Two-stage dilute-acid pretreatment of softwoods

Whole treechips obtained from softwood forest thinnings were pretreated via single-and two-stage dilute-sulfuric acid pretreatment. Whole-tree chips were impregnated with dilute sulfuric acid and steam treated in a 4-L steam explosion reactor. In single-stage pretreatment, wood chips were treated using a wide range of severity. In two-stage pretreatment, the first stage was carried out at low severity tomaximize hemicellulose recovery. Solubilized sugars were recovered from the first-stage prehydrolysate by washing with water. In the second stage, water-insoluble solids from first-stage prehydrolysate were impregnated with dilute sulfuric acid, then steam treated at more severe conditions to hydrolyze a portion of the remaining cellulose to glucose and to improve the enzyme digestibility. The total sugar yields obtained after enzymatic hydrolysis of two-stage dilute acid-pretreated samples were compared with sugar yields from single-stage pretreatment. The overall sugar yield from two-stage dilute-acid pretreatment was approx 10% higher, and the net enzyme requirement was reduced by about 50%. Simultaneous saccharification and fermentation using an adapted Saccharomyces cerevisiae yeast strain further improved cellulose conversion yield and lowered the enzyme requirement.     

Evaluation of Ethanol Production from Corncob Using Scheffersomyces (Pichia) stipitis CBS 6054 by Volumetric Scale-up

In scale-up, the potential of ethanol production by dilute sulfuric acid pretreatment using corncob was investigated. Pretreatments were performed at 170 °C with various acid concentrations ranging from 0% to 1.656% based on oven dry weight. Following pretreatment, pretreated biomass yield ranged from 59% to 67%. More than 90% of xylan was removed at 0.828% of sulfuric acid. At same pretreatment condition, the highest glucose yield obtained from pretreated biomass by enzymatic hydrolysis was about 76%, based on a glucan content of 37/100 g. In hydrolysate obtained by pretreatment, glucose concentration was low, while xylose concentration was significantly increased above 0.368% of sulfuric acid. At 1.656% of sulfuric acid, xylose and glucose concentration was highest. In subsequent, fermentation with hydrolysate, maximal ethanol yield was attained after 24 h with 0.368% of sulfuric acid. The fermentation efficiency of hydrolysate obtained by enzymatic hydrolysis reached a maximum of 75% at an acid charge of 0.368%.     

Comparison of pretreatment methods on the enzymatic saccharification of aspen wood

Five different chemical pretreatments, using dilute sulfuric acid, sodium hydroxide, hydrogen peroxide and sodium hydroxide, peroxymonosulfate, and acetic acid, were applied to aspen thermomechanical fibers. The pretreated fibers were submitted to enzymatic hydrolysis and the liberated glucose was monitored. High glucose concentrations were observed for the peroxymonosulfate and the acetic acid pretreated samples. Glucose concentrations greater than 25 g/L were obtained in these cases. This corresponds to conversions on the order of 90% of the pretreated substrate glucose content.     

Hybrid process for the conversion of lignocellulosic materials

Because of the recalcitrant nature of lignocellulosic materials, it is important to pretreat the biomass in order to obtain a suitable material for the bioconversion. In this study, two different types of pretreatments were performed. The first experiment used a 2-gal Parr reactor operated at 140, 150,160, and 170‡C with sulfuric acid concentrations varying from 0.5 to 2%. A second pretreatment was performed with a two-stage low-temperature process. The first-stage pretreatment was performed at 100 or 120‡C with sulfuric acid concentrations of 0.5, 2, and 5% followed by a secondstage pretreatment at 120‡C with 2% acid concentration. The best residues for enzymatic hydrolysis and simultaneous saccharification and fermentations (SSF) came from the higher temperature pretreatment with the Parr reactor. However, a large portion of the xylose fraction was degraded to furfural and glucose was degraded to HMF. On the contrary, the two-stage low temperature pretreatment resulted in a very low percentage of xylose degradation, and no glucose degradation. The residues from this two-stage pretreatment performed satisfactorily toward the production of ethanol by SSFs. This study discusses the results obtained from these experiments.     

