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At least 73 records · Page 4

Increasing the Carbohydrate Output of Bamboo Using a Combinatorial Pretreatment

Bamboo represents a family of highly lignified biomass with significant resistance to cellulolytic enzyme degradation, and typically, a severe thermochemical pretreatment is needed to overcome its recalcitrance for producing fermentable sugars. Combinatorial pretreatments with a low holding temperature and low alkali dosage were investigated in this study in an attempt to remove the impediments to the accessibility of bamboo cellulose to enzymes and enable the efficient conversion of hemicellulose. Upon autohydrolysis at 180 °C, more than half of the hemicellulose (~58%) was solubilized, contributing to further delignification in the subsequent dilute alkali extraction (DAE). The solubilized hemicellulose mainly existed in the form of soluble xylan, which, after a novel endo-β-1,4-xylanase hydrolysis, generated a large amount of xylo-oligosaccharides with the xylobiose and xylotriose proportion reaching 72.99%. After the fractionation with combinatorial pretreatments, cellulose and xylose enzymatic hydrolysis yields were 73.36 and 86.98%, respectively, representing increments of 22.42 and 43.15% compared with the sole autohydrolysis process. Fluorescence microscopy (FM) and confocal Raman microscopy (CRM) images revealed that these dramatic improvements were caused by the deconstruction of the bamboo cell walls, including separation of the adjacent cell walls, removal of hemicellulose and lignin, migration and redistribution of lignin, and the increased exposure of cellulose. In addition, lignin chemical structure variations during the combinatorial pretreatment were analyzed with gel permeation chromatography (GPC), 2D HSQC NMR, and 31 P NMR, and the correlation between the lignin structure and enzymatic hydrolysis efficiency was investigated. In summary, this study highlighted the application of combinatorial pretreatments to maximize the carbohydrate output of bamboo and unveiled their mechanism that enabled the high-efficiency utilization of bamboo.

09 BIOMASS FUELS↗

Recoveries of Oil and Hydrolyzed Sugars from Corn Germ Meal by Hydrothermal Pretreatment: A Model Feedstock for Lipid-Producing Energy Crops

Vegetable oil is extracted from oil rich seeds, such as soybeans. Genetic engineering of green plants to accumulate oil in vegetative tissue is a future source of oil that promises increased land productivity and the use of marginal lands. However, the low concentration of lipids in current engineered plant biomass samples makes the oil extraction process challenging and expensive. In this study, liquid hot water (LHW) pretreatment was investigated to enhance oil recovery from the solids and increase enzymatic hydrolysis efficiency of such feedstocks. Corn germ meal was chosen as a model feedstock representing lipid-producing energy crops. Germ meal was pretreated at 160 and 180 °C for 10 and 15 min at 20% w/w solids loading. Enzymatic hydrolysis on the pretreated solid was performed. After pretreatment, the oil concentration increased by 2.2 to 4.2 fold. The most severe pretreatment condition of LHW, at 180 °C for 15 min, gave the maximum oil concentration (9.7%, w/w), the highest triacylglycerol (TAG) content of the extracted oil (71.6%), and the highest conversions of glucose and xylose (99.0% and 32.8%, respectively). This study demonstrates that the optimal pretreatment condition for corn germ meal is 180 °C LHW for 15 min. Pretreatment improves lipids recovery from oil bearing biomass with little or no effect on the lipid profile.

09 BIOMASS FUELS↗

Effect of H 2 O on the ethylene glycol/alkali dismantling of bagasse for high-value conversion

Using a high-boiling alcohol system to dismantle main components of biomass is a feasible technology. Reducing dismantle operating costs and improving dismantle efficiency are essential for promoting the green, economical, and sustainable development of biomass refining. Therefore, based on the low cost and chemical properties of H 2 O at high temperature, the effects of different H 2 O dosages in NaOH-catalyzed ethylene glycol (HBAA) system on the dismantling efficiency of bagasse, surface lignin coverage, recovered-lignin activity and enzymatic hydrolysis efficiency were investigated. Compared with the HBAA dismantling system without H 2 O, the HBAA system with 60% w/v H 2 O can obviously increase the removal rates of lignin and hemicellulose, while recovering up to 99% of cellulose and significantly declining surface lignin coverage, thus enhancing the enzymatic hydrolysis efficiency. Additionally, the results of density functional theory calculations and 2D HSQC NMR analysis prove that the synergy between H 2 O and ethylene glycol can promote the esterification reaction occurrence at the α-C carbon cation in β-O-4 structure of lignin, thereby protecting the β-O-4 aromatic ether bond. Simultaneously, when the H 2 O dosage increase from 0% to 60%, the enzymatic yield increases from 84.51% to 93.74% with an enzyme load of 10 FPU/g. Based on experimental results, this study conducted a techno-economic analysis of bagasse dismantling for ethanol and co-production of lignin, achieving a minimum ethanol selling price of $\$$1.07 per kg. Here, in this study, a green and economical solution for dismantling the main components of bagasse is developed, which is important for the high-value conversion of bagasse.

