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All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Water Quality Effects of Economically Viable Land Use Change in the Mississippi River Basin under the Renewable Fuel Standard

Demand for biofuel production driven by the Renewable Fuel Standard (RFS2) has coincided with increased land in corn production and increasing nitrogen (N) loss to the Gulf of Mexico. Diversifying cropland with perennial energy crops (miscanthus and switchgrass) may reduce N loss and improve water quality. However, the extent of these benefits depends on the mix of biomass feedstocks (corn stover, perennials) incentivized by the RFS2 and the extent to which energy crops displace N-intensive row crops. We developed an integrated economic-biophysical model to quantify the water quality impacts of three potential policy scenarios that provided corn ethanol at levels before the RFS2 (RFS1 baseline); 15 billion gallons of corn ethanol (corn ethanol only); or 16 billion gallons of cellulosic ethanol in addition to corn ethanol (corn + cellulosic ethanol). Our results showed that economically optimal locations for perennial energy crop production were distributed across idle cropland with lower intrinsic N loss than active cropland. We found stover removal incentivized by the RFS2 offset N loss benefits of perennial energy crops. Furthermore, this finding suggests that targeted incentives for N loss reduction are needed to supplement the RFS2 to induce displacement of N-intensive row crops with energy crops to reduce N losses.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2024

Trade can buffer climate-induced risks and volatilities in crop supply

Climate change is intensifying the frequency and severity of extreme events, posing challenges to food security. Corn, a staple crop for billions, is particularly vulnerable to heat stress, a primary driver of yield variability. While many studies have examined climate impact on average corn yields, little attention has been given to the climate impact on production volatility. This study investigates the future volatility and risks associated with global corn supply under climate change, evaluating the potential benefits of two key adaptation strategies: irrigation and market integration. A statistical model is employed to estimate corn yield response to heat stress and utilize NEX-GDDP-CMIP6 climate data to project future production volatility and risks of substantial yield losses. Three metrics are introduced to quantify these risks: Sigma (σ), the standard deviation of year-on-year yield change, which reflects overall yield volatility; Rho (ρ), the risk of substantial loss, defined as the probability of yield falling below a critical threshold; and Beta (β), a relative risk coefficient that captures the volatility of a region's corn production compared to the globally integrated market. The analysis reveals a concerning trend of increasing year-on-year yield volatility (σ) across most regions and climate models. This volatility increase is significant for key corn-producing regions like Brazil and the United States. While irrigated corn production exhibits a smaller rise in volatility, suggesting irrigation as a potential buffer against climate change impacts, it is not a sustainable option as it can cause groundwater depletion. On the other hand, global market integration reduces overall volatility and market risks significantly with less sustainability concerns. Furthermore, these findings highlight the importance of a multidimensional approach to adaptation in the food sector. While irrigation can benefit individual farmers, promoting global market integration offers a broader solution for fostering resilience and sustainability across the entire food system.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗
DOE OSTI2017

Spatial Analysis of Stover Moisture Content During Harvest Season in the U.S.

The moisture content of a maturing crop varies as the harvest season progresses. For crop residues such as corn stover, moisture content at the time of harvest can be as high as 75% (wet mass basis) to less than 20% depending on the geographic location (climate conditions) and stage of harvest. Biomass moisture content is critical for baling and extended storage. It is therefore essential to have an estimate of the quantities of corn stover available as wet or dry for various parts of the U.S. To this end, we analyzed hourly weather data (temperature, humidity, and rainfall) from the Typical Meteorological Year v.3 (TMY3) dataset developed by the National Renewable Energy Laboratory. A recently published set of equations for calculating the moisture content of stover as a function of hourly temperature, humidity, and rainfall were used. The annual start and end of corn grain harvest along with annual grain production (in bushels) for each state were extracted from USDA National Agricultural Statistics Service reports. Using these datasets and moisture sorption equations, the percentage of corn stover tonnage with moisture content less than 20%, between 20% and 40%, or greater than 40% was estimated from the length of time that the biomass was in these moisture content ranges. These calculations were carried out for several locations within each of the states for which TMY data were available. It was concluded that about 37.2% of corn stover is dry (<20% moisture content), whereas 36.5% is wet (>40% moisture content) nationwide. The remaining 27.0% of corn stover is between 20% and 40% moisture content. Keywords: Corn stover, Equilibrium moisture content, Field drying, Moisture content, Stover harvest, Typical Meteorological Year data.