Xylan hydrolysis in zinc chloride solution

Xylan is the major component of hemicellulose, which consists of up to one-third of the lignocellulosic biomass. When the zinc chloride solution was used as a pretreatment agent to facilitate cellulose hydrolysis, hemicellulose was hydrolyzed during the pretreatment stage. In this study, xylan was used as a model to study the hydrolysis of hemicellulose in zinc chloride solution. The degradation of xylose that is released from xylan was reduced by the formation of zinc-xylose complex. The xylose yield was >90% (w/w) at 70°C. The yield and rate of hydrolysis were a function of temperature and the concentration of zinc chloride. The ratio of zinc chloride can be decreased from 9 to 1.3 (w/w). At this ratio, 76% of xylose yield was obtained. When wheat straw was pretreated with a concentrated zinc chloride solution, the hemicellulose hydrolysate contained only xylose and trace amounts of arabinose and oligosaccharides. With this approach, the hemicellulose hydrolysate can be separated from cellulose residue, which would be hydrolyzed subsequently to glucose by acid or enzymes to produce glucose. This production scheme provided a method to produce glucose and xylose in different streams, which can be fermented in separated fermenters.     

A technical and economic analysis of acid-catalyzed steam explosion and dilute sulfuric acid pretreatments using wheat straw or aspen wood chips

Lignocellulosic biomass is one of the most plentiful and potentially cheapest feedstocks for ethanol production. The cellulose component can be broken down into glucose by enzymes and then converted to ethanol by yeast. However, hydrolysis of cellulose to glucose is difficult, and some form of pretreatment is necessary to increase the susceptibility of cellulose to enzymatic attack. An analysis has been completed of two pretreatment options, dilute sulfuric acid hydrolysis and sulfur dioxide impregnated steam explosion, for two feedstocks, wheat straw and aspen wood chips. Detailed process flow sheets and material and energy balances were used to generate equipment cost information. A technical and economic analysis compared the two feedstocks for each of the two pretreatments. For the same pretreatment, sugars produced from aspen wood hydrolysis were cheaper because of the higher carbohydrate content of aspen, whereas dilute acid pretreatment is favored over acid-catalyzed steam explosion.     

Summary of findings from the Biomass Refining Consortium for Applied Fundamentals and Innovation (CAFI): corn stover pretreatment

The Biomass Refining Consortium for Applied Fundamentals and Innovation, with members from Auburn University, Dartmouth College, Michigan State University, the National Renewable Energy Laboratory, Purdue University, Texas A&M University, the University of British Columbia, and the University of California at Riverside, has developed comparative data on the conversion of corn stover to sugars by several leading pretreatment technologies. These technologies include ammonia fiber expansion pretreatment, ammonia recycle percolation pretreatment, dilute sulfuric acid pretreatment, flowthrough pretreatment (hot water or dilute acid), lime pretreatment, controlled pH hot water pretreatment, and sulfur dioxide steam explosion pretreatment. Over the course of two separate USDA- and DOE-funded projects, these pretreatment technologies were applied to two different corn stover batches, followed by enzymatic hydrolysis of the remaining solids from each pretreatment technology using identical enzyme preparations, enzyme loadings, and enzymatic hydrolysis assays. Identical analytical methods and a consistent material balance methodology were employed to develop comparative sugar yield data for each pretreatment and subsequent enzymatic hydrolysis. Although there were differences in the profiles of sugar release, with the more acidic pretreatments releasing more xylose directly in the pretreatment step than the alkaline pretreatments, the overall glucose and xylose yields (monomers + oligomers) from combined pretreatment and enzymatic hydrolysis process steps were very similar for all of these leading pretreatment technologies. Some of the water-only and alkaline pretreatment technologies resulted in significant amounts of residual xylose oligomers still remaining after enzymatic hydrolysis that may require specialized enzyme preparations to fully convert xylose oligomers to monomers.     

Evaluation of different biomass materials as feedstock for fermentable sugar production

Saline crops and autoclaved municipal organic solid wastes were evaluated for their potential to be used as feedstock for fermentable sugar production through dilute acid pretreatment and enzymatic hydrolysis. The saline crops included two woods, athel (Tamarix aphylla L) and eucalyptus (Eucalyptus camaldulensis), and two grasses, Jose tall wheatgrass (Agropyron elongatum), and creeping wild rye (Leymus triticoides). Each of the biomass materials was first treated with dilute sulfuric acid under selected conditions (acid concentration =1.4% (w/w), temperature =165 degrees C, and time =8 min) and then treated with the enzymes (cellulases and beta-glucosidase). The chemical composition (cellulose, hemicellulose, and lignin contents) of each biomass material and the yield of total and different types of sugars after the acid and enzyme treatment were determined. The results showed that among the saline crops evaluated, the two grasses (creeping wild rye and Jose tall wheatgrass) had the highest glucose yield (87% of total cellulose hydrolyzed) and fastest reaction rate during the enzyme treatment. The autoclaved municipal organic solid wastes showed reasonable glucose yield (64%). Of the two wood species evaluated, Athel has higher glucose yield (60% conversion of cellulose) than eucalyptus (38% conversion of cellulose).     

