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Summary of Expansions and Updates in R&D GREET® 2025

The research and development (R&D) version of Greenhouse gases, Regulated Emissions, and Energy use in Technologies (GREET®) model, or R&D GREET, is developed by Argonne National Laboratory (Argonne) with the support of the U.S. Department of Energy (DOE) and other federal agencies. R&D GREET is a life cycle analysis (LCA) model, structured to systematically examine the energy and environmental effects of a wide variety of transportation fuels and vehicle technologies in all transportation sub-sectors (i.e., road, air, marine, and rail) and other end-use sectors, and energy technology systems. Argonne has expanded and updated the model in many areas in R&D GREET 2025. This report provides a summary of the expansions and updates. R&D GREET is designed as an attributional LCA model, but specific consequential effects for certain pathways are included by default when they are scientifically important and/or highly policy relevant. For example, two major consequential elements – induced land use change (ILUC) for biofuels and avoided emissions for certain RNG-based pathways – are implemented by default in R&D GREET because they are highly relevant to our user base and crucial for a comprehensive evaluation of the environmental effects of these technologies. In a future version of R&D GREET, these consequential elements for biofuels and RNG could be more clearly distinguished from core attributional results. We will consider adding a highly visible toggle that would allow users to turn off or adjust these consequential impacts.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Discovery of genes that positively affect biomass and stress associated traits in poplar

Woody biomass serves as a renewable resource for various industries, including pulp and paper production, construction, biofuels, and electricity generation. However, the molecular mechanisms behind biomass traits are poorly understood, which significantly curtails the speed and efficiency of their improvement. We used activation tagging to discover genes that can positively affect tree biomass-associated traits. We generated and screened under greenhouse conditions a population of 2,700 independent activation tagging lines. A total of 761 lines, which had significantly and positively affected at least one biomass-associated trait, were discovered. The tag was positioned in the genome for forty lines which were affected in multiple traits and activation of proximal genes validated for a subset. For two lines we fully recapitulated the phenotype of the original lines through overexpression. Moreover, the overexpression led to more pronounced and additional improvements, not observed in the original lines. Importantly, the overexpression of a Fasciclin-like gene (PtaFLA10) and a Patatin-like gene (PtaPAT) was found to substantially improve biomass, with a 40% increase in dry-stem weight, and enhance drought tolerance, respectively. Additionally, PtaPAT overexpression increased cellulose content, which is crucial for biofuel production. Our work shows that the activation tagging approach applied even on a non-genome saturation scale in a poplar tree can be successfully used for the discovery of genes positively modify biomass productivity. Such dominant forward genetics approaches can aid in biotechnological manipulation of woody biomass traits and help unravel the functions and mechanisms of individual genes, gene families, and regulatory modules.

Plant Sciences↗

The Role of Lignin Molecular Weight on Activated Carbon Pore Structure

Over the past decade, the production of biofuels from lignocellulosic biomass has steadily increased to offset the use of fuels from petroleum. To make biofuels cost-competitive, however, it is necessary to add value to the “ligno-” components (up to 30% by mass) of the biomass. The properties of lignin, in terms of molecular weight (MW), chemical functionality, and mineral impurities often vary from biomass source and biorefinery process, resulting in a challenging precursor for product development. Activated carbon (AC) is a feasible target for the lignin-rich byproduct streams because it can be made from nearly any biomass, and it has a market capacity large enough to use much of the lignin generated from the biorefineries. However, it is not known how the variability in the lignin affects the key properties of AC, because, until now, they could not be well controlled. In this work, various fractions of ultraclean (<0.6% ash) lignin are created with refined MW distributions using Aqueous Lignin Purification using Hot Agents (ALPHA) and used as precursors for AC. AC is synthesized via zinc chloride activation and characterized for pore structure and adsorption capacity. We show that AC surface area and the adsorption capacity increase when using lignin with increasing MW, and, furthermore, that reducing the mineral content of lignin can significantly enhance the AC properties. The surface area of the AC from the highest MW lignin can reach ~1830 m2/g (absorption capacity). Furthermore, single step activation carbonization using zinc chloride allows for minimal carbon burn off (<30%), capturing most of the lignin carbon compared to traditional burn off methods in biorefineries for heat generation.