09 BIOMASS FUELS↗
DOE OSTI2019

Data from: Early accumulation of active fraction soil carbon in newly established cellulosic biofuel systems

We examined relative changes in soil C pools shortly after the establishment of six perennial and two annual bioenergy cropping systems that differed in diversity (monoculture vs. polyculture). Perennial systems included two monocultures (switchgrass, Panicum virgatum; and miscanthus, Miscanthus × giganteus) and four polycultures including hybrid poplar (Populus sp.) + herbaceous understory; mixed native grasses, successional vegetation, and restored prairie. Two annual systems included no-till continuous corn (Zea mays) and rotational corn (corn-soybean (Glycine max)-canola (Brassica napus)). Each crop was planted in a full factorial design at both a moderate fertility Alfisol and a high fertility Mollisol site. Relative differences in active, slow, and passive C pools in surface soils, where C changes are most likely to be detected early, were evaluated with 322-day laboratory incubations followed by acid hydrolysis to infer different pools from exponential decay curves. Five years post-establishment, active C pools under perennial polycultures at the Alfisol site were up to twice those under annual and perennial monocultures, and followed the order hybrid poplars (696 ± 216 μg C g− 1 soil, n = 5 replicate blocks) ≈ native grasses (656 ± 155) ≈ restored prairie (638 ± 44) > early successional (500 ± 54) ≫ continuous corn (237 ± 68) ≈ rotational corn (180 ± n.a.). Active C pools in perennial monocultures were similar to those in continuous corn: switchgrass (274 ± 29) ≈ miscanthus (299 ± 9). In contrast, differences in active C pools among crops at the more fertile Mollisol site were not detectable except for greater pools in the restored prairie and rotational corn systems. At both sites, slow and passive C pools differed little among systems except that slow pools were greater in the poplar system. That diversity rather than perenniality itself led to greater active C pools suggests that polycultures might be used to accelerate soil C accumulation in bioenergy and other perennial cropping systems.

09 BIOMASS FUELS↗
DOE OSTI2019

Supply and value chain analysis of mixed biomass feedstock supply system for lignocellulosic sugar production

Here the sustainable supply of high-quality, low-cost biomass feedstock to cellulosic biorefineries is still a challenge. Utilizing corn stover as the single feedstock to a biorefinery also poses a source of risk to seasonal feedstock availability, affordability, and sustainability. Implementing a year-round feedstock switching strategy or utilizing an optimal mixture of corn stover, miscanthus, and switchgrass is demonstrated to diminish these concerns while reducing the cost of sugar production by as much as 3.8% and 13.6%, respectively, when compared to a baseline sugar price of 441.9 $ metric ton-1 (t) attributed to a fixed corn stover feedstock basis. This study determines that a mixing ratio of corn stover, miscanthus, and switchgrass of 36%, 50%, and 14%, respectively, minimizes the selling price of sugar. If miscanthus is sufficiently available in the future, its fraction could be further increased to 70% by reducing the fraction of corn stover. We find that the mixed feedstock considered in this study reduces the nutrient replacement and feedstock transportation costs and improves the quality of feedstock (determined by total carbohydrates and ash) relative to corn stover alone. Results suggest that a high-quality feedstock with a high carbohydrate content is an important metric for consideration, beyond strictly the feedstock cost, to reduce the selling price of sugar and its uncertainties. In the future this may be supported by the increased availability of higher carbohydrate feedstocks such as miscanthus.

