Improved bio-energy yields via sequential ethanol fermentation and biogas digestion of steam exploded oat straw

Using standard laboratory equipment, thermochemically pretreated oat straw was enzymatically saccharified and fermented to ethanol, and after removal of ethanol the remaining material was subjected to biogas digestion. A detailed mass balance calculation shows that, for steam explosion pretreatment, this combined ethanol fermentation and biogas digestion converts 85-87% of the higher heating value (HHV) of holocellulose (cellulose and hemicellulose) in the oat straw into biofuel energy. The energy (HHV) yield of the produced ethanol and methane was 9.5-9.8 MJ/(kg dry oat straw), which is 28-34% higher than direct biogas digestion that yielded 7.3-7.4 MJ/(kg dry oat straw). The rate of biogas formation from the fermentation residues was also higher than from the corresponding pretreated but unfermented oat straw, indicating that the biogas digestion could be terminated after only 24 days. This suggests that the ethanol process acts as an additional pretreatment for the biogas process.     

Enhancement of enzymatic saccharification of cellulose by cellulose dissolution pretreatments

Attempts were made to enhance cellulose saccharification by cellulase using cellulose dissolution as a pretreatment step. Four cellulose dissolution agents, NaOH/Urea solution, N-methylmorpholine-N-oxide (NMMO), ionic liquid (1-butyl-3-methylimidazolium chloride; [BMIM]Cl) and 85% phosphoric acid were employed to dissolve cotton cellulose. In comparison with conventional cellulose pretreatment processes, the dissolution pretreatments were operated under a milder condition with temperature <130 °C and ambient pressure. The dissolved cellulose was easily regenerated in water. The regenerated celluloses exhibited a significant improvement (about 2.7- to 4.6-fold enhancement) on saccharification rate during 1st h reaction. After 72 h, the saccharification yield ranged from 87% to 96% for the regenerated celluloses while only around 23% could be achieved for the untreated cellulose. Even with high crystallinity, cellulose regenerated from phosphoric acid dissolution achieved the highest saccharification rates and yield probably due to its highest specific surface area and lowest degree of polymerization (DP).     

Effect of biological pretreatments in enhancing corn straw biogas production

A biological pretreatment with new complex microbial agents was used to pretreat corn straw at ambient temperature (about 20°C) to improve its biodegradability and anaerobic biogas production. A complex microbial agent dose of 0.01% (w/w) and pretreatment time of 15 days were appropriate for biological pretreatment. These treatment conditions resulted in 33.07% more total biogas yield, 75.57% more methane yield, and 34.6% shorter technical digestion time compared with the untreated sample. Analyses of chemical compositions showed 5.81-25.10% reductions in total lignin, cellulose, and hemicellulose contents, and 27.19-80.71% increases in hot-water extractives; these changes contributed to the enhancement of biogas production. Biological pretreatment could be an effective method for improving biodegradability and enhancing the highly efficient biological conversion of corn straw into bioenergy.     

Swelling and dissolution of cellulose. Part IV: Free floating cotton and wood fibres in ionic liquids

The objective of this paper is to investigate if the swelling and dissolution mechanisms found for aqueous solvents are valid for non-aqueous ones. Three different ionic liquids were used and the swelling and dissolution mechanisms were investigated by optical methods. Native and enzymatically treated cellulose fibres (cotton and wood fibres) are dipped into three ionic liquids (1-N-butyl-3-methylimidazolium chloride ([C4mim]+Cl−)/DMSO, allylmethylimidazolium bromide ([Amim]+Br−) and butenylmethylimidazolium bromide ([Bmim]+Br−). ([C4mim]+Cl−)/DMSO shows a swelling of cellulose by ballooning and then dissolution. ([Amim]+Br−) and ([Bmim]+Br−) show a homogeneous swelling but no dissolution. The swelling and dissolution mechanisms of cellulose in ionic liquids are similar to those observed in aqueous solvents. It indicates that the swelling and dissolution mechanisms are entirely due to the way cellulose fibres are structured, not depending on the type of solvent. The quality of the solvent is giving the type of mechanism.     