Biochemistry & Molecular Biology↗

Effects of Media Nutrient Variation on Microalgae Productivity and Economics During Semi-Continuous Cultivation

The development of large-scale microalgae growth for biofuel production is currently limited by the cost of biomass production. However, new approaches to infrastructure and cultivation practices are bringing the field closer to realization. Macronutrients in the cultivation media contribute significant costs, especially since their concentrations have not been optimized for specific strains and conditions. Environmental photobioreactors (ePBRs) were used to simulate cultivation under outdoor conditions, during which the nitrogen and phosphorus levels in the media were varied. The growth of two potential biofuel production strains, Picochlorum celeri and Tetraselmis striata, with varying nutrient inputs during summer and winter scripts, respectively, was studied. This study demonstrated that nitrogen and phosphorus in f/2 media could be reduced by more than 60% from the standard formulation, while maintaining growth rates in a semi-continuous harvesting approach. Experiments comparing the standard and reduced nutrient input concentrations were also conducted for both species in 820 L outdoor raceway ponds, in Mesa, AZ. P. celeri grown in these ponds in October had a growth rate of 10.6 ± 0.7 g/m 2 /day and 10.6 ± 0.3 g/m 2 /day for the standard and low-nutrient P. celeri ponds, respectively. T. striata grown in April–May had a growth rate of 16.6 ± 1.4 g/m 2 /day for the standard nutrient input ponds and 17.4 ± 1.1 g/m 2 /day for the low-nutrient input ponds, and in October 14.5 ± 0.6 g/m 2 /day for standard nutrient ponds and 14.4 ± 0.6 g/m 2 /day for low-nutrient ponds. These outdoor data therefore confirmed the indoor ePBR data. Techno-economic analysis shows that, if high growth rates can be attained at lower nutrient concentrations, a reduction of at least 60% in nutrient costs can be achieved. Such results highlight the importance of managing macronutrient media inputs, as these have a considerable contribution to biomass production costs in large-scale facilities. The analysis also points to the importance of maintaining high spent medium recycling rates in an industrial deployment, so as to minimize the losses of nitrogen and phosphorus compounds.

09 BIOMASS FUELS↗

Analysis of carbon capture at cellulosic biorefineries

The large-scale production of cellulosic biofuels would involve spatially distributed systems including biomass fields, logistics networks and biorefineries. Better understanding of the interactions between landscape-related decisions and the design of biorefineries with carbon capture and storage (CCS) in a supply chain context is needed to enable efficient systems. Here we analyse the cost and greenhouse gas mitigation potential for cellulosic biofuel supply chains in the US Midwest using realistic spatially explicit land availability and crop productivity data and consider fuel conversion technologies with detailed CCS design for their associated CO2 streams.

carbon capture and storage (CCS)↗

Characteristics and Influence of Biosmoke on the Fine-Particle Ionic Composition Measured in Asian Outflow during the Transport and Chemical Evolution Over the Pacific (TRACE-P) Experiment

We investigate the sources, prevalence, and fine-particle inorganic composition of biosmoke over the western Pacific Ocean between 24 February and 10 April 2001. The analysis is based on highly time-resolved airborne measurements of gaseous and fine- particle inorganic chemical composition made during the NASA Transport and Chemical Evolution over the Pacific (TRACE-P) experiment. At latitudes below approximately 25 deg. N, relatively pure biomass burning plumes of enhanced fine-particle potassium, nitrate, ammonium, light-absorbing aerosols, and CO concentrations were observed in plumes that back trajectories and satellite fire map data suggest originated from biomass burning in southeast Asia. Fine-particle water-soluble potassium (K+) is confirmed to be a unique biosmoke tracer, and its prevalence throughout the experiment indicates that approximately 20% of the TRACE-P Asian outflow plumes were influenced, to some extent, by biomass or biofuel burning emissions. At latitudes above 25 deg. N, highly mixed urban/industrial and biosmoke plumes, indicated by SO(sup 2, sub 4) and K+, were observed in 5 out of 53 plumes. Most plumes were found in the Yellow Sea and generally were associated with much higher fine-particle loadings than plumes lacking a biosmoke influence. The air mass back trajectories of these mixed plumes generally pass through the latitude range of between 34 deg. and 40 deg. N on the eastern China coast, a region that includes the large urban centers of Beijing and Tianjin. A lack of biomass burning emissions based on fire maps and high correlations between K+ and pollution tracers (e.g., S(sup 2, sub 4) suggest biofuel sources. Ratios of fine-particle potassium to sulfate are used to provide an estimate of relative contributions of biosmoke emissions to the mixed Asian plumes. The ratio is highly correlated with fine-particle volume (r(sup 2) = 0.85) and predicts that for the most polluted plume encounter in TRACE-P, approximately 60% of the plume is associated with biosmoke emissions. On average, biosmoke contributes approximately 35-40% to the measured fine inorganic aerosol mass in the mixed TRACE-P plumes intercepted north of 25% latitude.

