Utilization of pretreated bagasse for the sustainable bioproduction of cellulase by Aspergillus nidulans MTCC344 using response surface methodology

Response surface methodology (RSM) was used to optimize the conditions for the production of endo β-1,4 glucanase, a component of cellulase by Aspergillus nidulans MTCC344 under solid state fermentation, using bagasse as the chief substrate. A four-factor-five-level central composite design was employed for experimental design and analysis of the results. Maximum cellulase activity (CMCase was 28.96 U g⁻¹) can be attained at the optimum conditions, 16.8 mm bagasse bed height, 60% moisture content, pH 4.25 and temperature 40 °C in the solid state fermenter. These data were rather close to the experimental results obtained (CMCase was 28.84 U g⁻¹). A. nidulans MTCC344 was able to hydrolyze pretreated bagasse completely after 8 days of incubation with significant endo β-1,4 glucanase activities. The results of Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) of bagasse showed structural changes through pretreatment, in favor of enzymatic hydrolysis. Bagasse with alkali pretreatment using sodium hydroxide is a source of lignocelluloses able to improve the yield of endo β-1,4 glucanase by the strain of A. nidulans. The endo β-1,4 glucanase produced during the bioconversion of cellulose to glucose by A. nidulans MTCC344 is strongly dependent on the pretreatment given before hydrolysis.     

Process optimization for ethanol production from photoperiod-sensitive sorghum: Focus on cellulose conversion

Photoperiod-sensitive sorghum, as a competitive biomass for ethanol production, was investigated to develop an integrated process for improving ethanol yield. Response surface methodology was employed to study the relationship between pretreatment variables (including temperature, sulfuric acid concentration, and reaction time) and cellulose recovery, as well as efficiency of enzymatic hydrolysis (EEH) in the solid part. Recovery yield decreased and EEH increased as the pretreatment temperature, acidic concentration, and reaction time increased. A model was successfully developed to predict total glucose yield with a maximum value of 82.2%. Conditions of co-fermentation were also optimized, and the optimal ethanol yield was obtained with constant-temperature simultaneous saccharification and fermentation at 38 °C. Acetate buffer at a concentration of 50 mM was found helpful for increasing efficiency of enzymatic hydrolysis, as well as ethanol yield. The maximum ethanol yield was 0.21 g ethanol per dry mass at the conditions of 38 °C, 0.05 g yeast/L, and 50 mM acetate buffer. A complete cellulose balance was provided for the whole process.     

Effects of washing, milling and loading enzymes on the enzymatic hydrolysis of a steam pretreated sugarcane bagasse

Enzymatic hydrolysis of steam pretreated sugarcane bagasse was performed to investigate the production of glucose and xylose. A blend of industrial enzymes (Novozymes) was used. The enzymes were used either without having been washed or having been washed with water or 1% aqueous NaOH solution. The influence of the size of sugarcane bagasse and the proportion of the enzymatic cocktail, which was composed of endoglucanases/cellobiohydrolases (Celluclast 1.5L) and β-glucosidase (Novozym 188) cellulases mixtures, was investigated. The tests were performed at a temperature of 50 °C and at a pH of 4.8 during a period of 72 h. The assays that were conducted with a pretreated sugarcane bagasse that was washed with a 1% aqueous NaOH solution without milling led to the largest amounts of glucose after 72 h, which was independent of the proportions of enzymes used in experiments with the smallest amounts of enzymes. The largest amount of xylose was obtained with a pretreated sugarcane bagasse that was not washed.     

Ethanol production from alkali- and ozone-treated cotton stalks using thermotolerant Pichia kudriavzevii HOP-1