Ethylene glycol↗

Impact of non-irrigation on 1G and 2G bioethanol potential of oilcane feedstock: A field to fuel pipeline study

This study evaluates the bioethanol potential in response to irrigation (IR) and non-irrigation (NIR) of oilcane (OC) during a seasonal drought prior harvest. The juice was extracted through mechanical pressing of stems and fermented by Ethanol Red® yeast to produce first-generation bioethanol. Hydrothermal pretreatment followed by enzymatic hydrolysis of bagasse was performed to produce monomeric sugars from structural carbohydrates. The hydrolysates were fermented with engineered yeast for second-generation bioethanol production. The irrigated oilcane juice (276.3 ± 8.9 g/L) constitutes higher sugar concentrations than non-irrigated oilcane juice (236.5 ± 2.2 g/L). The enzymatic hydrolysis of IR-OC and NIR-OC pretreated bagasse yielded similar concentrations of 247.5 ± 2.22 and 249.7 ± 4.98 g/L fermentable sugars. Industry-relevant bioethanol titers of ≥99 g/L and ≥75 g/L were achieved from juice and hydrolysates, respectively. Therefore, the non-irrigation regime did not impact the 1G and 2G bioethanol titers. However, the overall bioethanol yield can be lower due to the reduction of stem yield (8 %) per hectare.

09 BIOMASS FUELS↗

Data for Impact of Non-Irrigation on 1G and 2G Bioethanol Potential of Oilcane Feedstock: A Field to Fuel Pipeline Study

This study evaluates the bioethanol potential in response to irrigation (IR) and non-irrigation (NIR) of oilcane (OC) during a seasonal drought prior harvest. The juice was extracted through mechanical pressing of stems and fermented by Ethanol Red® yeast to produce first-generation bioethanol. Hydrothermal pretreatment followed by enzymatic hydrolysis of bagasse was performed to produce monomeric sugars from structural carbohydrates. The hydrolysates were fermented with engineered yeast for second-generation bioethanol production. The irrigated oilcane juice (276.3 ± 8.9 g/L) constitutes higher sugar concentrations than non-irrigated oilcane juice (236.5 ± 2.2 g/L). The enzymatic hydrolysis of IR-OC and NIR-OC pretreated bagasse yielded similar concentrations of 247.5 ± 2.22 and 249.7 ± 4.98 g/L fermentable sugars. Industry-relevant bioethanol titers of ≥99 g/L and ≥75 g/L were achieved from juice and hydrolysates, respectively. Therefore, the non-irrigation regime did not impact the 1G and 2G bioethanol titers. However, the overall bioethanol yield can be lower due to the reduction of stem yield (8 %) per hectare.

Biomass Analytics↗

Enhanced Feedstock Characterization and Modeling to Facilitate Optimal Preprocessing and Deconstruction of Corn Stover (Final Report)

This project addresses the challenge of processing corn stover by fractionating this biomass feedstock to both streamline processing and generate new potential co-products. Additionally, the project developed new field-deployable analytical tools that can be coupled with empirical models that were used to predict feedstock properties and processing performance. The overall scope of this project was: (1) identify conditions for optimal corn stover fractionation using a two- stage physical fractionation, (2) assess how physical fractionation impacts properties, partitioning of biomass, and response to processing, (3) further adapt, develop, and validate several advanced characterization tools for assessing biomass properties that can be linked to processing behavior, and (4) develop and validate predictive models based on measurements that can be performed “in the field” or “at the biorefinery gate” to predict feedstock processing behavior (preprocessing and deconstruction). The first objective employed pre-separation processing (size reduction) which was next subjected to enhanced separations to yield fractions enriched or depleted in select compositional components or properties. For the second objective, fractions were screened for their response to post-separation processing (pretreatment and enzymatic hydrolysis). Detailed characterization profiles were developed and dynamic image analysis to assess distribution of particle size and morphology. For the final objective, we utilized these tools to develop empirical models to assess the relative abundance of tissue type in order to assess fractionation efficacy and to predict fraction performance during pretreatment and enzymatic hydrolysis.