Ma, Y.↗

Aviation Fueling: A Cleaner, Greener Approach

Projected growth of aviation depends on fueling where specific needs must be met. Safety is paramount, and along with political, social, environmental and legacy transport systems requirements, alternate aviation fueling becomes an opportunity of enormous proportions. Biofuels sourced from halophytes, algae, cyanobacteria, and weeds using wastelands, waste water, and seawater have the capacity to be drop-in fuel replacements for petroleum fuels. Biojet fuels from such sources solves the aviation CO2 emissions issue and do not compete with food or freshwater needs. They are not detrimental to the social or environmental fabric and use the existing fuels infrastructure. Cost and sustainable supply remains the major impediments to alternate fuels. Halophytes are the near-term solution to biomass/biofuels capacity at reasonable costs; they simply involve more farming, at usual farming costs. Biofuels represent a win-win approach, proffering as they do at least the ones we are studying massive capacity, climate neutral-to-some sequestration, and ultimately, reasonable costs.

Hendricks, Robert C.↗

Radiative Forcing of the Direct Aerosol Effect from AeroCom Phase II Simulations

We report on the AeroCom Phase II direct aerosol effect (DAE) experiment where 16 detailed global aerosol models have been used to simulate the changes in the aerosol distribution over the industrial era. All 16 models have estimated the radiative forcing (RF) of the anthropogenic DAE, and have taken into account anthropogenic sulphate, black carbon (BC) and organic aerosols (OA) from fossil fuel, biofuel, and biomass burning emissions. In addition several models have simulated the DAE of anthropogenic nitrate and anthropogenic influenced secondary organic aerosols (SOA). The model simulated all-sky RF of the DAE from total anthropogenic aerosols has a range from −0.58 to −0.02 W m(sup−2), with a mean of −0.27 W m(sup−2 for the 16 models. Several models did not include nitrate or SOA and modifying the estimate by accounting for this with information slightly strengthens the mean. Modifying the model estimates for missing aerosol components and for the time period 1750 to 2010 results in a mean RF for the DAE of −0.35 W m(sup−2). Compared to AeroCom Phase I (Schulz et al., 2006) we find very similar spreads in both total DAE and aerosol component RF. However, the RF of the total DAE is stronger negative and RF from BC from fossil fuel and biofuel emissions are stronger positive in the present study than in the previous AeroCom study.We find a tendency for models having a strong (positive) BC RF to also have strong (negative) sulphate or OA RF. This relationship leads to smaller uncertainty in the total RF of the DAE compared to the RF of the sum of the individual aerosol components. The spread in results for the individual aerosol components is substantial, and can be divided into diversities in burden, mass extinction coefficient (MEC), and normalized RF with respect to AOD. We find that these three factors give similar contributions to the spread in results

Computer simulation↗

Radiative Forcing Due to Major Aerosol Emitting Sectors in China and India

Understanding the radiative forcing caused by anthropogenic aerosol sources is essential for making effective emission control decisions to mitigate climate change. We examined the net direct plus indirect radiative forcing caused by carbonaceous aerosol and sulfur emissions in key sectors of China and India using the GISS-E2 chemistry-climate model. Diesel trucks and buses (67 mW/ sq. m) and residential biofuel combustion (52 mW/ sq. m) in India have the largest global mean, annual average forcings due mainly to the direct and indirect effects of BC. Emissions from these two sectors in China have near-zero net global forcings. Coal-fired power plants in both countries exert a negative forcing of about -30 mW/ sq. m from production of sulfate. Aerosol forcings are largest locally, with direct forcings due to residential biofuel combustion of 580 mW/ sq. m over India and 416 mW/ sq. m over China, but they extend as far as North America, Europe, and the Arctic

radiative forcing↗

The Effect of Growth Environment and Salinity on Lipid Production and Composition of Salicornia virginica