Enhancing the anaerobic digestion of corn stalks using composite microbial pretreatment

A composite microbial system (XDC-2) was used to pretreat and hydrolyze corn stalk to enhance anaerobic digestion. The results of pretreatment indicated that sCOD concentrations of hydrolysate were highest (8,233 mg/l) at the fifth day. XDC-2 efficiently degraded the corn stalk by nearly 45%, decreasing the cellulose content by 22.7% and the hemicellulose content by 74.1%. Total levels of volatile products peaked on the fifth day. The six major compounds present were ethanol (0.29 g/l), acetic acid (0.55 g/l), 1,2-ethanediol (0.49 g/l), propionic acid (0.15 g/l), butyric acid (0.22 g/l), and glycerine (2.48 g/l). The results of anaerobic digestion showed that corn stalks treated by XDC-2 produced 68.3% more total biogas and 87.9% more total methane than untreated controls. The technical digestion time for the treated corn stalks was 35.7% shorter than without treatment. The composite microbial system pretreatment could be a cost-effective and environmentally friendly microbial method for efficient biological conversion of corn stalk into bioenergy.     

Reconstruction of lignin and hemicelluloses by aqueous ethanol anti‐solvents to improve the ionic liquid‐acid pretreatment performance of Arundo donax Linn

Ionic liquid (IL)‐acid pretreatment is known to not only enhance the enzymatic hydrolysis efficiency of lignocellulose but also to generate deposits on the surface of fiber by conventional water regeneration, which retard the increment. In this study, ethanol aqueous solution regeneration was developed as a new method to change the substrates characteristics for IL‐acid pretreatment and their effects on the enzymatic hydrolysis were evaluated. Following the IL‐acid reaction, the biomass slurry was subjected to ethanol aqueous solution at various concentration. Results indicated that anti‐solvent choice significantly influenced the reconstruction of both hemicelluloses and lignin as a result of the competition between water and ethanol. The partial removal of hemicelluloses and suitable lignin re‐localization contributed to a more porous structure. Consequently, the cellulose digestibility of aqueous ethanol regenerated samples was dramatically enhanced to ～100% and approximately 11‐ and 2‐fold higher than that of untreated and conventional water regenerated pretreated samples, respectively. A giant leap in the initial rate of enzymatic hydrolysis was also detected in 50% ethanol aqueous solution regenerated samples and only about 10 hr was needed to convert 80% of cellulose to glucose due to the appearance of cellulose II hydrate‐like and more porous structure.     

Visualization of biomass solubilization and cellulose regeneration during ionic liquid pretreatment of switchgrass

Auto‐fluorescent mapping of plant cell walls was used to visualize cellulose and lignin in pristine switchgrass (Panicum virgatum) stems to determine the mechanisms of biomass dissolution during ionic liquid pretreatment. The addition of ground switchgrass to the ionic liquid 1‐n‐ethyl‐3‐methylimidazolium acetate resulted in the disruption and solubilization of the plant cell wall at mild temperatures. Swelling of the plant cell wall, attributed to disruption of inter‐ and intramolecular hydrogen bonding between cellulose fibrils and lignin, followed by complete dissolution of biomass, was observed without using imaging techniques that require staining, embedding, and processing of biomass. Subsequent cellulose regeneration via the addition of an anti‐solvent, such as water, was observed in situ and provided direct evidence of significant rejection of lignin from the recovered polysaccharides. This observation was confirmed by chemical analysis of the regenerated cellulose. In comparison to untreated biomass, ionic liquid pretreated biomass produces cellulose that is efficiently hydrolyzed with commercial cellulase cocktail with high sugar yields over a relatively short time interval. Biotechnol. Bioeng. 2009; 104: 68–75 Published 2009 Wiley Periodicals, Inc.     

Enhancing anaerobic biogasification of corn stover through wet state NaOH pretreatment

A new method of wet state (WS) sodium hydroxide (NaOH) was advanced to pretreat corn stover for enhancing biogas production. The results showed that 88% moisture content, 3-day treatment time and ambient temperature (20 °C) was appropriate for WS NaOH pretreatment. The NaOH dose of 2% and the loading rate of 65 g/L were found to be optimal in terms of 72.9% more total biogas production, 73.4% more methane yield, and 34.6% shorter technical digestion time, as compared to the untreated one. WS pretreatment used 86% shorter treatment time and 66.7% less NaOH dose than solid state one. The analyses of chemical compositions and chemical structures showed that 9.3–19.1% reduction of the contents of total Lignin, cellulose, and hemicellulose (LCH), and 27.1–77.1% increase of hot-water extractives contributed to the enhancement of biogas production. WS NaOH pretreatment could be one of cost-effective methods for high efficient biological conversion of corn stover into bioenergy.     