09 BIOMASS FUELS↗
DOE OSTI2025

Field–scale evaluation of ecosystem service benefits of bioenergy switchgrass

Purpose-grown perennial herbaceous species are nonfood crops specifically cultivated for bioenergy production and have the potential to secure bioenergy feedstock resources while enhancing ecosystem services. This study assessed soil greenhouse gas emissions (CO 2 and N 2 O), nitrate (NO 3 -N) leaching reduction potential, evapotranspiration (ET), and water-use efficiency (WUE) of bioenergy switchgrass (Panicum virgatum L.) in comparison to corn (Zea mays L.). The study was conducted on field-scale plots in Urbana, IL, during the 2020–2022 growing seasons. Switchgrass was established in 2020 and urea-fertilized at 56 kg N ha –1 year –1 . Corn management followed best management practices for the US Midwest, including no-till and 202 kg N ha –1 year –1 fertilization, applied as urea–ammonium nitrate (32%). Our results showed lower direct N 2 O emissions in switchgrass compared to corn. Although soil CO 2 emissions did not differ significantly during the establishment year, emissions in subsequent years were over 50% higher in switchgrass than in corn, likely due to increased belowground biomass, which was over five times higher in switchgrass. Nitrate-N leaching decreased as the switchgrass stand matured, reaching 80% lower than in corn by the third year. Differences in ET and WUE between corn and switchgrass were not significant; however, results indicate a trend toward reduced WUE in switchgrass under drought, driven by lower aboveground biomass production. Our study demonstrates that switchgrass can be implemented at a commercial scale without negatively impacting the hydrological cycle, while potentially reducing N losses through nitrate-N leaching and soil N 2 O emissions, and enhancing belowground C storage.

09 BIOMASS FUELS↗
DOE OSTI2020

Simulated Biomass Sorghum GHG Reduction Potential is Similar to Maize

Policy support for cellulosic biofuels is contingent on their achieving much greater reductions in life-cycle greenhouse gas emissions than corn starch ethanol. Biomass sorghum has been suggested as a genetically and agronomically tractable feedstock species to augment near-term cellulosic feedstock production. This study used DayCent modeling to investigate biomass sorghum production emissions relative to corn with and without stover utilization at 3,265 across the rainfed United States. Sorghum produced greater average feedstock dry matter (15.6±1.4 vs. 14.8±2.2 Mg ha -1 yr -1 ) and slightly lower estimated ethanol energy yields (10.6±1.0 vs. 11.8±2.9 MJ m -2 yr -1 ) as corn grain with 75% stover collection. The high biomass removals in both the sorghum and corn stover scenarios led to soil organic carbon losses on 90% and 100% of sites, respectively. Average feedstock production emissions intensities were similar between sorghum and corn with 75% stover removal (17.6±2.8 vs. 18.8±3.0 g CO 2 e MJ -1 ), but were notably lower under sorghum for sites in the southwestern study region (13.6±3.0 vs. 22.5±3.1 g CO 2 e MJ -1 ). Futhermore, these results suggest that biomass sorghum produces cellulosic feedstock with similar emissions to corn grain and at current yield levels is unlikely to meet the Renewable Fuel Standard emissions reduction threshold for cellulosic biofuels.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

Long-term variability of root production in bioenergy crops from ingrowth core measurements

Long-term determination of root biomass production upon land-use conversion to biofuel crops is rare. To assess land-use legacy influences on belowground biomass accumulation, we converted 22-year-old Conservation Reserve Program (CRP) grasslands and 50+-year-old agricultural (AGR) lands to corn (C), switchgrass (Sw) and restored prairie (Pr) biofuel crops. We maintained one CRP grassland as a reference (Ref). We hypothesized that land-use history and crop type have significant effects on root density, with perennial crops on CRP grasslands having a higher root biomass productivity, while corn grown on former agricultural lands produce the lowest root biomass. Methods The ingrowth core method was used to determine in situ ingrowth root biomass, alongside measurements of aboveground net primary productivity (ANPP). Ancillary measurements, including air temperature, growing season length and precipitation were used to examine their influences on root biomass production. Important Findings Root biomass productivity was the highest in unconverted CRP grassland (1716 g m-2 yr-1) and lowest in corn fields (526 g m-2 yr-1). All perennial sites converted from CRP and AGR lands had lower root biomass and ANPP in the first year of planting but peaked in 2011 for switchgrass and a year later for restored prairies. Ecosystem stability was higher in restored prairies (AGR-Pr: 4.3 ± 0.11; CRP-Pr: 4.1 ± 0.10), with all monocultures exhibiting a lower stability. Root biomass production was positively related to ANPP (R2 = 0.40). Overall, attention should be given to root biomass accumulation in large-scale biofuel production as it is a major source of carbon sequestration.