Acid recovery by electrodialysis and its economic implications for concentrated acid hydrolysis of wood

Satisfactory separation of either hydrochloric or sulfuric acid from sugars in wood hydrolyzates by application of membrane technology is technically feasible. The permeability of disaccharides is less than 1% that of the acids. Acid flux in diffusion dialysis is only 6% of acid flux at optimum current density in electrodialysis. Critical parameters for economic feasibility are acid to wood ratio in hydrolysis, current efficiency, and membrane service life. Best case estimates project total costs for sulfuric acid recovery and loss of about $0.02 per pound of glucose produced.     

The effect of agitation speed,enzyme loading and substrate concentration on enzymatic hydrolysis of cellulose from brewer’s spent grain

Brewer’s spent grain components (cellulose, hemicellulose and lignin) were fractionated in a two-step chemical pretreatment process using dilute sulfuric acid and sodium hydroxide solutions. The cellulose pulp produced was hydrolyzed with a cellulolytic complex, Celluclast 1.5 L, at 45 °C to convert the cellulose into glucose. Several conditions were examined: agitation speed (100, 150 and 200 rpm), enzyme loading (5, 25 and 45 FPU/g substrate), and substrate concentration (2, 5 and 8% w/v), according to a 2³ full factorial design aiming to maximize the glucose yield. The obtained results were interpreted by analysis of variance and response surface methodology. The optimal conditions for enzymatic hydrolysis of brewer’s spent grain were identified as 100 rpm, 45 FPU/g and 2% w/v substrate. Under these conditions, a glucose yield of 93.1% and a cellulose conversion (into glucose and cellobiose) of 99.4% was achieved. The easiness of glucose release from BSG makes this substrate a raw material with great potential to be used in bioconversion processes.     

Modeling the enzymatic hydrolysis of dilute-acid pretreated Douglas Fir

Glucose yield from the enzymatic hydrolysis of cellulose was investigated as a function of cellulase enzyme loading (7–36 filter paper units [FPU]/g cellulose) and solids concentration (7–18% total solids) for up to 72 h on dilute sulfuric-acid pretreated Douglas Fir. The saccharification was performed on whole hydrolysate with no separation or washing of the solids. Enzyme loading had a significant effect on glucose yield; solids concentration had a much smaller effect even at higher glucose concentrations. The data were used to generate an empirical model for glucose yield, and to fit parameters of a cellulose hydrolysis kinetic model. Both models could be used for economic evaluation of a separate hydrolysis and fermentation process.     

Initial design of a dilute sulfuric acid pretreatment process for aspen wood chips

A preliminary process design for dilute sulfuric acid pretreatment of aspen wood chips in order to obtain fermentable sugars has been prepared and subjected to an economic evaluation. The process design was prepared according to experimental data on the kinetics of dilute sulfuric acid prehydrolysis and particle size effects obtained in this study and our previous work. The initial economic evaluation shows woodchips are 56% of the cost of production, whereas the reactor is only 4%, and the comminution operation is just under 10%, indicating that the process economics are extremely vulnerable to feedstock costs and are thus yield-sensitive. Although chances for major cost improvements by modification of the reactor design and finding alternatives to dry milling of aspen chips to small (20–80 mesh) particles needed for acid penetration and enzymatic saccharification are not great, design improvements of the process will necessitate development of a cheaper acid resistant pretreatment reactor and a less energy intensive comminution system. Experimental results on effects of particle size on the dilute acid pretreatment design are presented.     

Lactic acid production from cellulosic material by synergetic hydrolysis and fermentation

The hydrolysis process on corncob residue was catalyzed synergetically by the cellulase from Trichoderma reesei and the immobilized cellobiase. The feedback inhibition to cellulase reaction caused by the accumulation of cellobiose was eliminated efficiently. The hydrolysis yield of corncob residue was 82.5%, and the percentage of glucose in the reducing sugar reached 88.2%. The glucose in the cellulosic hydrolysate could be converted into lactic acid effectively by the immobilized cells of Lactobacillus delbrueckii. When the enzymatic hydrolysis of cellulose and the fermentation of lactic acid were coupled together, no glucose was accumulated in the reaction system, and the feedback inhibition caused by glucose was also eliminated. Under the batch process of synergetic hydrolysis and lactic acid fermentation with 100 g/L of cellulosic substrate, the conversion efficiency of lactic acid from cellulose and the productivity of lactic acid reached 92.4% and 0.938 g/(L·h), respectively. By using a fed-batch technique, the total concentration of cellulosic substrate and lactic acid in the synergetic process increased to 200 and 107.5 g/L, respectively, whereas the dosage of cellulase reduced from 20 to 15 IU/g of substrate in the batch process. The results of the bioconversion of renewable cellulosic resources were significant.     