In the present study, milled cotton stalks were subjected to alkali pretreatment with NaOH at 1-4% (w/v) concentrations at 121 °C for time ranging from 30 to 90 min. Ozone pretreatment was performed by passing 45 mg/L of ozone gas over 2 mm cotton stalks for 150 min at a flow rate of 0.37 L/min. The residual biomass from 4% alkali pretreatment for 60 min showed 46.6% lignin degradation accompanied by 83.2% increase in glucan content, compared with the untreated biomass. Hydrolysis of 4% alkali-treated and ozone-treated cotton stalks was conducted using enzyme combination of 20 filter paper cellulase units/gram dried substrate (FPU/g-ds), 45 IU/g-ds β-glucosidase and 15 IU/g-ds pectinase. Enzymatic hydrolysis of alkali-treated and ozone-treated biomass after 48 h resulted in 42.29 g/L glucose, 6.82 g/L xylose and 24.13 g/L glucose, 8.3 g/L xylose, respectively. About 99% of glucose was consumed in 24 h by Pichia kudriavzevii HOP-1 cells resulting in 19.82 g/L of ethanol from alkali-treated cotton stalks and 10.96 g/L of ethanol from ozone-treated cotton stalks. Simultaneous saccharification and fermentation of the alkali-treated cotton stalks after 12-h pre-hydrolysis resulted in ethanol concentration, ethanol yield on dry biomass basis and ethanol productivity of 19.48 g/L, 0.21 g/g and 0.41 g/L/h, respectively which holds promise for further scale-up studies. To the best of our knowledge, this is the first study employing SSF for ethanol production from cotton stalks.     

Optimization of ethanol production by Saccharomyces cerevisiae UFPEDA 1238 in simultaneous saccharification and fermentation of delignified sugarcane bagasse

Ethanol production by Saccharomyces cerevisiae UFPEDA1238 was performed in simultaneous saccharification and fermentation of delignified sugarcane bagasse. Temperature (32 °C, 37 °C), agitation (80; 100 rpm), enzymatic load (20 FPU/g cellulose and 10%, v/v β-glucosidase or 10 FPU/g cellulose and 5% β-glucosidase) and composition of culture medium were evaluated. Ethanol concentration, enzymatic convertibility of cellulose and volumetric productivity were higher than 25 g/L, 72% and 0.70 g/L h, respectively, after 30 h, when the culture medium 1 and 20 FPU/g cellulose/10%, v/v β-glucosidase or the culture medium 2 and 10 FPU/g cellulose/5% β-glucosidase were used in SSF at 37 °C and 80 rpm. In the SSF with culture medium 2 (supplemented with ammonium, phosphate, potassium and magnesium), 150 L ethanol/t bagasse was achieved, with minimum enzyme loading (10 FPU/g cellulose and 5%, v/v β-glucosidase) for 8%, w/v of solids, which is often an important requirement to provide cost-efficient second generation ethanol processes.     

Steam explosion pretreatment reproduction and alkaline delignification reactions performed on a pilot scale with sugarcane bagasse for bioethanol production

This work was focused on the steam explosion pretreatment reproduction and alkaline delignification reactions on a pilot scale for the ethanol production, through different varieties of natural sugarcane bagasse, pretreated bagasse and delignified pretreated bagasse (cellulosic pulp). The possible chemical composition differences of the various types of bagasse, as well as the chemical composition variations of the materials in the 20 processes of pretreatment and delignification on the pilot scale were verified. The analytical results of the 20 samples of most diverse varieties and origins of natural sugarcane bagasse considering planting soils, planting periods and weather; show no significant chemical differences. It is evident that only with the chemical composition it is not possible to verify the differences between the varieties of sugarcane bagasses. The research results may offer some evidences of these varieties, but it is not a reliable parameter. The pilot process of steam explosion pretreatment and the alkaline delignification process of pretreated material showed through analytical results a good capacity of reproduction, as the standard differences were below 2.7. The average allowed in the pretreatment and alkaline delignification processes were 66.1 ± 0.8 and 51.5 ± 2.6 respectively, ensuring an excellent reproduction capacity of the processes obtained through chemical characterizations.     

Guayule as a feedstock for lignocellulosic biorefineries using ammonia fiber expansion (AFEX) pretreatment