Environmental Sciences & Ecology↗
DOE OSTI2025

Data for "Field-scale Evaluation of Ecosystem Service Benefits of Bioenergy Switchgrass"

Purpose-grown perennial herbaceous species are nonfood crops specifically cultivated for bioenergy production and have the potential to secure bioenergy feedstock resources while enhancing ecosystem services. This study assessed soil greenhouse gas emissions (CO2 and N2O), nitrate (NO3-N) leaching reduction potential, evapotranspiration (ET), and water-use efficiency (WUE) of bioenergy switchgrass ( Panicum virgatum L.) in comparison to corn ( Zea mays L.). The study was conducted on field-scale plots in Urbana, IL, during the 2020–2022 growing seasons. Switchgrass was established in 2020 and urea-fertilized at 56 kg N ha−1 year−1. Corn management followed best management practices for the US Midwest, including no-till and 202 kg N ha−1 year−1 fertilization, applied as urea–ammonium nitrate (32%). Our results showed lower direct N2O emissions in switchgrass compared to corn. Although soil CO2 emissions did not differ significantly during the establishment year, emissions in subsequent years were over 50% higher in switchgrass than in corn, likely due to increased belowground biomass, which was over five times higher in switchgrass. Nitrate-N leaching decreased as the switchgrass stand matured, reaching 80% lower than in corn by the third year. Differences in ET and WUE between corn and switchgrass were not significant; however, results indicate a trend toward reduced WUE in switchgrass under drought, driven by lower aboveground biomass production. Our study demonstrates that switchgrass can be implemented at a commercial scale without negatively impacting the hydrological cycle, while potentially reducing N losses through nitrate-N leaching and soil N2O emissions, and enhancing belowground C storage.

field data↗
DOE OSTI2017

Biomass Production a Stronger Driver of Cellulosic Ethanol Yield than Biomass Quality

Core Ideas Fermentable sugars were greatest in corn stover > perennial grasses > polycultures. Corn stover had the highest ethanol content. Miscanthus had the highest ethanol yield potential on a per hectare basis. Ethanol yield potential per hectare of switchgrass ≥ corn stover. Biomass yield was the strongest driver of per hectare ethanol yield. Many crops have been proposed as feedstocks for the emerging cellulosic ethanol industry, but information is lacking about the relative importance of feedstock production and quality. We compared yield and sugar content for seven bioenergy cropping systems in south‐central Wisconsin (ARL) and southwestern Michigan (KBS) during three growing seasons (2012 through 2014). The cropping systems were (i) continuous corn stover ( Zea mays L.), (ii) switchgrass ( Panicum virgatum L.), (iii) giant miscanthus ( Miscanthus × giganteus Greef & Deuter ex Hodkinson & Renvoize), (iv) hybrid poplar ( Populus nigra × P. maximowiczii A. Henry ‘NM6’), (v) native grass mix, (vi) early successional community, and (vii) restored prairie. A high‐throughput pretreatment and fermentation assay showed corn stover with the highest sugar content (213 g glucose kg −1 [Glc] and 115 g xylose kg −1 [Xyl]) followed by the two monoculture perennial grass treatments (154 [Glc] and 88 [Xyl]) and then the herbaceous polycultures (135 [Glc] and 77 [Xyl]). Biomass production and sugar content were combined to calculate ethanol yields. Miscanthus had the highest per hectare ethanol yields (1957 l ha −1 yr −1 ARL, 2485 l ha −1 yr −1 KBS) followed by switchgrass (1091 l ha −1 yr −1 ARL, 1017 l ha −1 yr −1 KBS) and corn stover (1121 l ha −1 yr −1 ARL, 878 l ha −1 yr −1 KBS). Perennial grass cropping systems (i.e., switchgrass and miscanthus) had higher per hectare ethanol yields at both sites relative to diverse systems that included dicots. Despite feedstock differences in fermentable sugars, biomass production was the strongest driver of per hectare ethanol yield.

