Nicholas J. Nagle

Efficacy of a Hot Washing Process for Pretreated Yellow Poplar to Enhance Bioethanol Production

Cost reductions for pretreatment and bioconversion processes are key objectives necessary to the successful deployment of a bioethanol industry. These unit operations have long been recognized for their impact on the production cost of ethanol. One strategy to achieve this objective is to improve the pretreatment process to produce a pretreated substrate resulting in reduced bioconversion time, lower cellulase enzyme usage, and/or higher ethanol yields. Previous research produced a highly digestible pretreated yellow poplar substrate using a multistage, continuously flowing, very dilute sulfuric acid (0.07% (w/v)) pretreatment. This process reduced the time required for the bioconversion of pretreated yellow poplar sawdust to ethanol. This resulted in a substantially improved yield of ethanol from cellulose. However, the liquid volume requirements, steam demand, and complexity of the flow‐through reactor configuration were determined to be serious barriers to commercialization of that process. A reconfigured process to achieve similar performance has been developed using a single‐stage batch pretreatment followed by a separation of solids and liquids and washing of the solids at a temperatures between 130 and 150 °C. Separation and washing at the elevated temperature is believed to prevent a large fraction of the solubilized lignin and xylan from reprecipitating and/or reassociating with the pretreated solids. This washing of the solids at elevated temperature resulted in both higher recovered yields of soluble xylose sugars and a more digestible pretreated substrate for enzymatic hydrolysis. Key operating variables and process performance indicators included acid concentration, temperature, wash volume, wash temperature, soluble xylose recovery, and performance of the washed, pretreated solids in bioconversion via simultaneous saccharification and fermentation (SSF). Initial results indicated over a 50% increase in ethanol yield at 72 h for the hot washed material as compared to the control (no washing, no separation) and a 43% reduction of in the bioconversion time required for a high ethanol yield from cellulose

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.

High Xylose Yields from Dilute Acid Pretreatment of Corn Stover Under Process-Relevant Conditions

Pretreatment experiments were carried out to demonstrate high xylose yields at high solids loadings in two different batch pretreatment reactors under process-relevant conditions. Corn stover was pretreated with dilute sulfuric acid using a 4-l Steam Digester and a 4-l stirred ZipperClave® reactor. Solids were loaded at 45% dry matter (wt/wt) after sulfuric acid catalyst impregnation using nominal particle sizes of either 6 or 18xa0mm. Pretreatment was carried out at temperatures between 180 and 200xa0°C at residence times of either 90 or 105xa0s. Results demonstrate an ability to achieve high xylose yields (>80%) over a range of pretreatment conditions, with performance showing little dependence on particle size or pretreatment reactor type. The high xylose yields are attributed to effective catalyst impregnation and rapid rates of heat transfer during pretreatment.

Characterization of pilot-scale dilute acid pretreatment performance using deacetylated corn stover

BackgroundDilute acid pretreatment is a promising process technology for the deconstruction of low-lignin lignocellulosic biomass, capable of producing high yields of hemicellulosic sugars and enhancing enzymatic yields of glucose as part of a biomass-to-biofuels process. However, while it has been extensively studied, most work has historically been conducted at relatively high acid concentrations of 1 - 4% (weight/weight). Reducing the effective acid loading in pretreatment has the potential to reduce chemical costs both for pretreatment and subsequent neutralization. Additionally, if acid loadings are sufficiently low, capital requirements associated with reactor construction may be significantly reduced due to the relaxation of requirements for exotic alloys. Despite these benefits, past efforts have had difficulty obtaining high process yields at low acid loadings without supplementation of additional unit operations, such as mechanical refining.ResultsRecently, we optimized the dilute acid pretreatment of deacetylated corn stover at low acid loadings in a 1-ton per day horizontal pretreatment reactor. This effort included more than 25 pilot-scale pretreatment experiments executed at reactor temperatures ranging from 150 – 170°C, residence times of 10 – 20xa0minutes and hydrolyzer sulfuric acid concentrations between 0.15 – 0.30% (weight/weight). In addition to characterizing the process yields achieved across the reaction space, the optimization identified a pretreatment reaction condition that achieved total xylose yields from pretreatment of 73.5%u2009±u20091.5% with greater than 97% xylan component balance closure across a series of five runs at the same condition. Feedstock reactivity at this reaction condition after bench-scale high solids enzymatic hydrolysis was 77%, prior to the inclusion of any additional conversion that may occur during subsequent fermentation.ConclusionsThis study effectively characterized a range of pretreatment reaction conditions using deacetylated corn stover at low acid loadings and identified an optimum reaction condition was selected and used in a series of integrated pilot scale cellulosic ethanol production campaigns. Additionally, several issues exist to be considered in future pretreatment experiments in continuous reactor systems, including the formation of char within the reactor, as well as practical issues with feeding herbaceous feedstock into pressurized systems.