Tailoring Wet Explosion Process Parameters for the Pretreatment of Cocksfoot Grass for High Sugar Yields

The pretreatment of lignocellulosic biomass is crucial for efficient subsequent enzymatic hydrolysis and ethanol fermentation. In this study, wet explosion (WEx) pretreatment was applied to cocksfoot grass and pretreatment conditions were tailored for maximizing the sugar yields using response surface methodology. The WEx process parameters studied were temperature (160–210 °C), retention time (5–20 min), and dilute sulfuric acid concentration (0.2–0.5 %). The pretreatment parameter set E, applying 210 °C for 5 min and 0.5 % dilute sulfuric acid, was found most suitable for achieving a high glucose release with low formation of by-products. Under these conditions, the cellulose and hemicellulose sugar recovery was 94 % and 70 %, respectively. The efficiency of the enzymatic hydrolysis of cellulose under these conditions was 91 %. On the other hand, the release of pentose sugars was higher when applying less severe pretreatment conditions C (160 °C, 5 min, 0.2 % dilute sulfuric acid). Therefore, the choice of the most suitable pretreatment conditions is depending on the main target product, i.e., hexose or pentose sugars.     

Potential of Agricultural Residues and Hay for Bioethanol Production

Production of bioethanol from agricultural residues and hays (wheat, barley, and triticale straws, and barley, triticale, pearl millet, and sweet sorghum hays) through a series of chemical pretreatment, enzymatic hydrolysis, and fermentation processes was investigated in this study. Composition analysis suggested that the agricultural straws and hays studied contained approximately 28.62-38.58% glucan, 11.19-20.78% xylan, and 22.01-27.57% lignin, making them good candidates for bioethanol production. Chemical pretreatment with sulfuric acid or sodium hydroxide at concentrations of 0.5, 1.0, and 2.0% indicated that concentration and treatment agent play a significant role during pretreatment. After 2.0% sulfuric acid pretreatment at 121 degrees C/15 psi for 60 min, 78.10-81.27% of the xylan in untreated feedstocks was solubilized, while 75.09-84.52% of the lignin was reduced after 2.0% sodium hydroxide pretreatment under similar conditions. Enzymatic hydrolysis of chemically pretreated (2.0% NaOH or H2SO4) solids with Celluclast 1.5 L-Novozym 188 (cellobiase) enzyme combination resulted in equal or higher glucan and xylan conversion than with Spezyme(R) CP- xylanase combination. The glucan and xylan conversions during hydrolysis with Celluclast 1.5 L-cellobiase at 40 FPU/g glucan were 78.09 to 100.36% and 74.03 to 84.89%, respectively. Increasing the enzyme loading from 40 to 60 FPU/g glucan did not significantly increase sugar yield. The ethanol yield after fermentation of the hydrolyzate from different feedstocks with Saccharomyces cerevisiae ranged from 0.27 to 0.34 g/g glucose or 52.00-65.82% of the theoretical maximum ethanol yield.     

Dilute-acid hydrolysis of sugarcane bagasse at varying conditions

Sugarcane bagasse, a byproduct of the cane sugar industry, is an abundant source of hemicellulose that could be hydrolyzed to yield a fermentation feedstock for the production of fuel ethanol and chemicals. The effects of sulfuric acid concentration, temperature, time, and dry matter concentration on hemicellulose hydrolysis were studied with a 20-L batch hydrolysis reactor using a statistical experimental design. Even at less severe conditions considerable amounts (>29%) of the hemicellulose fraction could be extracted. The percentage of soluble oligosaccharides becomes very low in experiments with high yields in monosaccharides, which indicates that the cellulose fraction is only slightly affected. For the sugar yields, acid concentration appears to be the most important parameter, while for the formation of sugar degradation products, temperature shows the highest impact. It could be demonstrated that the dry matter concentration in the reaction slurry has a negative effect on the xylose yield that can be compensated by higher concentrations of sulfuric acid owing to a positive interaction between acid concentration and dry matter contents.     