Natural rubber latex extraction from guayule leaves behind greater than 90% (by weight) of agricultural residue as a feedstock suitable for conversion to biofuels via a thermochemical or biochemical route. Untreated guayule shrub and bagasse (after latex extraction) has shown to be very recalcitrant to enzymatic hydrolysis, necessitating application of a chemical pretreatment to enhance cellulase accessibility. The objective of this work was to carry out detailed compositional analysis, ammonia fiber expansion (AFEX¹) pretreatment, enzymatic hydrolysis and ethanol fermentation for various guayule-derived biomass fractions. Plant feedstocks tested were derived from two sources; (a) a mature 2007 AZ-2 whole guayule shrub plant obtained from USDA/ARS² research fields, and (b) the guayule latex-extracted commercial grade bagasse (62505) from Yulex Corporation. Compositional analysis and enzymatic hydrolysis were carried out using standard NREL³ protocols (www.nrel.gov/biomass/analytical_procedures.html). AFEX pretreatment was carried out using concentrated ammonium hydroxide at elevated temperatures for desired residence times in a pressurized reactor. Yeast fermentations on biomass hydrolyzates were carried out micro-aerobically using Saccharomyces cerevisiae (424A strain) in shake flasks.AFEX pretreatment was found to substantially improve overall enzymatic digestibility by 4-20 fold for both untreated guayule shrub and latex-extracted bagasse. Maximum glucan and xylan conversion achieved for the latex-extracted bagasse was 40% and 50%, respectively. The yeast was readily able to ferment both glucose and xylose to ethanol from the guayule bagasse hydrolyzate with or without external nutrient supplementation (i.e., yeast extract and tryptone). Our results highlight the possible utilization of guayule as a feedstock for lignocellulosic refineries co-producing natural rubber latex and biofuels. However, further process improvements (e.g., lignin/resin extraction and cellulose decrystallization using a modified AFEX process) are necessary to increase the effectiveness of ammonia-based pretreatments for further enhancing enzymatic digestibility of guayule-derived hardwood biomass.     

Optimization of formic/acetic acid delignification of Miscanthus × giganteus for enzymatic hydrolysis using response surface methodology

A Box-Behnken experimental design and response surface methodology were employed to optimize the pretreatment parameters of a formic/acetic acid delignification treatment of Miscanthus × giganteus for enzymatic hydrolysis. The effects of three independent variables, namely cooking time (1, 2 and 3 h), formic acid/acetic acid/water ratio (20/60/20, 30/50/20 and 40/40/20) and temperature (80, 90 and 107 °C) on pulp yield, residual Klason lignin content, concentration of degradation products (furfural and hydroxymethylfurfural) in the black liquor, and enzymatic digestibility of the pulps were investigated. The major parameter influencing was the temperature for pulp yield, delignification degree, furfural production and enzymatic digestibility. According to the response surface analysis the optimum conditions predicted for a maximum enzymatic digestibility of the glucan (75.3%) would be obtained using a cooking time of 3 h, at 107 °C and with a formic acid/acetic acid/water ratio of 40/40/20%. Glucan digestibility was highly dependent on the delignification degree.     

Improvement of biogas production from oil palm empty fruit bunches (OPEFB)

Oil palm empty fruit bunches (OPEFB), a waste lignocellulosic material, which is the main byproduct of vegetable oil production industries in Indonesia and Malaysia, was utilized as a source for biogas production. Pretreatments using NaOH as well as phosphoric acid were investigated to improve the biogas production. Clear positive effects of the pretreatments on the yield of methane were observed. The best improvement was achieved when 8% NaOH for 60 min was used for the pretreatment, which resulted in 100% improvement in the yield of methane production. In addition, treatment with phosphoric acid resulted in 40% improvement in the methane yield compared with that of the untreated material. The results showed that the carbohydrate content of OPEFB could be efficiently converted to methane under the anaerobic digestion process. 97% of the theoretical value of methane production was achieved after the pretreatment with NaOH for 60 min. Moreover, the initial rate of methane production was also increased by more than 85% after the treatment with NaOH compared with that of the untreated OPEFB.     

Processing of Lespedeza stalks by pretreatment with low severity steam and post-treatment with alkaline peroxide

The steam pre-treatment with low severity preserves valuable biomass components, and further delignification with alkaline peroxide could improve hydrolysis. A combination of low severity steam pretreatment and alkaline peroxide post-treatment of Lespedeza stalks was investigated. The post-treatment of steam-pretreated Lespedeza stalks with alkaline peroxide significantly increased the cellulose content and changed the structure of the cellulose-rich fractions. A glucose yield of 503.5 mg g⁻¹ raw material from enzyme hydrolysis was obtained when the steam-pretreated material (184 °C for 4 min) was post-treated with 2% hydrogen peroxide at 60 °C for 24 h with a substrate concentration of 3.3%. Its hydrolysis yield is 88.8%, which is higher than that of samples processed by steam pretreatment alone (63.7%). The samples obtained by post-treatment with alkaline peroxide were found to have a smoother surface and looser structure in scanning electron microscopy images. The isolated lignin preparations had a yield range from 10.9 to 14.7 (% dry matter). The lignin was characterized by thermogravimetric analysis/differential thermal analysis, Fourier transform infrared spectroscopy, and gel permeation chromatography. Alkaline peroxide treatment increased the thermal stability of lignin, and decreased the amounts of all functional groups. Depolymerization and repolymerization occurred during the alkaline peroxide treatment.     