Correlation between anatomical characteristics of ethanol organosolv pretreated Buddleja davidii and its enzymatic conversion to glucose

Buddleja davidii is a unique biomass that has many attractive agroenergy features, especially its wide range of growth habitat. The anatomical characteristics of B. davidii were investigated before and after ethanol organosolv pretreatment (one of the leading pretreatment technologies) in order to further understand the alterations that occur to the cellular structure of the biomass which can then be correlated with its enzymatic digestibility. Results showed that the ethanol organosolv pretreatment of B. davidii selectively removes lignin from the middle lamella (ML), which does not significantly disrupt the crystalline structure of cellulose. The removal of ML lignin is a major factor in enhancing enzymatic cellulose‐to‐glucose hydrolysis. The pretreatment also causes cell deformation, resulting in cracks and breaks in the cell wall. These observations, together with characterization analysis of the cell wall polymer material, lend support to the hypothesis that the physical distribution of lignin in the biomass matrix is an important structural feature affecting biomass enzymatic digestibility. Biotechnol. Bioeng. 2010;107: 795–801. © 2010 Wiley Periodicals, Inc.     

Enhanced enzymatic hydrolysis of sugarcane bagasse by N-methylmorpholine-N-oxide pretreatment

The cellulose dissolution solvent used in Lyocell process for cellulose fiber preparation, N-methylmorpholine-N-oxide (NMMO) monohydrate, was demonstrated to be an effective agent for sugarcane bagasse pretreatment. Bagasse of 20wt% was readily dissolved in NMMO monohydrate at 130 degrees C within 1h. After dissolution, bagasse could be regenerated by rapid precipitation with water as a porous and amorphous mixture of its original components. The regenerated bagasse exhibited a significant enhancement on enzymatic hydrolysis kinetic. Not only the reducing sugars releasing rate but also hydrolysis yield was enhanced at least twofold as compared with that of untreated bagasse. The cellulose fraction of regenerated bagasse was nearly hydrolyzed to glucose after 72h hydrolysis with Cellulase AP3. The recycled NMMO demonstrated the same performance as the fresh one on bagasse pretreatment for hydrolysis enhancement. The regenerated bagasse was directly used in simultaneous saccharification and fermentation (SSF) for ethanol production by Zymomonas mobilis. No negative effect on ethanol fermentation was observed and ethanol yield approximately 0.15 g ethanol/g baggasse was achieved.     

Fermentability of the hemicellulose‐derived sugars from steam‐exploded softwood (douglas fir)

Steam explosion ofDouglas fir wood chips under low‐severity conditions (log Ro = 3.08 corresponding to 175°C, 7.5 min, and 4.5% SO₂) resulted in the recovery of around 87% of the original hemicellulose component in the water‐soluble stream. More than 80% of the recovered hemicellulose was in a monomeric form. As the pretreatment severity increased from 3.08 to 3.76, hemicellulose recovery dropped to 43% of the original hemicellulose found in Douglas fir chips while the concentration of glucose originating from cellulose hydrolysis increased along with the concentration of sugar degradation products such as furfural and hydroxymethylfurfural. Despite containing a higher concentration of hexose monomers (mainly glucose originating from cellulose degradation), the water‐soluble fraction prepared under high‐severity conditions (log Ro = 3.73 corresponding to 215°C, 2.38 min, and 2.38% SO₂) was not readily fermented. Only the two hydrolyzates obtained at low and medium (195°C, 4.5 min, and 4.5% SO₂) severities were fermented to ethanol using a spent sulfur liquor adapted strain of Saccharomyces cerevisiae. High ethanol yields were obtained for these two hydrolyzates with 0.44 g of ethanol produced per gram of hexose utilized (86% of theoretical). However, the best results of hemicellulose recovery and fermentability were obtained for the low‐severity water‐soluble fraction which was fermented significantly faster than the fraction obtained after medium‐severity treatment probably because it contained higher amounts of fermentation inhibitors. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 284–289, 1999.     

The effects of phosphoric acid pretreatment on conversion of cellulose to ethanol at 45°C using the thermotolerant yeast Kluyveromyces marxianus IMB3

Summary Here we report on the effects of phosphoric acid pretreated cellulose as a substrate for ethanol production by K. marxianus IMB3 using simultaneous saccharification and fermentation systems at 45°C. With untreated, milled filter paper as substrate the maximum amount of ethanol produced was 25% of the maximum theoretical yield. After pre-treatment with 100% phosphoric acid, the yield increased to 42% of the maximum theoretical yield. When untreated microcrystalline cellulose was used as the fermentation substrate, yields of ethanol as 45°C amounted to 16% of the maximum theoretical yield whereas pretreatment of the substrate with phosphoric acid resulted in an increase in ethanol production to 69% of the maximum theoretical yield. This suggests that pretreatment of substrate with phosphoric acid would contribute to a reduction in the amount of exogenous enzyme needed.     