09 BIOMASS FUELS↗

Production of chitosan-based composite film reinforced with lignin-rich lignocellulose nanofibers from rice husk

Lignocellulosic nanofibers (LCNFs), implying lignin-containing cellulose fibers, maintain the properties of both lignin and cellulose, which are hydrophobic and hydrophilic, respectively. The presence of hydrophobic lignin in LCNFs is expected to be an economical and attractive option that can improve the thermal and mechanical properties of polymers. Thus, this study was conducted to produce lignin-rich LCNFs from sugar-rich waste obtained from rice husks after acidic pretreatment. The LCNFs were produced from the lignin-rich solid fractions obtained after pretreatment and enzymatic hydrolysis, which were then incorporated as an additive into a chitosan-based film. The variations in lignin content in the range of approximately 50.6%-66.8% in differently obtained LCNFs gave significantly different optical strengths and mechanical properties. These controllable processes may allow for customized film formation. Additionally, the glucose-rich liquid fractions obtained after pretreatment and enzymatic hydrolysis were used as a substrate for ethanol fermentation to achieve total utilization of rice husk biomass waste. In conclusion, the lignin-rich biomass fraction holds promise as a suitable material for chitosan-LCNF film and has the potential to increase the economic feasibility of the biomaterial industry.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Sustainable strategies to achieve industrial ethanol titers from different bioenergy feedstocks: scale-up approach for better ethanol yield

Hydrothermal pretreatment is a promising approach to lignocellulosic biomass processing for enzymatic hydrolysis and high-yield bioethanol fermentation, as it reduces downstream inhibitor content and the amount of toxic byproducts generated. In this paper, the ethanol yield and productivity of an engineered xylose-fermenting strain of Saccharomyces cerevisiae were tested on lignocellulosic hydrolysates produced with varying citrate buffer concentration, solid loading, supplemental nitrogen source, and feedstock of origin, and a semi-integrated bioprocess which integrates enzymatic hydrolysis and bioethanol fermentation was developed. The greatest ethanol yields (g p /g s ) of 0.490 ± 0.008, 0.460 ± 0.001, 0.420 ± 0.002 and 0.410 ± 0.002 were obtained from bioenergy sorghum (BES), Miscanthus × giganteus (MG), energy cane (EC), and oilcane (OC), respectively. In addition, an equivalent of 291 L, 253.54 L, 257.8 L, and 260.3 L of bioethanol were produced per ton of BES, MG, EC, and OC, respectively, by using urea as a nitrogen source in a bioreactor.

09 BIOMASS FUELS↗

Coupling AFEX and steam-exploded sugarcane residue pellets with a room temperature CIIII-activation step lowered enzyme dosage requirements for sugar conversion

In this study, the potential integration of steam explosion (StEx) and ammonia fiber expansion (AFEX) with existing sugar/ethanol mills to form decentralized pre-processing depots was explored. Both StEx and AFEX pretreatment facilitated the production of sugarcane bagasse (SCB) and cane leaf matter (CLM) pellets with significantly higher bulk density, mechanical durability, and hydrophobicity relative to their untreated biomass pellet controls. However, ethanol production from standalone StEx and AFEX-treated SCB and CLM pellets required enzyme dosages greater than 21 mg/g glucan to achieve enzymatic hydrolysis sugar yields of 75% and ethanol titres greater than 40 g.L -1 . Coupling AFEX-treated SCB or CLM pellets with a room temperature CIII I -activation step using liquid ammonia lowered enzyme dosage requirements by more than 50% without affecting ethanol titers and production yields (greater than300 L per Mg residual dry matter raw dry biomass (RDM)). In contrast, treating StEx-treated pellets with CIIII-activation using liquid ammonia did not result in similar enzyme dosage reductions, due to pseudo-lignin formation, leading to enzyme deactivation and/or lignin blockage that retarded enzymatic hydrolysis at low enzyme dosages. A gross energy conversion assessment revealed that low enzyme dosage (3-4 mg enzyme/g RDM) ethanol and electricity co-production from AFEX and CIII I -activated SCB and CLM can recover up to 73% of the energy in the untreated biomass, compared to 54% recovered by StEx and CIII I -activation. The results from this work suggest that StEx or AFEX based pre-processing depots can produce dense and mechanically durable biomass pellets. The AFEX-treated pellets can be easily upgraded using a room temperature CIII I -activation step at the biorefinery to significantly reduce bioconversion enzymes.