Conversion of starch from dry common beans (Phaseolus vulgaris L.) to ethanol

Dry common beans (Phaseolus vulgaris L.) were evaluated for potential conversion of starch to ethanol. Eight varieties of beans with average starch content of 46% (db) were assayed in a laboratory-scaled process based upon the commercial corn dry grind fermentation process. Ethanol yield was 0.43-0.51 g ethanol/g glucose (0.19-0.23 g ethanol/g beans). The average ethanol yield for the eight bean types was 92% of maximum theoretical yield, demonstrating that starch from beans could be efficiently converted to ethanol. Ethanol concentration obtained from 20% (w/w) solids loading was 3.5-4.4% (w/v). The residual fermentation solids contained, on a dry basis, 37.1-43.6% crude protein, 10.8-15.1% acid detergent fiber and 19.1-31.3% neutral detergent fiber.     

Optimization of phosphoric acid catalyzed fractionation and enzymatic digestibility of flax shives

Flax shives are the woody residue left over from processing flax straw into fiber, and are an abundant renewable lignocellulosic material with a potential for the conversion into bioethanol and other value added products. In this study, prior to enzymatic hydrolysis for the liberation of fermentable sugars, such as glucose and xylose, flax shives were treated with concentrated phosphoric acid. In order to optimize the phosphoric acid pretreatment and enzymatic hydrolysis steps, the effects of three process variables on the fractionation of flax shives, and enzymatic digestibility of pretreated flax shives were evaluated. The optimization process employed a central composite design (CCD), where the variables selected were concentration of phosphoric acid (40.8–86.2%), pretreatment time (9.5–110.5 min), and cellulase loading (13.1–71.9 FPU/g cellulose). Using three-variable and five-level CCD, all tested independent variables were identified to have significant effects (P < 0.05) on the digestibility of pretreated flax shives. It was found that the level of phosphoric acid (P < 0.0001) affects the digestibility most significantly when compared with other variables. When the optimization was conducted under a constrain of minimum cellulase loading, the maximum digestibility of 94.8% was predicted when the phosphoric acid concentration, pretreatment time, and cellulase loading were 86.2%, 110.5 min, and 13.1 FPU/g cellulose at 50 °C and 120 h, respectively. Under these conditions, digestibility of pretreated flax shives in the validation study reached a maximum of 93% at 120 h of incubation, showing good agreement with the values from the validation experiment of 93.4%, indicating high accuracy of the CCD procedure. When triticale straw, pine wood, and poplar wood were pretreated and hydrolyzed under optimum conditions obtained from the flax shives experiment, the digestibility reached 98.2, 74.8, and 95.7%, respectively, suggesting that the modest pretreatment process using phosphoric acid is an effective method for perennial plants as well as hard wood.     

Ethanolic fermentation of phosphoric acid hydrolysates from olive tree pruning

The objective of this work was to study the feasibility of using phosphoric acid to hydrolyze the hemicellulosic fraction of olive tree pruning, as a step in the bioconversion process to produce ethanol. Milled olive tree pruning was submitted to hydrolysis at 90 °C by phosphoric acid in a concentration range 0.3–8N for 240 min. The hydrolysates were then fermented by Pachysolen tannophilus. The hydrolysis stage was evaluated by the evolution of glucose and reducing sugars generated and by the conversion of hemicellulose fraction. The main parameters determined in the fermentation were: maximum specific growth rate, specific substrate consumption rate, specific ethanol production rate and ethanol yield. The maximum ethanol yield (0.38 kg/kg, equivalent to 74.5% of the theoretical yield) was obtained when hydrolysing with 0.5N phosphoric acid. Hemicellulose conversion is however incomplete at these operational conditions. Higher acid concentrations lead to higher hydrolysis of hemicellulose, but the ethanol yields resulting from the fermentation are lower.     