09 BIOMASS FUELS↗
DOE OSTI2019

Stover Harvest did not Change Nitrous Oxide Emissions in Two Minnesota Fields

Corn (Zea mays L.) is grown across vast acreages producing massive quantities of stover making corn a desirable cellulosic bioenergy feedstock. Nitrous oxide (N 2 O) is a potent greenhouse gas (GHG) so small changes in direct soil N 2 O emissions may have substantial influence on global warming potential (GWP) from the agricultural sector. Harvesting stover alters soil properties such as soil moisture, oxygen availability, temperature, and substrate availability (C and N). Thus, harvesting stover might reduce soil N 2 O emissions by reducing substrate and by warming and drying the soil. The goal of the study was to determine how harvesting corn stover altered soil N 2 O emissions. Therefore, soil N 2 O emissions were measured for four crop-years (planting to planting) in two independent studies, one in a field managed without tillage (NT1995) and the other was in a field tilled annually with a chisel plow (Chisel). In this study, each field was in a corn–soybean (Glycine max L. [Merr.]) rotation, with (i) corn grain (Grain) only, (ii) grain plus about 50% of the stover harvested (Grain+Moderate), and (iii) grain plus harvesting as much stover as possible (Grain+Aggressive) treatments. Cumulative soil N 2 O emissions did not differ among treatments in either field during any of the crop-years monitored. Flux events occurred corresponding to fertilizer applications and to spring freeze-thaw events. Cumulative fertilizer stimulated emissions tended to be greater for corn than soybean because of N-fertilizer application. These results are valuable to modelers for enhancing estimates of the N 2 O component of the stover management C-footprint.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2019

Biomass Grinding Process Optimization Using Response Surface Methodology and a Hybrid Genetic Algorithm

Biomass could be a key source of renewable energy. Agricultural waste products, such as corn stover, provide a convenient means to replace fossil fuels, such as coal, and a large amount of feedstock is currently available for energy consumption in the U.S. This study has two main objectives: (1) to understand the impact of corn stover moisture content and grinder speed on grind physical properties; and (2) develop response surface models and optimize these models using a hybrid genetic algorithm. The response surface models developed were used to draw surface plots to understand the interaction effects of the corn stover grind moisture content and grinder speed on the grind physical properties and specific energy consumption. The surface plots indicated that a higher corn stover grind moisture content and grinder speed had a positive effect on the bulk and tapped density. The final grind moisture content was highly influenced by the initial moisture content of the corn stover grind. Optimization of the response surface models using the hybrid genetic algorithm indicated that moisture content in the range of 17 to 19% (w.b.) and a grinder speed of 47 to 49 Hz maximized the bulk and tapped density and minimized the geomantic mean particle length. The specific energy consumption was minimized when the grinder speed was about 20 Hz and the corn stover grind moisture content was about 10% (w.b.).

09 BIOMASS FUELS↗
DOE OSTI2019

Data from: Ecosystem carbon exchange on conversion of Conservation Reserve Program grasslands to annual and perennial cropping systems

Land use changes into and out of agricultural production may substantially influence ecosystem carbon (C) balance for many years. We examined ecosystem C balances for eight years after the conversion of 22 year-old Conservation Reserve Program (CRP) grasslands and formerly tilled agricultural fields (AGR) to annual (continuous no-till corn) and perennial (switchgrass and restored prairie) cropland. An unconverted CRP field (CRP-Ref) was maintained as a historical reference. Ecosystem C balance was assessed using adjusted net ecosystem carbon exchange (NEEadj) calculated by adding C removed in harvested biomass to NEE measured using eddy covariance method. The cumulative NEEadj of the corn and perennial systems on former CRP fields showed that these systems were a net C source to the atmosphere over the 8-year period while on former AGR fields, the perennial systems were net C sinks and the corn system near-neutral. The CRP-Ref was near neutral until a drought year when it became a net source. The corn system on the CRP field will likely reach a new lower soil C equilibrium at least 14 years after conversion but will never regain the C lost upon conversion under current no-till management with residue partially removed. On the other hand, the perennial systems could fully regain in ~14 years the C lost following conversion. The cumulative NEEadj of the corn systems exhibited a higher C emission than did the perennial systems within the same land use histories, reflecting the dominant role of crop type and management in agricultural ecosystem C balance. Results suggest that converting croplands to grasslands results in immediate C gains whereas converting grasslands to croplands results in permanent (no-till corn with partial residue removal) or temporary (perennial herbaceous crops) net C loss to the atmosphere. This has a significant implications for global climate change mitigation where biomass production from annual and perennial crops is promoted to avoid fossil-fuel C emissions (biofuel) or to remove CO2 from the atmosphere (bioenergy C capture and storage).

