and fermentation (SSF). In the first step of the SSF, small amounts of cellulases were added at 50 degrees C, the optimal temperature of enzymes, in order to obtain better mixing condition due to some liquefaction. In the second step more cellulases were added in combination with dried baker's yeast (Saccharomyces cerevisiae) at 30 degrees C. The phenols (0.4-0.5 g/L) and carboxylic acids (4.6-5.9 g/L) were present in the hemicellulose rich hydrolyzate at subinhibitory levels, thus no detoxification was needed prior to SSF of the whole slurry. Based on the cellulose available in the WO corn stover 83% of the theoretical ethanol yield was obtained under optimized SSF conditions. This was achieved with a substrate concentration of 12% dry matter (DM) acidic WO corn stover at 30 FPU/g DM (43.5 FPU/g cellulose) enzyme loading. Even with 20 and 15 FPU/g DM (corresponding to 29 and 22 FPU/g cellulose) enzyme loading, ethanol yields of 76 and 73%, respectively, were obtained. After 120 h of SSF the highest ethanol concentration of 52 g/L (6 vol.%) was achieved, which exceeds the technical and economical limit of the industrial-scale alcohol distillation. The SSF results showed that the cellulose in pretreated corn stover can be efficiently fermented to ethanol with up to 15% DM concentration. A further increase of substrate concentration reduced the ethanol yield significant as a result of insufficient mass transfer. It was also shown that the fermentation could be followed with an easy monitoring system based on the weight loss of the produced CO2.

Thermal wet oxidation improves anaerobic biodegradability of raw and digested biowaste.

Anaerobic digestion of solid biowaste generally results in relatively low methane yields of 50-60% of the theoretical maximum. Increased methane recovery from organic waste would lead to reduced handling of digested solids, lower methane emissions to the environment, and higher green energy profits. The objective of this research was to enhance the anaerobic biodegradability and methane yields from different biowastes (food waste, yard waste, and digested biowaste already treated in a full-scale biogas plant (DRANCO, Belgium)) by assessing thermal wet oxidation. The biodegradability of the waste was evaluated by using biochemical methane potential assays and continuous 3-L methane reactors. Wet oxidation temperature and oxygen pressure (T, 185-220 degrees C; O2 pressure, 0-12 bar; t, 15 min) were varied for their effect on total methane yield and digestion kinetics of digested biowaste. Measured methane yields for raw yard waste, wet oxidized yard waste, raw food waste, and wet oxidized food waste were 345, 685, 536, and 571 mL of CH/g of volatile suspended solids, respectively. Higher oxygen pressure during wet oxidation of digested biowaste considerably increased the total methane yield and digestion kinetics and permitted lignin utilization during a subsequent second digestion. The increase of the specific methane yield for the full-scale biogas plant by applying thermal wet oxidation was 35-40%, showing that there is still a considerable amount of methane that can be harvested from anaerobic digested biowaste.

Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood.

The wet oxidation pretreatment (water, oxygen, elevated temperature, and pressure) of softwood (Picea abies) was investigated for enhancing enzymatic hydrolysis. The pretreatment was preliminarily optimized. Six different combinations of reaction time, temperature, and pH were applied, and the compositions of solid and liquid fractions were analyzed. The solid fraction after wet oxidation contained 58-64% cellulose, 2-16% hemicellulose, and 24-30% lignin. The pretreatment series gave information about the roles of lignin and hemicellulose in the enzymatic hydrolysis. The temperature of the pretreatment, the residual hemicellulose content of the substrate, and the type of the commercial cellulase preparation used were the most important factors affecting the enzymatic hydrolysis. The highest sugar yield in a 72-h hydrolysis, 79% of theoretical, was obtained using a pretreatment of 200 degrees C for 10 min at neutral pH.

Pretreatment of corn stover using wet oxidation to enhance enzymatic digestibility.

Corn stover is an abundant, promising raw material for fuel ethanol production. Although it has a high cellulose content, without pretreatment it resists enzymatic hydrolysis, like most lignocellulosic materials. Wet oxidation (water, oxygen, mild alkali or acid, elevated temperature and pressure) was investigated to enhance the enzymatic digestibility of corn stover. Six different combinations of reaction temperature, time, and pH were applied. The best conditions (60 g/L of corn stover, 195 degrees C, 15 min, 12 bar O2, 2 g/L of Na2CO3) increased the enzymatic conversion of corn stover four times, compared to untreated material. Under these conditions 60% of hemicellulose and 30% of lignin were solubilized, whereas 90% of cellulose remained in the solid fraction. After 24-h hydrolysis at 50 degrees C using 25 filter paper units (FPU)/g of drymatter (DM) biomass, the achieved conversion of cellulose to glucose was about 85%. Decreasing the hydrolysis temperature to 40 degrees C increased hydrolysis time from 24 to 72 h. Decreasing the enzyme loading to 5 FPU/g of DM biomass slightly decreased the enzymatic conversion from 83.4 to 71%. Thus, enzyme loading can be reduced without significantly affecting the efficiency of hydrolysis, an important economical aspect.

Characterization of degradation products from alkaline wet oxidation of wheat straw.

Alkaline wet oxidation pre-treatment (water, sodium carbonate, oxygen, high temperature and pressure) of wheat straw was performed as a 2(4-1) fractional factorial design with the process parameters: temperature, reaction time, sodium carbonate and oxygen. Alkaline wet oxidation was an efficient pre-treatment of wheat straw that resulted in solid fractions with high cellulose recovery (96%) and high enzymatic convertibility to glucose (67%). Carbonate and temperature were the most important factors for fractionation of wheat straw by wet oxidation. Optimal conditions were 10 min at 195 degrees C with addition of 12 bar oxygen and 6.5 g l(-1) Na2CO3. At these conditions the hemicellulose fraction from 100 g straw consisted of soluble hemicellulose (16 g), low molecular weight carboxylic acids (11 g), monomeric phenols (0.48 g) and 2-furoic acid (0.01 g). Formic acid and acetic acid constituted the majority of degradation products (8.5 g). The main phenol monomers were 4-hydroxybenzaldehyde, vanillin, syringaldehyde. acetosyringone (4-hydroxy-3,5-dimethoxy-acetophenone), vanillic acid and syringic acid, occurring in 0.04-0.12 g per 100 g straw concentrations. High lignin removal from the solid fraction (62%) did not provide a corresponding increase in the phenol monomer content but was correlated to high carboxylic acid concentrations. The degradation products in the hemicellulose fractions co-varied with the pre-treatment conditions in the principal component analysis according to their chemical structure, e.g. diacids (oxalic and succinic acids), furan aldehydes, phenol aldehydes, phenol ketones and phenol acids. Aromatic aldehyde formation was correlated to severe conditions with high temperatures and low pH. Apart from CO2 and water, carboxylic acids were the main degradation products from hemicellulose and lignin.