Processing of materials derived from sweet sorghum for biobased products

Sweet sorghum (Sorghum bicolor (L.) Moench) is particularly suitable as a feedstock for a variety of bioprocesses, largely because of its high yields of both lignocellulosic biomass and fermentable saccharides. Sweet sorghum is less economically important for refined sugar production than other sugar crops, e.g., sugar beet and sugarcane, but can produce more raw fermentable sugar under marginal conditions than those crops. In this review, the agronomic requirements of sorghum (viz., water, soil, and nutrient requirements), cultural practices, and plant morphology are discussed from a bioprocessing perspective. Historically, sugar extraction from the plant in the form of juice has been of primary interest; these methods, along with modern developments are presented. Recently, the direct yeast fermentation of sorghum juice for ethanol production has been studied. Additionally, the bagasse resulting from the juice extraction has been used for a variety of potential products: forage, silage, combustion energy, synthesis gas, and paper. The bagasse contains high levels of relatively low crystallinity cellulose, along with relatively labile lignin, and so is itself of interest as a fermentation feedstock. Whole sorghum stalk, and its bagasse, have been subjected to studies of a wide array of pretreatment, enzymatic hydrolysis, and fermentation processes. The potential fermentation products of sweet sorghum are wide ranging; those demonstrated include ethanol, acetone, butanol, various lipids, lactic acid, hydrogen, and methane. Several potential native products of the plant, in addition to cellulose for paper production, are also identified: waxes, proteins, and allelopathic compounds, such as sorgoleone.     

Biological pretreatment of sugar cane bagasse for the production of cellulases and xylanases by Penicillium echinulatum

In this study, sugar cane bagasse was pretreated with the white rot fungus Pleurotus sajor-caju PS 2001, and this biomass was subsequently used in the production of cellulases and xylanases by the fungus Penicillium echinulatum. Despite the environmental advantages offered by this type of pretreatment, the enzymatic activity obtained with biologically pretreated sugar cane bagasse (PSCB) was lower than that of the control treatments, which were carried out with untreated sugar cane bagasse (SCB) and cellulose. For medium supplemented with PSCB, the average peak activities obtained were 0.13, 1.0, 0.18, and 0.33 U ml^?1 for FPA, endoglucanase, β-glucosidases, and xylanases, respectively. For the cellulose, control values of 0.52, 1.20, 0.20, and 1.46 U ml^?1, and SCB values of 0.95, 1.60, 0.21, and 1.49 U ml^?1 were obtained, respectively. Although the enzymatic activities of the culture with biologically pretreated sugar cane bagasse were lower than the cultures carried out with untreated sugar cane bagasse, it should be noted that production of enzymes of the cellulase and hemicellulase complex after production of the mushrooms is another way to add value to this agricultural residue.     

Sequential extractions and structural characterization of lignin with ethanol and alkali from bamboo (Neosinocalamus affinis)

A sequential process with the combination of ethanol and alkali aqueous solutions was utilized to extract lignin from bamboo (Neosinocalamus affinis), a potential lignocellulosic material. In this case, the successive treatments of dewaxed bamboo with 70% ethanol at 80 °C, 0.2 and 0.5 M NaOH, 70% ethanol containing 0.6 M NaOH, and 1.0, 2.0, and 3.0 M NaOH at 50 °C, resulted in a total yield of acid-insoluble lignin fractions of 10.06%, corresponding to release of 62.25% original lignin from the cell walls. The lignin fractions obtained were then characterized by GPC, FT-IR, NMR spectroscopy, and sugar analysis. As compared to the alkali lignin fractions, the ethanol-soluble lignin fraction had a relatively higher molecular weight (2670 g/mol) and the content of carbohydrates primarily consisted of glucose 2.01% and xylose 1.90%. This suggested that the carbohydrate chains linked to lignin may increase the hydrodynamic volume of lignin and therefore increase the apparent molecular weight of the ethanol-soluble lignin. HSQC spectra analysis revealed that the alkali lignin fractions consisted mainly of β-O-4′ linkages combined with small amounts of β-β′, β-5′, β-1′ linkages, and p-hydroxycinnamyl alcohol end groups. Furthermore, minor amounts of esterified p-coumaric and ferulic acids were also detected in the lignins isolated.     