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS1983

Remote sensing of agricultural crops and soils

Research in the correlative and noncorrelative approaches to image registration and the spectral estimation of corn canopy phytomass and water content is reported. Scene radiation research results discussed include: corn and soybean LANDSAT MSS classification performance as a function of scene characteristics; estimating crop development stages from MSS data; the interception of photosynthetically active radiation in corn and soybean canopies; costs of measuring leaf area index of corn; LANDSAT spectral inputs to crop models including the use of the greenness index to assess crop stress and the evaluation of MSS data for estimating corn and soybean development stages; field research experiment design data acquisition and preprocessing; and Sun-view angles studies of corn and soybean canopies in support of vegetation canopy reflection modeling.

Bauer, M. E.↗
DOE OSTI2021

Is satellite Sun-Induced Chlorophyll Fluorescence more indicative than vegetation indices under drought condition?

Droughts represent one of the most severe abiotic stress factors that could result in great crop yield loss. Numerous vegetation indices have been proposed for monitoring the vegetation condition under stress and assessing drought impacts on yield loss. However, the understanding and comparison between traditional vegetation indices (VIs) and the newly emerging satellite Sun-Induced Chlorophyll Fluorescence (SIF) for monitoring vegetation condition is still limited especially under drought stress and at multiple spatial scales. In this study, the potential of satellite observation SIF for monitoring corn response to drought was investigated based on the 2012 drought in the US Corn Belt. The standardized precipitation evapotranspiration index (SPEI) was used here to quantify drought. We found that all SPEI were above –1, except for July (–1.27), August (–1.39) and September (–1.14) in 2012, indicating the severity of this drought. We examined the relationship between satellite measurements of SIF, SIF yield , VIs (e.g., NDVI and EVI) and SPEI. Results indicated that SIF yield was sensitive to drought and SIF captured the stress more accurately both at the regional and state scales for the US Corn Belt. Quantitatively, SIF yield had a high correlation with SPEI (r = 0.987, p < 0.05) over the entire Corn Belt, and it indicated losses in response to drought approximately one month earlier than SIF/NDVI/EVI. Furthermore, our results demonstrated that SIF could be trusted as an effective indicator to study the relationship between GPP (R 2 ≥ 0.8664, p < 0.01) under drought conditions across the Corn Belt. Finally, this study highlighted the advantage of using satellite SIF observations to monitor the drought stress on crop growth especially GPP at regional scale.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Soil phosphorus drawdown by perennial bioenergy cropping systems in the Midwestern US

Abstract Without fertilization, harvest of perennial bioenergy cropping systems diminishes soil nutrient stocks, yet the time course of nutrient drawdown has not often been investigated. We analyzed phosphorus (P) inputs (fertilization and atmospheric deposition) and outputs (harvest and leaching losses) over 7 years in three representative biomass crops—switchgrass ( Panicum virgatum L.), miscanthus ( Miscanthus × giganteus ) and hybrid poplar trees ( Populus nigra × P. maximowiczii )—as well as in no‐till corn (maize; Zea mays L.) for comparison, all planted on former cropland in SW Michigan, USA. Only corn received P fertilizer. Corn (grain and stover), switchgrass, and miscanthus were harvested annually, while poplar was harvested after 6 years. Soil test P (STP; Bray‐1 method) was measured in the upper 25 cm of soil annually. Harvest P removal was calculated from tissue P concentration and harvest yield (or annual woody biomass accrual in poplar). Leaching was estimated as total dissolved P concentration in soil solutions sampled beneath the rooting depth (1.25 m), combined with hydrological modeling. Fertilization and harvest were by far the dominant P budget terms for corn, and harvest P removal dominated the P budgets in switchgrass, miscanthus, and poplar, while atmospheric deposition and leaching losses were comparatively insignificant. Because of significant P removal by harvest, the P balances of switchgrass, miscanthus, and poplar were negative and corresponded with decreasing STP, whereas P fertilization compensated for the harvest P removal in corn, resulting in a positive P balance. Results indicate that perennial crop harvest without P fertilization removed legacy P from soils, and continued harvest will soon draw P down to limiting levels, even in soils once heavily P‐fertilized. Widespread cultivation of bioenergy crops may, therefore, alter P balances in agricultural landscapes, eventually requiring P fertilization, which could be supplied by P recovery from harvested biomass.