Finding a viable and sustainable source of renewable energy is a global task. Biofuels as a renewable energy source can potentially be a viable option for sustaining long-term energy needs. Biodiesel from halophytes shows great promise due to their ability to serve not only as a fuel source, but a food source as well. Halophytes are one of the few biomass plant species that can tolerate a wide range of saline conditions. We investigate the feasibility of using the halophyte, Salicornia virginica as a biofuel source by conducting a series of experiments utilizing various growth and salinity conditions. The goal is to determine if the saline content of Salicornia virginica in our indoor growth vs outdoor growth conditions has an influence on lipid recovery and total biomass composition. We focused on using standard lipid extraction protocols and characterization methods to evaluate twelve Salicornia virginica samples under six saline values ranging from freshwater to seawater and two growth conditions. The overall goal is to develop an optimal lipid extraction protocol for Salicornia virginica and potentially apply this protocol to halophytes in general.

Bomani, Bilal Mark McDowell↗

Bounding the Role of Black Carbon in the Climate System: a Scientific Assessment

Black carbon aerosol plays a unique and important role in Earth's climate system. Black carbon is a type of carbonaceous material with a unique combination of physical properties. This assessment provides an evaluation of black-carbon climate forcing that is comprehensive in its inclusion of all known and relevant processes and that is quantitative in providing best estimates and uncertainties of the main forcing terms: direct solar absorption; influence on liquid, mixed phase, and ice clouds; and deposition on snow and ice. These effects are calculated with climate models, but when possible, they are evaluated with both microphysical measurements and field observations. Predominant sources are combustion related, namely, fossil fuels for transportation, solid fuels for industrial and residential uses, and open burning of biomass. Total global emissions of black carbon using bottom-up inventory methods are 7500 Gg/yr in the year 2000 with an uncertainty range of 2000 to 29000. However, global atmospheric absorption attributable to black carbon is too low in many models and should be increased by a factor of almost 3. After this scaling, the best estimate for the industrial-era (1750 to 2005) direct radiative forcing of atmospheric black carbon is +0.71 W/sq m with 90% uncertainty bounds of (+0.08, +1.27)W/sq m. Total direct forcing by all black carbon sources, without subtracting the preindustrial background, is estimated as +0.88 (+0.17, +1.48) W/sq m. Direct radiative forcing alone does not capture important rapid adjustment mechanisms. A framework is described and used for quantifying climate forcings, including rapid adjustments. The best estimate of industrial-era climate forcing of black carbon through all forcing mechanisms, including clouds and cryosphere forcing, is +1.1 W/sq m with 90% uncertainty bounds of +0.17 to +2.1 W/sq m. Thus, there is a very high probability that black carbon emissions, independent of co-emitted species, have a positive forcing and warm the climate. We estimate that black carbon, with a total climate forcing of +1.1 W/sq m, is the second most important human emission in terms of its climate forcing in the present-day atmosphere; only carbon dioxide is estimated to have a greater forcing. Sources that emit black carbon also emit other short-lived species that may either cool or warm climate. Climate forcings from co-emitted species are estimated and used in the framework described herein. When the principal effects of short-lived co-emissions, including cooling agents such as sulfur dioxide, are included in net forcing, energy-related sources (fossil fuel and biofuel) have an industrial-era climate forcing of +0.22 (0.50 to +1.08) W/sq m during the first year after emission. For a few of these sources, such as diesel engines and possibly residential biofuels, warming is strong enough that eliminating all short-lived emissions from these sources would reduce net climate forcing (i.e., produce cooling). When open burning emissions, which emit high levels of organic matter, are included in the total, the best estimate of net industrial-era climate forcing by all short-lived species from black-carbon-rich sources becomes slightly negative (0.06 W/sq m with 90% uncertainty bounds of 1.45 to +1.29 W/sq m). The uncertainties in net climate forcing from black-carbon-rich sources are substantial, largely due to lack of knowledge about cloud interactions with both black carbon and co-emitted organic carbon. In prioritizing potential black-carbon mitigation actions, non-science factors, such as technical feasibility, costs, policy design, and implementation feasibility play important roles. The major sources of black carbon are presently in different stages with regard to the feasibility for near-term mitigation. This assessment, by evaluating the large number and complexity of the associated physical and radiative processes in black-carbon climate forcing, sets a baseline from which to improve future climate forcing estimates.