Sugarcane bagasse whiskers: Extraction and characterizations

This work evaluates the use of sugarcane bagasse (SCB) as a source of cellulose to obtain whiskers. These fibers were extracted after SCB underwent alkaline peroxide pre-treatment followed by acid hydrolysis at 45 °C. The influence of extraction time (30 and 75 min) on the properties of the nanofibers was investigated. Sugarcane bagasse whiskers (SCBW) were analyzed by transmission electron microscopy (TEM), X-ray diffraction (XRD) and thermogravimetric analysis (TGA) in air atmosphere. The results showed that SCB could be used as source to obtain cellulose whiskers and they had needle-like structures with an average length (L) of 255 ± 55 nm and diameter (D) of 4 ± 2 nm, giving an aspect ratio (L/D) around 64. More drastic hydrolysis conditions (75 min) resulted in less thermally stable whiskers and caused some damage on the crystal structure of the cellulose as observed by XRD analysis.     

Optimization of ethanol production from microwave alkali pretreated rice straw using statistical experimental designs by Saccharomyces cerevisiae

Ethanol production from agro-waste provides an alternative energy-production system. Statistical experimental designs were used for optimization of critical nutrients and process variables for ethanol production. The critical nutrients and process variables were initially selected according to a Placket-Burman (PB) design. Selected factors (inoculum level 1-5%, pH 4.5-7, temperature 25-35 °C and urea concentration 0.25-0.75 g/L) were optimized by response surface methodology (RSM) based on a three-level four-factor Box-Behnken design (BBD). Under optimum conditions of inoculum level 3%, pH 5.75, temperature 30 °C and urea concentration 0.50 g/L maximum ethanol production obtained 13.2 g/L from microwave alkali pretreated rice straw with ethanol productivity 0.33 g/L/h. Under optimum conditions ethanol production studied at fermenter level and obtained ethanol concentration 19.2 g/L, ethanol productivity 0.53 g/L/h and ethanol yield to consumed sugar 0.50 g/g. These results indicated that ethanol production can be enhanced by optimization of nutritional and process variables.     

Susceptibility of wheat gluten to enzymatic hydrolysis following deamidation with acetic acid and sensory characteristics of the resultant hydrolysates

The effect of acetic acid and hydrochloric acid (HCl) deamidation pretreatment on the susceptibility of wheat gluten to enzymatic hydrolysis by Pancreatin and sensory characteristics of the resultant hydrolysates was investigated. At two degrees of deamidation (24% and 60%, with or without moisture-heating, respectively), wheat gluten pretreated by acetic acid deamidation was more susceptible to be hydrolyzed as evaluated by the hydrolysis degree, nitrogen solubility index, titratable acid amount and free carbohydrate content of the hydrolysates. Wheat gluten pretreated by acetic acid deamidation at a degree of 24% exhibited the highest susceptibility to enzymatic hydrolysis. Moisture-heating (121 °C, 10 min) in the deamidation pretreatment decreased the susceptibility of wheat gluten to enzymatic hydrolysis and the peptide factions of ≤3000 Da in the hydrolysates due to the formation of larger molecule weight aggregates. The hydrolysates prepared from acetic acid deamidated wheat gluten showed more intense glutamate-like and sauce-scented taste and better nutritional characteristics.     

Optimization of switchgrass and extruder parameters for enzymatic hydrolysis using response surface methodology

Switchgrass is an important biomass that can be hydrolyzed to yield fermentable sugars through pretreatment, which is the primary and expensive step in conversion of biomass to bio-ethanol. Most of the pretreatment operates in batch mode, which is energy intensive, requires high capital, results in decomposition of hemicellulose, and formation of inhibitors. Considering these shortcomings, a novel biomass pretreatment method using a high shear bioreactor could be a viable continuous one. The current study was undertaken to determine the effect of biomass parameters such as moisture content (10, 20, 30, 40, and 50% wb) and particle size (2, 4, 6, 8, and 10 mm) over a range of barrel temperature and screw speed (45-225 °C and 20-200 rpm). Statistical analyses revealed that among the independent variables considered temperature, screw speed, and moisture content had significant effect on sugar recoveries. Proposed quadratic model to predict glucose, xylose, and combined sugar recoveries from switchgrass had a high F and R² values indicating that the model has the ability to represent the relationship among the independent variables studied. The optimum pretreatment condition of barrel temperature 176 °C, screw speed 155 rpm, moisture content 20% wb, and particle size 8 mm resulted in maximum glucose, xylose, and combined sugar recoveries of 41.4, 62.2 and 47.4%, respectively. The optimum pretreated switchgrass had 50% higher surface area than that of the control.