09 BIOMASS FUELS↗
DOE OSTI2022

Declining carbohydrate solubilization with increasing solids loading during fermentation of cellulosic feedstocks by Clostridium thermocellum: documentation and diagnostic tests

Abstract Background For economically viable 2nd generation biofuels, processing of high solid lignocellulosic substrate concentrations is a necessity. The cellulolytic thermophilic anaerobe Clostridium thermocellum is one of the most effective biocatalysts for solubilization of carbohydrate harbored in lignocellulose. This study aims to document the solubilization performance of Clostridium thermocellum at increasing solids concentrations for two lignocellulosic feedstocks, corn stover and switchgrass, and explore potential effectors of solubilization performance. Results Monocultures of Clostridium thermocellum demonstrated high levels of carbohydrate solubilization for both unpretreated corn stover and switchgrass. However, fractional carbohydrate solubilization decreases with increasing solid loadings. Fermentation of model insoluble substrate (cellulose) in the presence of high solids lignocellulosic spent broth is temporarily affected but not model soluble substrate (cellobiose) fermentations. Mid-fermentation addition of cells ( C. thermocellum ) or model substrates did not significantly enhance overall corn stover solubilization loaded at 80 g/L, however cultures utilized the model substrates in the presence of high concentrations of corn stover. An increase in corn stover solubilization was observed when water was added, effectively diluting the solids concentration mid-fermentation. Introduction of a hemicellulose-utilizing coculture partner, Thermoanaerobacterium thermosaccharolyticum, increased the fractional carbohydrate solubilization at both high and low solid loadings. Residual solubilized carbohydrates diminished significantly in the presence of T. thermosaccharolyticum compared to monocultures of C. thermocellum, yet a small fraction of solubilized oligosaccharides of both C 5 and C 6 sugars remained unutilized. Conclusion Diminishing fractional carbohydrate solubilization with increasing substrate loading was observed for C. thermocellum -mediated solubilization and fermentation of unpretreated lignocellulose feedstocks. Results of experiments involving spent broth addition do not support a major role for inhibitors present in the liquid phase. Mid-fermentation addition experiments confirm that C. thermocellum and its enzymes remain capable of converting model substrates during the middle of high solids lignocellulose fermentation. An increase in fractional carbohydrate solubilization was made possible by (1) mid-fermentation solid loading dilutions and (2) coculturing C. thermocellum with T. thermosaccharolyticum , which ferments solubilized hemicellulose. Incomplete utilization of solubilized carbohydrates suggests that a small fraction of the carbohydrates is unaffected by the extracellular carbohydrate-active enzymes present in the culture.

09 BIOMASS FUELS↗
DOE OSTI2017

Ethanol from Agriculture for Arkansas and America

The purpose of this project was to develop technology that would facilitate production of sugars from agricultural residues to enable biofuels and biobased product manufacturing. Our primary technology is to use genetic engineering to put bacterial and fungal cellulase genes into corn kernels, using the grain as the production system for the enzymes. At the beginning of this DoE funded program, we were producing two cellulases—E1 endocellulase from a bacterium found in a hot spring at Yellowstone National Park, and CBH I exocellulase from a wood rot fungus. Our team developed several new regulatory sequences (promoters) that increased enzyme protein accumulation in two kernel compartments (embryo and endosperm). We were also able to capitalize on the diverse genetics of corn to increase protein accumulation. High oil germplasm in particular was instrumental in this increase. A second task in the program was to produce enzymes and proteins that enhanced the activity of the E1 and CBH I enzymes. Our team produced CBH II, from the same wood rot fungus at a level that enabled highly enhanced deconstruction activity of E1 and CBH I in a synergistic manner. We analyzed an additional protein, expansin from cucumber that was expressed in the maize grain expression system. This protein had been previously shown to enhance cellulase activity (D. Cosgrove, Penn State University), and required a large-scale production platform. Our team showed that the corn production system allows industrial amounts of active expansin to be harvested from the grain. One of the challenges of any new production system is to maximize recovery of active ingredient from the raw materials at a cost compatible with its final use. Our team showed that low pH extraction of grain solubilized the enzymes without contamination of native corn protein and active product could be concentrated through ultrafiltration. The final outcomes of this project were the following: 3 cellulase enzymes and the synergistic protein expansin produced at high levels in corn grain, new promoters and combinations of promoters to enhance protein accumulation in grain, application of unique germplasm pools to enhance protein accumulation, and highly efficient processing enabling cost-effective production of cellulases that are highly active in biomass deconstruction.

09 BIOMASS FUELS↗