black carbon↗

Quantifying Food Security and Mitigation Risks Consequential to Climate Change Impacts on Crop Yields

Climate change is expected to impact crop yields globally, with some regions benefiting from favorable conditions and CO2 fertilization, while others face adverse effects from altered precipitation and higher temperatures. Changes in crop yields can destabilize the global food system and pose challenges to food security. Moreover, crop production is crucial, as biofuels are becoming increasingly important contributors to climate change mitigation measures aimed at limiting global warming. This study uses the Integrated Model to Assess the Global Environment integrated assessment model framework to analyze different indicators related to food security and climate change mitigation under varying climate change impacts on crop yields. Twelve spatially explicit crop productivity projections were taken from the full archive of the Global Gridded Crop Model Intercomparison of 120 climate-crop model combinations, forced by CMIP6-based climate scenarios. The selection includes two average-performing climate-crop model combinations, two pessimistic combinations that perform one standard deviation below the mean, and two optimistic model combinations that perform one standard deviation above the mean. To single out the effect of climate change on productivity changes, we drew samples from two representative concentration pathways (RCP2.6 and RCP8.5). These productivity projections were applied within an otherwise uniform scenario (SSP2) and analyzed for their effect on total calorie demand, crop prices, and number of people at risk of undernourishment to quantify food security. Risks to climate change mitigation targets were explored by modeling the total bioenergy supply, emissions, and global mean temperature. The results revealed significant differences in the risk of food security and mitigation potential between different regions and climate change scenarios. Across scenarios, the crop area extent can vary up to 2 million km2 due to changing crop yields. The projected change in global hunger ranges from 60 to 160 million undernourished people, indicating uncertainty between climate and crop model combinations. Low-income regions are especially impacted because of their high sensitivity to changes in food prices. Global climate change mitigation ambitions can also deviate by the latter part of the 21st century, as changes in yields will impact biofuel production as well as agriculture, forestry and other land use emissions. The quantitative insights generated by this study highlight the need for global policy efforts to make the agricultural system more adaptive to climate change to handle potential negative impacts.

crop yields↗

Discrete-Element and Material-Point Method (DEM and MPM) Based Solvers for Sustainable Technologies

We present the use of discrete element method (DEM) and material point method (MPM) in three relevant green technology applications that include biomass feedstock handling, lithium-ion battery manufacturing, and high-pressure reverse osmosis. Our open-source DEM and MPM solvers are developed using performance portable grid and particle management library, AMReX, thus enabling superior performance on NVIDIA and AMD GPUs with > 100 million particles. Our DEM solver resolves the motion of individual particles in a granular system and includes a bonded sphere method for modeling non-spherical particles along with Hertzian and liquid bridge-based contact models. We simulate highly variable biomass feedstock flows in large-scale hoppers for biofuel production and electrode calendering in battery manufacturing using DEM. Our simulations predict flow blockage in large scale biomass hoppers and electrode microstructure variations, thus providing valuable information for biofuel and battery manufacturers, respectively. The second half of the talk will be on MPM and its application towards pore resolved simulations of reverse osmosis membranes under compressive loads. We present a validation study of our MPM simulations with membrane microscopy imaging thus providing useful insights on membrane stability under high pressure conditions. We also present a spectral stability analysis of using linear hat, quadratic and cubic spline basis in MPM indicating regions of numerical stability.