Increase of methane formation by ethanol addition during continuous fermentation of biogas sludge

Very recently, it was shown that the addition of acetate or ethanol led to enhanced biogas formation rates during an observation period of 24 h. To determine if increased methane production rates due to ethanol addition can be maintained over longer time periods, continuous reactors filled with biogas sludge were developed which were fed with the same substrates as the full-scale reactor from which the sludge was derived. These reactors are well reflected conditions of a full-scale biogas plant during a period of 14 days. When the fermenters were pulsed with 50–100 mM ethanol, biomethanation increased by 50–150 %, depending on the composition of the biogas sludge. It was also possible to increase methane formation significantly when 10–20 mM pure ethanol or ethanolic solutions (e.g. beer) were added daily. In summary, the experiments revealed that “normal” methane production continued to take place, but ethanol led to production of additional methane.     

Impact of pretreatment on solid state anaerobic digestion of yard waste for biogas production

Solid state anaerobic digestion, as a safe and environment-friendly technology to dispose municipal solid wastes, can produce methane and reduce the volume of wastes. In order to raise the digestion efficiency, this study investigated the pretreatment of yard waste by thermal or chemical method to break down the complex lignocellulosic structure. The composition and structure of pretreated yard waste were analyzed and characterized. The results showed that the pretreatment decreased the content of cellulose and hemicelluloses in yard waste and in turn improved the hydrolysis and methanogenic processes. The thermal pretreatment sample (P1) had the highest methane yield, by increasing 88 % in comparison with digesting the raw material. The maximum biogas production reached 253 mL/g volatile solids (VS). The largest substrate mass reduction was obtained by the alkaline pretreatment (P5). The VS of the alkaline-treated sample decreased about 60 % in comparison with the raw material.     

Comparative performance of enzymatic and combined alkaline-enzymatic pretreatments on methane production from ensiled sorghum forage

This study investigated the effect of enzymatic and combined alkaline-enzymatic pretreatments on chemical composition and methane production from ensiled sorghum forage. Four commercial enzymatic preparations were tested and the two yielding the highest sugars release were added to evaluate any hydrolytic effect on both untreated and alkaline pretreated samples. In the combined alkaline-enzymatic pretreatment trials, the highest sugar release was found with Primafast and BGL preparations (added at a final concentration 0.12 and 0.20 mL/g TS, respectively), with a total monomeric content of 12 and 6.5 g/L. Fibre composition analysis confirmed that the combined alkaline-enzymatic pretreatment led to cellulose (up to 32 %) and hemicelluloses (up to 56 %) solubilisation, compared to the enzymatic pretreatment alone. BMP tests were performed on both untreated and pretreated samples, and time courses of methane production were fitted. Both enzymatic and combined alkaline-enzymatic pretreatment led to a methane production increase (304 and 362 mL CH₄/g VS), compared to that of untreated sorghum (265 mL CH₄/g VS), as  +15 and  +37 %, respectively. Moreover, higher specific methane production rates, compared to that of untreated sorghum (20.31 mL CH₄/g VS/d), were obtained by applying the enzymatic and combined alkaline-enzymatic pretreatment (33.94 and 31.65 mL CH₄/g VS/d), respectively.     

The effect of various pre‐treatment methods of chromium leather shavings in continuous biogas production

During leather manufacture, high amounts of chromium shavings, wet by‐products of the leather industry, are produced worldwide. They are stable towards temperatures of up to 110°C and enzymatic degradation, preventing anaerobic digestion in a biogas plant. Hitherto, chromium shavings are not utilized industrially to produce biogas. In order to ease enzymatic degradation, necessary to produce biogas, a previous denaturation of the native structure has to be carried out. In our projects, chromium shavings were pre‐treated thermally and mechanically by extrusion and hydrothermal methods. In previous works, we intensively studied the use of these shavings to produce biogas in batch scale and significant improvement was reached when using pre‐treated shavings. In this work, a scale‐up of the process was performed in a continuous reactor using pre‐treated and untreated chromium shavings to examine the feasibility of the considered method. Measuring different parameters along the anaerobic digestion, namely organic matter, collagen content, and volatile fatty acids content, it was possible to show that a higher methane production can be reached and a higher loading rate can be used when feeding the reactor with pre‐treated shavings instead of untreated chromium shavings, which means a more economical and efficient process in an industrial scenario.     

