Quang A. Nguyen

Microbial pretreatment of biomass: potential for reducing severity of thermochemical biomass pretreatment.

Typical pretreatment requires high-energy (steam and electricity) and corrosion-resistant, high-pressure reactors. A review of the literature suggests that fungal pretreatment could potentially lower the severity requirements of acid, temperature and time. These reductions in severity are also expected to result in less biomass degradation and consequently lower inhibitor concentrations compared to conventional thermochemical pretreatment. Furthermore, potential advantages of fungal pretreatment of agricultural residues, such as corn stover, are suggested by its effectiveness in improving the cellulose digestibility of many types of forage fiber and agricultural wastes. Our preliminary tests show a three- to five-fold improvement in enzymatic cellulose digestibility of corn stover after pretreatment with Cyathus stercoreus; and a ten- to 100-fold reduction in shear force needed to obtain the same shear rate of 3.2 to 7 rev/s, respectively, after pretreatment with Phanerochaete chrysosporium.

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.

Effects of Temperature and Moisture on Dilute-Acid Steam Explosion Pretreatment of Corn Stover and Cellulase Enzyme Digestibility

Corn stover is emerging as a viable feedstock for producing bioethanol from renewable resources. Dilute-acid pretreatment of corn stover can solubilize a significant portion of the hemicellulosic component and enhance the enzymatic digestibility of the remaining cellulose for fermentation into ethanol. In this study, dilute H2SO4 pretreatment of corn stover was performed in a steam explosion reactor at 160°C, 180°C, and 190°C, approx 1 wt% H2SO4, and 70-s to 840-s residence times. The combined severity (Log10 [Ro] - pH), an expression relating pH, temperature, and residence time of pretreatment, ranged from 1.8 to 2.4. Soluble xylose yields varied from 63 to 77% of theoretical from pretreatments of corn stover at 160 and 180°C. However, yields >90% of theoretical were found with dilute-acid pretreatments at 190°C. A narrower range of higher combined severities was required for pretreatment to obtain high soluble xylose yields when the moisture content of the acid-impregnated feedstock was increased from 55 to 63 wt%. Simultaneous saccharification and fermentation (SSF) of washed solids from corn stover pretreated at 190°C, using an enzyme loading of 15 filter paper units (FPU)/g of cellulose, gave ethanol yields in excess of 85%. Similar SSF ethanol yields were found using washed solid residues from 160 and 180°C pretreatments at similar combined severities but required a higher enzyme loading of approx 25 FPU/g of cellulose.

Dilute acid pretreatment of softwoods

Selective thinning of forests in the western United States will generate a large, sustainable quantity of softwood residues that can be an attractive feedstock for fuel ethanol production. The major species available from thinning of forests in northern California and the eastern Rocky Mountains include white fir (Abies concolor), Douglas fir (Pseudotsuga menziesii), and Ponderosa pine (Pinus ponderosa). Douglas fir chips were soaked in 0.4% sulfuric acid solution, then pretreated with steam at 200 – 230°C for 1 – 5 min. After pretreatment, 90 – 95% of the hemicellulose and as much as 20% of the cellulose was solubilized in water, and 90% of the remaining cellulose can be hydrolyzed to glucose by cellulase enzyme. The prehydrolysates, at as high as 10% total solid concentration, can be readily fermented by the unadapted yeastSaccharomyces cerevisiae D5A.

Dilute acid hydrolysis of softwoods

Whole tree chips obtained from softwood forest thinnings were converted to ethanol via a two-stage dilute acid hydrolysis followed by yeast fermentation. The chips were first impregnated with dilute sulfuric acid, then pretreated in a steam explosion reactor to hydrolyze, more than 90% of the hemicellulose and approx 10% of the cellulose. The hydrolysate was filtered and washed with water to recover the sugars. The washed fibers were then subjected to a second acid im pregnation and hydrolysis to hydrolyze as much as 45% of the reamining cellulose. The liquors from both hydrolysates were combined and fermented to ethanol by a Saccharomyces cerevisiae yeast that had been adapted to the inhibitors. Based on available hexose sugars, ethanol yields varied from 74 to 89% of theoretical. Oligosaccharide contents higher than about 10% of the total available sugar appear to have a negative impact on ethanol yield.

Microbial Pretreatment of Biomass

Typical pretreatment requires high-energy (steam and electricity) and corrosion-resistant, high-pressure reactors. A review of the literature suggests that fungal pretreatment could potentially lower the severity requirements of acid, temperature and time. These reductions in severity are also expected to result in less biomass degradation and consequently lower inhibitor concentrations compared to conventional thermochemical pretreatment. Furthermore, potential advantages of fungal pretreatment of agricultural residues, such as corn stover, are suggested by its effectiveness in improving the cellulose digestibility of many types of forage fiber and agricultural wastes. Our preliminary tests show a three- to five-fold improvement in enzymatic cellulose digestibility of corn stover after pretreatment with Cyathus stercoreus; and a ten- to 100-fold reduction in shear force needed to obtain the same shear rate of 3.2 to 7 rev/s, respectively, after pretreatment with Phanerochaete chrysosporium.

Conversion of distiller's grain into fuel alcohol and a higher-value animal feed by dilute-acid pretreatment.