Measurement of the inhibitory potential and detoxification of biomass pretreatment hydrolysate for ethanol production

The Microtox assay represents a rapid, accurate, and reproducible method for determining general microbial toxicity. This assay was used to evaluate the relative toxicity of a variety of hydrolysate samples derived from dilute-acid and alkaline biomass pretreatment. Toxicity is elicited from biomass degradation products, such as furfural, hydroxymethyl furfural, and acetic acid, generated during pretreatment. Microtox results indicate that the pretreatment samples examined ranged from 9 to 71 toxicity units (TU). Correlations of TU and sample absorbance at several wavelengths were evaluated for all sample series. Sample TU values best agreed with absorbance at 230 nm, but the unsatisfactory fit suggests that absorbance should not be used as an absolute measure of sample toxicity.Microtox data for pretreatment hydrolysate samples were correlated with the inhibition experienced by the ethanologenic yeast,Saccharomyces cerevisiae strain D5A, during the simultaneous saccharification and fermentation (SSF) process of pretreated biomass. None of the alkaline pretreatment conditions produced inhibition during SSF (data not shown). However, the acid pretreatment conditions did produce a wide range of inhibitory and noninhibitory hydrolysates. In general, fermentation was inhibited for acid-pretreated hydrolysate samples with values exceeding 45 TU. Preliminary studies that focused on reducing hydrolysate sample toxicity (detoxification) indicate that adding perlite and zeolite had little effect. However, the use of charcoal, a universal flocculent, or ion-exchange resins significantly reduced sample toxicity, holding promise for the efficient bioconversion of pretreated biomass to ethanol. Moreover, the developed toxicity measurement assay can quickly monitor the quality of the pretreatment process. In this way, biomass conversion operation processes can be reliably controlled at the pilot and commercial scales.

Enzymatic Conversion of Xylan Residues from Dilute Acid-Pretreated Corn Stover

Enzymatic conversion of oligomeric xylose and insoluble xylan remaining after effective pretreatment offers significant potential to improve xylan-to-xylose yields while minimizing yields of degredation products and fermentation inhibitors. In this work, a commercial enzyme cocktail is demonstrated to convert up to 70xa0% of xylo-oligomers found in dilute acid-pretreated hydrolyzate liquor at varying levels of dilution when supplemented with accessory enzymes targeting common side chains. Commercial enzyme cocktails are also shown to convert roughly 80xa0% of insoluble xylan remaining after effective high-solids, dilute acid pretreatment.

Pretreatment technology for the beneficial biological reuse of municipal sewage sludges

Modern municipal sewage waste treatment plants use conventional mechanical and biological processes to reclaim waste waters. This process has the overall effect of converting a water pollution problem into a solid waste disposal problem (sludges). The costs for conventional disposal of sewage sludges have risen dramatically because of increased environmental mandates, which restrict their disposal, as well as a dwindling number of landfills. Previously, we determined that secondary bioprocessing (specifically anaerobic digestion) was not effective in reducing the organic content and bulk of the sludge waste (1). Therefore, we have examined the potential of a variety of pretreatment technologies designed to disrupt the macrostructure of the sludge and thereby enhance its subsequent biodegradation. Two thermal/mechanical pretreatments tested were found to have a dramatic effect on the subsequent bioconversion of the microbial sludges. Both technologies evaluated, sonication and shear, were found to be affected by sludge solids levels, duration of treatment, and treatment temperature. Optimum sonication pretreatment occurred with sludge solids of 1% and treatment times of 4–8 min. The most effectivee treatment temperature tested was 55°C. The optimum enhancement in bioconversion potential for the sonication pretreatment was 80–83% of the materials carbon oxygen demand (COD) content. The optimum shear pretreatment occurred with sludge solids of 1–2% and treatment times of 6–10 min. The most effective treatment temperature tested was 87°C. The optimum enhancement in bioconversion potential for the shear pretreatment was 88–90% of the material’s COD content. These data were the basis for US patent no. 5,380,445, granted January 10, 1995.