42 ENGINEERING↗

Investigation of a Lignin-Based Deep Eutectic Solvent Using p -Hydroxybenzoic Acid for Efficient Woody Biomass Conversion

Deep eutectic solvents (DESs) are effective solvents for biomass conversion and have been proposed as alternatives to ionic liquids (ILs). In this paper, we first report an effective pretreatment of woody biomass using a novel hardwood lignin-based DES prepared from p -hydroxybenzoic acid (PB) and choline chloride (ChCl), which are used as the hydrogen bond donor and acceptor, respectively. The impacts of the ChCl-PB DES on structural properties and enzymatic hydrolysis of poplar wood were comprehensively evaluated. The performance of ChCl-PB DES was also compared with two other DESs with p -coumaric acid (PCA) and 4-hydroxybenzaldehyde (PHA) as the hydrogen bond donors, which have been successfully applied to herbaceous biomass conversion in previous studies. The results showed that hemicellulose and lignin were effectively fractionated during the proposed ChCl-PB DES pretreatment under a relatively mild temperature. The ChCl-PB DES pretreatment resulted in up to 69% of delignification, and the pretreated poplar residues had over 90% of glucose yield by a 72 h-enzymatic hydrolysis. Transformation of poplar biomass during the DES pretreatment was evaluated by gel permeation chromatography (GPC), two-dimensional heteronuclear single quantum coherence (2D-HSQC), and 31 P nuclear magnetic resonance (NMR) spectroscopies. The recovered lignin showed relatively high purity, narrow molecular weight distribution (PDI < 1.6), and relatively low molecular weight (~1400 g/mol for M w ). A sustainable process was achieved by recycling DES as well as the utilization of PB, which is a producible aromatic compound from hardwood lignin, for the proposed DES formation in the system. This study indicates that the proposed novel DES with a hardwood lignin-based PB compound is a promising pretreatment solvent to achieve an economically feasible biomass conversion process.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Data for Tolerance of Engineered Rhodosporidium toruloides to Sorghum Hydrolysates During Batch and Fed-Batch Lipid Production

Oleaginous yeasts are a promising candidate for the sustainable conversion of lignocellulosic feedstocks into fuels and chemicals, but their growth on these substrates can be inhibited as a result of upstream pretreatment and enzymatic hydrolysis conditions. Previous studies indicate a high citrate buffer concentration during hydrolysis inhibits downstream cell growth and ethanol fermentation in Saccharomyces cerevisiae . In this study, an engineered Rhodosporidium toruloides strain with enhanced lipid accumulation was grown on sorghum hydrolysate with high and low citrate buffer concentrations. Both hydrolysis conditions resulted in similar sugar recovery rates and concentrations. No significant differences in cell growth, sugar utilization rates, or lipid production rates were observed between the two citrate buffer conditions during batch fermentation of R. toruloides . Under fed-batch growth on low-citrate hydrolysate a lipid titer of 16.7 g/L was obtained. Citrate buffer was not found to inhibit growth or lipid production in this engineered R. toruloides strain, nor did reducing the citrate buffer concentration negatively affect sugar yields in the hydrolysate. As this process is scaled-up, $131 per ton of hydrothermally pretreated biomass can be saved by use of the lower citrate buffer concentration during enzymatic hydrolysis.

Conversion↗

Tolerance of engineered Rhodosporidium toruloides to sorghum hydrolysates during batch and fed-batch lipid production

Abstract Background Oleaginous yeasts are a promising candidate for the sustainable conversion of lignocellulosic feedstocks into fuels and chemicals, but their growth on these substrates can be inhibited as a result of upstream pretreatment and enzymatic hydrolysis conditions. Previous studies indicate a high citrate buffer concentration during hydrolysis inhibits downstream cell growth and ethanol fermentation in Saccharomyces cerevisiae . In this study, an engineered Rhodosporidium toruloides strain with enhanced lipid accumulation was grown on sorghum hydrolysate with high and low citrate buffer concentrations. Results Both hydrolysis conditions resulted in similar sugar recovery rates and concentrations. No significant differences in cell growth, sugar utilization rates, or lipid production rates were observed between the two citrate buffer conditions during batch fermentation of R. toruloides . Under fed-batch growth on low-citrate hydrolysate a lipid titer of 16.7 g/L was obtained. Conclusions Citrate buffer was not found to inhibit growth or lipid production in this engineered R. toruloides strain, nor did reducing the citrate buffer concentration negatively affect sugar yields in the hydrolysate. As this process is scaled-up, $131 per ton of hydrothermally pretreated biomass can be saved by use of the lower citrate buffer concentration during enzymatic hydrolysis. Graphical Abstract