Over the past three decades ethanol production in the United States has increased more than 10-fold, to approx 2.9 billion gal/yr (mid-2003), with ethanol production expected to reach 5 billion gal/yr by 2005. The simultaneous coproduction of 7 million t/yr of distillers grain (DG) may potentially drive down the price of DG as a cattle feed supplement. The sale of residual DG for animal feed is an important part of corn dry-grind ethanol production economics; therefore, dry-grind ethanol producers are seeking ways to improve the quality of DG to increase market penetration and help stabilize prices. One possible improvement is to increase the protein content of DG by converting the residual starch and fiber into ethanol. We have developed methods for steam explosion, SO2, and dilute-sulfuric acid pretreatment of DG for evaluation as a feedstock for ethanol production. The highest soluble sugar yields (∼77% of available carbohydrate) were obtained by pretreatment of DG at 140°C for 20 min with 3.27 wt% H2SO4. Fermentation protocols for pretreated DG were developed at the bench scale and scaled to a working volume of 809 L for production of hydrolyzed distillers grain (HDG) for feeding trials. The pretreated DG was fermented with Saccharomyces cerevisiae D5A, with ethanol yields of 73% of theoretical from available glucans. The HDG was air-dried and used for turkey-feeding trials. The inclusion of HDG into turkey poult (as a model non-ruminant animal) diets at 5 and 10% levels, replacing corn and soybean meal, showed weight gains in the birds similar to controls, whereas 15 and 20% inclusion levels showed slight decreases (−6%) in weight gain. At the conclusion of the trial, no negative effects on internal organs or morphology, and no mortality among the poults, was found. The high protein levels (58–61%) available in HDG show promising economics for incorporation of this process into corn dry-grind ethanol plants.

Effects of Pressing Lignocellulosic Biomass on Sugar Yield in Two‐Stage Dilute‐Acid Hydrolysis Process

Dilute sulfuric acid catalyzed hydrolysis of biomass such as wood chips often involves pressing the wood particles in a dewatering step (e.g., after acid impregnation) or in compression screw feeders commonly used in continuous hydrolysis reactors. This study addresses the effects of pressing biomass feedstocks using a hydraulic press on soluble sugar yield obtained from two‐stage dilute‐acid hydrolysis of softwood. The pressed acid‐impregnated feedstock gave significantly lower soluble sugar yields than the never‐pressed (i.e., partially air‐dried or filtered) feedstock. Pressing acid‐impregnated feedstocks before pretreatment resulted in a soluble hemicellulosic sugar yield of 76.9% from first‐stage hydrolysis and a soluble glucose yield of 33.7% from second‐stage hydrolysis. The dilute‐acid hydrolysis of partially air‐dried feedstocks having total solids and acid concentrations similar to those of pressed feedstocks gave yields of 87.0% hemicellulosic sugar and 46.9% glucose in the first and second stages, respectively. Microscopic examination of wood structures showed that pressing acid‐impregnated wood chips from 34 to 54% total solids (TS) did not cause the wood structure to collapse. However, pressing first‐stage pretreated wood chips (i.e., feedstock for second‐stage hydrolysis) from approximately 30 to 43% TS caused the porous wood matrix to almost completely collapse. It is hypothesized that pressing alters the wood structure and distribution of acid within the cell cavities, leading to uneven heat and mass transfer during pretreatment using direct steam injection. Consequently, lower hydrolysis yield of soluble sugars results. Dewatering of corn stover by pressing did not impact negatively on the sugar yield from single‐stage dilute‐acid pretreatment.

Dilute acid hydrolysis of softwoods: scientific note.

Whole tree chips obtained from softwood forest thinnings were converted to ethanol via a two-stage dilute acid hydrolysis followed by yeast fermentation. The chips were first impregnated with dilute sulfuric acid, then pretreated in a steam explosion reactor to hydrolyze, more than 90% of the hemicellulose and approx 10% of the cellulose. The hydrolysate was filtered and washed with water to recover the sugars. The washed fibers were then subjected to a second acid im pregnation and hydrolysis to hydrolyze as much as 45% of the reamining cellulose. The liquors from both hydrolysates were combined and fermented to ethanol by a Saccharomyces cerevisiae yeast that had been adapted to the inhibitors. Based on available hexose sugars, ethanol yields varied from 74 to 89% of theoretical. Oligosaccharide contents higher than about 10% of the total available sugar appear to have a negative impact on ethanol yield.

Softwood forest thinnings as a biomass source for ethanol production: a feasibility study for California.

A plan has been put forth to strategically thin northern California forests to reduce fire danger and improve forest health. The resulting biomass residue, instead of being open burned, can be converted into ethanol that can be used as a fuel oxygenate or an octane enhancer. Economic potential for a biomass‐to‐ethanol facility using this softwood biomass was evaluated for two cases: stand‐alone and co‐located. The co‐located case refers to a specific site with an existing biomass power facility near Martell, California. A two‐stage dilute acid hydrolysis process is used for the production of ethanol from softwoods, and the residual lignin is used to generate steam and electricity. For a plant processing 800 dry tonnes per day of feedstock, the co‐located case is an economically attractive concept. Total estimated capital investment is approximately