An effective chemical pretreatment method for lignocellulosic biomass with substituted imidazoles

Lignocellulosic biomass is the most abundant naturally renewable organic resource for biofuel production. Because of its recalcitrance to enzymatic degradation, pretreatment is a crucial step before hydrolysis of the feedstock. A variety of pretreatment methods have been developed and intensively studied to achieve optimal yield without imposing significant adverse impact on the environment. Herein, we present a novel chemical pretreatment method using substituted heterocycles with low temperature and short residence time requirements. 1‐Methylimidazole (MI) is a precursor to some imidazolium‐based ionic liquids. In this study, its potential utilization as a biomass pretreatment agent is being investigated for the first time. At mild conditions, such as 25°C for 5 min at ambient pressure, a substantial increase in the hydrolysis rate throughout the entire course of conversion for cellulose substrate was obtained. Furthermore, the pretreatment effectiveness of MI on both untreated and steam‐exploded lignocellulosic biomass including loblolly pine, switchgrass, and sugarcane bagasse has been studied and MI was found to be an efficient delignifier. Remarkable rate enhancement was also observed for the non‐woody lignocellulosic substrates after a short period of MI pretreatment at ambient conditions. The mechanism of MI pretreatment is explored through analysis of cellulose physical properties including crystallinity index, degree of polymerization, accessibility, and lignin dissolution quantification. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 31:25–34, 2015     

Simultaneous saccharification and fermentation of lime-treated biomass

Simultaneous saccharification and fermentation (SSF) was performed on lime-treated switchgrass and corn stover, and oxidatively lime-treated poplar wood to determine their compatibility with Saccharomyces cerevisiae. Cellulose-derived glucose was extensively utilized by the yeast during SSF. The ethanol yields from pretreated switchgrass, pretreated corn stover, and pretreated-and-washed poplar wood were 72%, 62% and 73% of theoretical, respectively, whereas those from -cellulose were 67 to 91% of theoretical. The lower ethanol yields from treated biomass resulted from lower cellulose digestibilities rather than inhibitors produced by the pretreatment. Oxidative lime pretreatment of poplar wood increased the ethanol yield by a factor of 5.6, from 13% (untreated) to 73% (pretreated-and-washed).     

Influence of lignin on the methanization of lignocellulosic wastes

The effect on anaerobic digestion of reducing the lignin content of vine shoots to 1% (w/w), by treatment with sodium chlorite in an acid medium at 80°C, is reported. The yields of methane obtained were 240 ml of CH₄ g⁻¹ of VS (volatile solids) fed for untreated vine shoots, and 370 ml of CH₄ g⁻¹ of VS fed for treated vine shoots. A mathematical model was used to calculate the kinetic parameters H and μ, and the increased biodegradability of the substrate in which lignin had been removed was confirmed. A study of the mass balances of the process under optimum conditions (temperature = 35°C; loading rate of 1 g litre⁻¹ digester day⁻¹) enabled the percentage of degraded cellulose to be calculated (35·5% for untreated vine shoots, 81·5% for the treated vine shoots), as were the volumes of biogas and methane produced per gram of VS introduced (VS₁) and degraded. The blocking effect of lignin on the methanization process was confirmed.     

Alkali‐based AFEX pretreatment for the conversion of sugarcane bagasse and cane leaf residues to ethanol

Sugarcane is one of the major agricultural crops cultivated in tropical climate regions of the world. Each tonne of raw cane production is associated with the generation of 130 kg dry weight of bagasse after juice extraction and 250 kg dry weight of cane leaf residue postharvest. The annual world production of sugarcane is ～1.6 billion tones, generating 279 MMT tones of biomass residues (bagasse and cane leaf matter) that would be available for cellulosic ethanol production. Here, we investigated the production of cellulosic ethanol from sugar cane bagasse and sugar cane leaf residue using an alkaline pretreatment: ammonia fiber expansion (AFEX). The AFEX pretreatment improved the accessibility of cellulose and hemicelluloses to enzymes during hydrolysis by breaking down the ester linkages and other lignin carbohydrate complex (LCC) bonds and the sugar produced by this process is found to be highly fermentable. The maximum glucan conversion of AFEX pretreated bagasse and cane leaf residue by cellulases was ～85%. Supplementation with hemicellulases during enzymatic hydrolysis improved the xylan conversion up to 95–98%. Xylanase supplementation also contributed to a marginal improvement in the glucan conversion. AFEX‐treated cane leaf residue was found to have a greater enzymatic digestibility compared to AFEX‐treated bagasse. Co‐fermentation of glucose and xylose, produced from high solid loading (6% glucan) hydrolysis of AFEX‐treated bagasse and cane leaf residue, using the recombinant Saccharomyces cerevisiae (424A LNH‐ST) produced 34–36 g/L of ethanol with 92% theoretical yield. These results demonstrate that AFEX pretreatment is a viable process for conversion of bagasse and cane leaf residue into cellulosic ethanol. Biotechnol. Bioeng. 2010;107: 441–450. © 2010 Wiley Periodicals, Inc.