09 BIOMASS FUELS↗

Studying the Suitability of Nineteen Lignins as Partial Polyol Replacement in Rigid Polyurethane/Polyisocyanurate Foam

In this study, nineteen unmodified lignins from various sources (hardwood, softwood, wheat straw, and corn stover) and isolation processes (kraft, soda, organosolv, sulfite, and enzymatic hydrolysis) were used to replace 30 wt.% of petroleum-based polyol in rigid polyurethane/polyisocyanurate (PUR/PIR) foam formulations. Lignin samples were characterized by measuring their ash content, hydroxyl content (Phosphorus Nuclear Magnetic Resonance Spectroscopy), impurities (Inductively Coupled Plasma), and pH. After foam formulation, properties of lignin-based foams were evaluated and compared with a control foam (with no lignin) via cell morphology, closed-cell content, compression strength, apparent density, thermal conductivity, and color analysis. Lignin-based foams passed all measured standard specifications required by ASTM International C1029-15 for type 1 rigid insulation foams, except for three foams. These three foams had poor compressive strengths, significantly larger cell sizes, darker color, lower closed-cell contents, and slower foaming times. The foam made with corn stover enzymatic hydrolysis lignin showed no significant difference from the control foam in terms of compressive strength and outperformed all other lignin-based foams due to its higher aliphatic and p-hydroxyphenyl hydroxyl contents. Lignin-based foams that passed all required performance testing were made with lignins having higher pH, potassium, sodium, calcium, magnesium, and aliphatic/p-hydroxyphenyl hydroxyl group contents than those that failed.

59 BASIC BIOLOGICAL SCIENCES↗

Conversion of High-Solids Hydrothermally Pretreated Bioenergy Sorghum to Lipids and Ethanol Using Yeast Cultures

Glucose and xylose are the major sugars in cellulosic hydrolysates. The cellulosic sugars can be used for the production of biofuels and value-added bioproducts. In this study, lipid and ethanol were produced from bioenergy sorghum syrups using engineered yeasts. Here, bioenergy sorghum was hydrothermally pretreated at 50% solids loading in a continuous reactor system and mechanically refined sequentially using a burr mill to improve biomass accessibility for hydrolysis. Fed-batch enzymatic hydrolysis was conducted with 50% w/v solids loading to achieve 230 g/L sugar concentration. Different strains of Rhodosporidium toruloides were used to ferment sugars into lipids, and the highest lipid yield of 9.2 g/L was observed. The lipid yield was improved to 19.0 g/l by implementing a two-stage culture where once the sugars were exhausted in the first stage, the yeast was introduced into fresh hydrolysate without adding nitrogen. For ethanol production, the engineered Saccharomyces cerevisiae SR8ΔADH6 was utilized to coferment glucose and xylose. Additionally, the effects of nutrient media (YP, YNB/urea, and urea), cellulosic sugar concentration, and sulfite addition were investigated to optimize the ethanol yield from sorghum syrups. The optimal ethanol yield at 73.3% was obtained from the YNB/urea culture consisting of 34 g glucose/L and 17 g xylose/L.

09 BIOMASS FUELS↗

Data for Conversion of High-Solids Hydrothermally Pretreated Bioenergy Sorghum to Lipids and Ethanol Using Yeast Cultures

Glucose and xylose are the major sugars present in cellulosic hydrolysates. The cellulosic sugars can be used for the production of platform chemicals. In this study, productions of lipid and ethanol by yeasts were compared for concentrated bioenergy sorghum syrup. Bioenergy sorghum was hydrothermally pretreated at 50% w/w solids in a continuous industrial reactor and sequentially mechanically refined using a burr mill to improve biomass accessibility for hydrolysis. Fed-batch enzymatic hydrolysis was conducted with 50% w/v solids loading and cellulase cocktail (50 FPU/g biomass) to achieve 230 g/L sugar concentration. Various strains of Rhodosporidium toruloides were evaluated for converting sugars into lipids, and strain Y-6987 had the highest lipid titer (9.2 g/L). The lipid titer was improved to 19.0 g/L by implementing a two-stage culture scheme, where the first stage was optimized for yeast growth and the second for lipid production. For ethanol production, the engineered Saccharomyces cerevisiae SR8ΔADH6 was used to coferment glucose and xylose. Ethanol fermentation was optimized for media nutrients (YP, YNB/urea, and urea), cellulosic sugar concentration, and sulfite conditioning to maximize the ethanol concentration from sorghum syrups. Fermentation of 70% v/v concentrated hydrolysate conditioned with sulfite produces 50.1 g/L ethanol from 141 g/L of sugars.