A laboratory-scale pretreatment and hydrolysis assay for determination of reactivity in cellulosic biomass feedstocks

BackgroundThe rapid determination of the release of structural sugars from biomass feedstocks is an important enabling technology for the development of cellulosic biofuels. An assay that is used to determine sugar release for large numbers of samples must be robust, rapid, and easy to perform, and must use modest amounts of the samples to be tested.In this work we present a laboratory-scale combined pretreatment and saccharification assay that can be used as a biomass feedstock screening tool. The assay uses a commercially available automated solvent extraction system for pretreatment followed by a small-scale enzymatic hydrolysis step. The assay allows multiple samples to be screened simultaneously, and uses only ~3xa0g of biomass per sample. If the composition of the biomass sample is known, the results of the assay can be expressed as reactivity (fraction of structural carbohydrate present in the biomass sample released as monomeric sugars).ResultsWe first present pretreatment and enzymatic hydrolysis experiments on a set of representative biomass feedstock samples (corn stover, poplar, sorghum, switchgrass) in order to put the assay in context, and then show the results of the assay applied to approximately 150 different feedstock samples covering 5 different materials. From the compositional analysis data we identify a positive correlation between lignin and structural carbohydrates, and from the reactivity data we identify a negative correlation between both carbohydrate and lignin content and total reactivity. The negative correlation between lignin content and total reactivity suggests that lignin may interfere with sugar release, or that more mature samples (with higher structural sugars) may have more recalcitrant lignin.ConclusionsThe assay presented in this work provides a robust and straightforward method to measure the sugar release after pretreatment and saccharification that can be used as a biomass feedstock screening tool. We demonstrated the utility of the assay by identifying correlations between feedstock composition and reactivity in a population of 150 samples.

Demonstration-scale evaluation of a novel high-solids anaerobic digestion process for converting organic wastes to fuel gas and compost

Early evaluations of the bioconversion potential for combined wastes such as tuna sludge and sorted municipal solid waste (MSW) were conducted at laboratory scale and compared conventional low-solids, stirred-tank anaerobic systems with the novel, high-solids anaerobic digester (HSAD) design. Enhanced feedstock conversion rates and yields were determined for the HSAD system. In addition, the HSAD system demonstrated superior resiliency to process failure. Utilizing relatively dry feedstocks, the HSAD system is approximately one-tenth the size of conventional low-solids systems. In addition, the HSAD system is capable of organic loading rates (OLRs) on the order of 20-25 g volatile solids per liter digester volume per d (gVS/L/d), roughly 4-5 times those of conventional systemsCurrent efforts involve developing a demonstration-scale (pilot-scale) HSAD system. A two-ton/d plant has been constructed in Stanton, CA and is currently in the commissioning/startup phase. The purposes of the project are to verify laboratory- and intermediate-scale process performance; test the performance of large-scale prototype mechanical systems; demonstrate the long-term reliability of the process; and generate the process and economic data required for the design, financing, and construction of full-scale commercial systems. This study presents conformational fermentation data obtained at intermediate-scale and a snapshot of the pilot-scale project

The Effect of Biomass Densification on Structural Sugar Release and Yield in Biofuel Feedstock and Feedstock Blends

In this work, we examined the behavior of feedstock blends and the effect of axa0specific feedstock densification strategy (pelleting) on the release and yield of structural carbohydrates in a laboratory-scale dilute acid pretreatment (PT) and enzymatic hydrolysis (EH) assay. We report overall carbohydrate release and yield from the two-stage PT-EH assay for five single feedstocks (two corn stovers, miscanthus, switchgrass, and hybrid poplar) and three feedstock blends (corn stover-switchgrass, corn stover-switchgrass-miscanthus, and corn stover-switchgrass-hybrid poplar). We first examined the experimental results over time to establish the robustness of the PT-EH assay, which limits the precision of the experimental results. The use of two different control samples in the assay enabled us to identify (and correct for) a small bias in the EH portion of the combined assay for some runs. We then examined the effect of variable pretreatment reaction conditions (residence time, acid loading, and reactor temperature) on the conversion of a single feedstock (single-pass corn stover, CS-SP) in order to establish the range of pretreatment reaction conditions likely to provide optimal conversion data. Finally, we applied the assay to the 16 materials (8 feedstocks in 2 formats, loose and pelleted) over a more limited range of pretreatment experimental conditions. The four herbaceous feedstocks behaved similarly, while the hybrid poplar feedstock required higher pretreatment temperatures for optimal results. As expected, the yield data for three blended feedstocks were the average of the yield data for the individual feedstocks. The pelleting process appears to provide a slightly positive effect on overall total sugar yield.