Production of reducing sugar from corn stover by dilute acid hydrolysis co-catalyzed with metal salts under microwave radiation

Acid hydrolysis of corn stover under microwave radiation by adding four metal salt co-catalysts (ferrous sulfate, copper sulfate, zinc sulfate, and manganese sulfate) to produce reducing sugar was investigated. The influences of microwave power, reaction time, sulfuric acid concentration, and co-catalyst concentration on the yield of reducing sugar were studied. The results show that the catalytic effects of ferrous sulfite and copper sulfate are more obvious for the yield of reducing sugar compared with only dilute acid hydrolysis with microwave radiation. Long-time microwave irradiation resulted in secondary hydrolysis, generating small-molecule compounds and decreasing the yield of reducing sugar. The reducing sugar yield was increased by decreasing the hydrolysis reaction time and microwave radiation power. In terms of reducing sugar yield, the four kinds of metal salt co-catalyst act in the order: ferrous sulfate > copper sulfate > sulfuric acid zinc sulfate > manganese sulfate. At concentration of 3.0 %, the best was ferrous sulfate, producing yield of 35.10 %.     

Application of a depolymerization model for predicting thermochemical hydrolysis of hemicellulose

Literature data were collected and analyzed to guide selection of conditions for pretreatment by dilute acid and water-only hemicellulose hydrolysis, and the severity parameter was used to relate performance of different studies on a consistent basis and define attractive operating conditions. Experiments were then run to confirm performance with corn stover. Although substantially better hemicellulose sugar yields are observed when acid is added, costs would be reduced and processing operations simplified if less acid could be used while maintaining good yields, and understanding the relationship between operating conditions and yields would be invaluable to realizing this goal. However, existing models seldom include the oligomeric intermediates prevalent at lower acid levels, and the few studies that include such species do not account for the distribution of chain lengths during reaction. Therefore, the polymeric nature of hemicellulose was integrated into a kinetic model often used to describe the decomposition of synthetic polymers with the assumption that hemicellulose linkages are randomly broken during hydrolysis. Predictions of monomer yields were generally consistent with our pretreatment data, data reported in the literature, and predictions of other models, but the model tended to overpredict oligomer yields. These differences need to be resolved by gathering additional data and improving the model.     

Continuous countercurrent extraction of hemicellulose from pretreated wood residues

Two-stage dilute acid pretreatment followed by enzymatic cellulose hydrolysisis an effectivemethod for obtaining high sugar yields from wood residuessuchassoftwood forest thinnings. In the first-stage hydrolysis step, most of the hemicellulose is solubilized using relatively mild conditions. The soluble hemicellu losic sugars are recovered from the hydrolysateslurry by washing with water. The washed solids are then subjected tomoresevere hydrolysis conditions to hydrolyze approx 50% of the cellulose to glucose. The remaining cellulose can further be hydrolyzed with cellulase enzyme. Our process simulation indicates that the amount of water used in the hemicellulose recovery step has a significan tim pact on the cost of ethanol production. It is important to keep water usage as low as possible while mainta ining relatively high recovery of solublesugars. To achieve this objective, a prototype pilot-scale continuous countercurrent screw extractor was evaluated for the recovery of hemicellulose from pretreated forest thinnings. Using the 274-cm (9-ft) long extractor, solubles recoveries of 98, 91, and 77% were obtained with liquid-to-insoluble solids (L/1S) ratios of 5.6, 3.4, and 2.1, respectively. An empirical equation was developed to predict the performance of the screwextractor. This equation predicts that soluble sugar recovery above 95% can be obtained with an L/IS ratio as low as 3.0.     

Severity function describing the hydrolysis of xylan using carbonic acid

Beech wood derived xylan to hydrolyzed to predominantly xylose monomer units after exposure to hot, compressed liquid water saturated with carbon dioxide. Similar treatment without CO₂ saturation resulted in only minor hydrolysis and a smaller fraction of monomers among the hydrolysis products. Severity of the hydrolysis reaction was correlated to reaction time, temperature, and carbon dioxide partial pressure and followed a function similar to those used to characterize mineral acid systems. Results from parallel hydrolysis experiments with an aqueous system and a very dilute sulfuric acid system allowed an approximation of the dissociation constant of carbonic acid in the temperature range of 170–230°C. Results suggest that carbonic acid may be a viable reagent for promoting hydrolysis without mineral acids, especially in the case of a bioprocessing plant that produces carbon dioxide.