Conversion↗

Microwave Pretreatment For Hydrolysis Of Cellulose

Microwave pretreatment enhances enzymatic hydrolysis of cellulosic wastes into soluble saccharides used as feedstocks for foods, fuels, and other products. Low consumption of energy, high yield, and low risk of proposed hydrolysis process incorporating microwave pretreatment makes process viable alternative to composting.

Cullingford, Hatice S.↗

Process strategies for recovery of sugars, lipids, and lignin from oilcane bagasse using natural deep eutectic solvents (NADES)

Sugarcane is being enhanced as a bioenergy crop by engineering it to accumulate and store lipids along with polymeric sugars in vegetative tissues. However, there is no existing process that allows for processing this new crop to recover both lipid and cellulosic sugars from the oilcane bagasse. Therefore, a comprehensive investi- gation of two pretreatment methods—natural deep eutectic solvents (NADES) and chemical-free hydrothermal pretreatment (HT) was conducted to judge their suitability for recovering fermentable sugars, lipids, and lignin from bagasse. Two NADES, i.e., choline chloride: lactic acid (ChCl:LA) and betaine: lactic acid (BT:LA) were prepared using a 1:2 M ratio and were evaluated for pretreatment of oilcane bagasse at 10, 20, and 50 % (w/w) solids, followed by enzymatic hydrolysis at 10 % (w/w) solids. Notably, ChCl:LA NADES treatment at 10 % (w/ w) solids at 140 °C for 2 h, solubilized 78.8 % of lignin and 80.4 % of hemicellulose and allowed 82.7 % enzymatic conversion of glucans to glucose. In contrast, HT pretreatment removed approximately 87.6 % of the hemicellulose and provided an enzymatic glucose yield of 69.7 %. Furthermore, ChCl:LA operated at 50 % solids loading the enriched lipids 2.6-fold (9.2 wt%) in recovered solids compared to HT (6.4 %) and BT:LA (5.1 %) pretreatment processes. NMR-HSQC and GPC analysis showed that ChCl:LA also cleaved the most lignin β–O–4 linkages and demonstrated lower molecular weight compared to HT. As a result, this study demonstrates that NADES pretreatment is an effective green processing method for recovering lipids, sugars, and lignin from bioenergy crops at high solid loading (50 % w/w) within the context of an integrated biorefinery.

09 BIOMASS FUELS↗

Feedstock/Pretreatment Screening for Bioconversion of Sugars and Lignin Residues

This project will conduct biomass deconstruction (pretreatment and enzymatic saccharification) on two representative biomass feedstocks and four different pretreatment processes, including three high temperature steam/chemical pretreatments and one low temperature chemical/mechanical pretreatment. A third biomass feedstock will undergo biomass deconstruction with three different pretreatment processes, including two high temperature chemical pretreatments and one low temperature chemical/mechanical pretreatment. Several pretreatment conditions will be performed in a screening study using NLR pilot-scale pretreatment equipment to generate a range of pretreated biomass substrates. A selected number of these substrates will be chosen for enzymatic saccharification evaluation, based on standard compositional analysis of the pretreated substrates as a primary indicator of pretreatment efficacy. Resulting enzymatic hydrolysis slurries will be analyzed to determine overall biomass sugar yields. Additional compositional analysis will be performed to determine oligomeric sugar composition and structure, to analyze structural characteristics of solids fractions on native, pretreated, and enzymatically saccharified biomass residues for one of the biomass feedstocks, corn stover. The enzymatically saccharified materials will undergo 2,3-butanediol fermentation in a shaker flask as bench scale. Using relevant process performance data collected in these various conversion steps, technoeconomic analysis activities will be performed to compare the economic potential of the various biomass feedstock and pretreatment processes and to identify key economic drivers and sustainability metrics.

09 BIOMASS FUELS↗