BIOMASS FUELS,MATHEMATICS AND COMPUTING↗

Herbaceous Feedstock 2022 State of Technology Report

The U.S. Department of Energy promotes production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the state of technology (SOT). As part of its involvement in this mission, Idaho National Laboratory completes an annual SOT report for nth-plant and 1st-plant herbaceous biomass feedstock logistics. The purpose of the SOT is to provide the status of feedstock supply system technology development for herbaceous biomass to biofuels relative to technical targets and cost goals from specific design cases, based on data and experimental results. Although conventional feedstock supply systems form the backbone of the emerging biofuels industry, they have limitations that restrict widespread implementation on a national scale. To meet the demands of the future industry, the feedstock supply system must shift from the conventional system to what has been termed “advanced” supply systems. In advanced designs, a distributed network of aggregation and processing centers, termed “depots,” are employed near the points of biomass production (i.e., the field or forest) to reduce feedstock variability and produce feedstocks of a uniform format, moving toward biomass commoditization. The 2022 Herbaceous SOT is part of a vision of achieving an implemented advanced feedstock supply system, which produces a stable, tradable commodity at the decentralized distributed depot. It utilizes feedstock fractionation by incorporating technologies that can separate the biomass into its anatomical fractions (leaves, husks, stems and cobs) to reduce impurities and produce fractions that satisfy downstream quality considerations. By using a series of air classification steps, this strategy can reduce the extrinsic ash in corn stover and produce enriched tissue fractions that can be blended to a conversion specification or converted individually in optimized biochemical conversion campaigns. Additionally, a majority of the leaves (which do not meet the quality specification) are separated out early and can be supplied to alternate markets. The 2022 Herbaceous SOT incorporates an advanced biomass fractionation and processing system to produce pellets enriched tissues from three-pass corn stover. The resulting enriched pellets are delivered to the biorefinery individually where they can be blended to a specification or converted in campaigns where the conditions are optimized for each tissue. Unused fractions can be sent to a a midstream market or to a different conversion process that is better suited to their properties to offset the cost of the delivered feedstock. The main benefits from the proposed system can be summarized as: (1) $6.86/dry ton (2016$) lower cost for the air classification due to elimination of the requirement to discard the high ash lights fraction; (2) $1.56/dry ton lower delivered cost by selling the unsuitable leaf fraction into the feed market as a midstream co-product (assuming a selling price that is 11% higher than their cost of production); (3) 0.98% increase in carbohydrate content (from 60.16% to 61.14%); and (4) 0.97% decrease in ash content (from 6.00% to 5.03%) compared to the 2021 Herbaceous SOT. Overall, the 2022 nth-plant Herbaceous SOT predicts a modeled delivered feedstock cost of $78.64/dry ton (2016$) if it is assumed that the enriched leaf fraction is sold at its production cost; this is a slight increase of $0.43/dry ton increase from the 2021 Herbaceous SOT nth-Supply case cost. The increased cost derived from a $0.38/dry ton increase in transportation and handling cost to procure more biomass (to replace the enriched leaf fraction that was not delivered to the biorefinery. The total preprocessing cost was $0.27/dry ton higher than the 2021 result because of updates to energy consumption, purchasing price and dry matter loss data for the rotary shear ($3.00/dry ton increase) and the pelleting mill ($4.52/dry ton increase). The data utilized were generated in pilot-scale tests in the Biomass Feedstock National User Facility (BFNUF) at INL and at Forest Concepts, including tests for rotary shear and pelleting of the air classified fractions. A greenhouse gas emissions analysis was performed by Argonne National Laboratory using the most up to date version of the Greenhouse Gases, Regulated Emissions, and Energy use in Transportation model (GREET®). The analysis showed an increase of 17.34 kg CO2e/dry ton from the 2021 SOT (67.71 kg CO2e/ton in the 2021 Herbaceous SOT to 85.05 kg CO2e/ton in the 2022 Herbaceous SOT). The net increase is primarily attributed to increased energy consumption in pelleting mill.

09 BIOMASS FUELS↗

Pyrophosphate-free glycolysis in Clostridium thermocellum increases both thermodynamic driving force and ethanol titers

Background: Clostridium thermocellum is a promising candidate for production of cellulosic biofuels, however, its final product titer is too low for commercial application, and this may be due to thermodynamic limitations in glycolysis. Previous studies in this organism have revealed a metabolic bottleneck at the phosphofructokinase (PFK) reaction in glycolysis. In the wild-type organism, this reaction uses pyrophosphate (PP i ) as an energy cofactor, which is thermodynamically less favorable compared to reactions that use ATP as a cofactor. Previously we showed that replacing the PP i -linked PFK reaction with an ATP-linked reaction increased the thermodynamic driving force of glycolysis, but only had a local effect on intracellular metabolite concentrations, and did not affect final ethanol titer. Results: In this study, we substituted PP i -pfk with ATP-pfk, deleted the other PPi-requiring glycolytic gene pyruvate:phosphate dikinase (ppdk), and expressed a soluble pyrophosphatase (PPase) and pyruvate kinase (pyk) genes to engineer PP i -free glycolysis in C. thermocellum. We demonstrated a decrease in the reversibility of the PFK reaction, higher levels of lower glycolysis metabolites, and an increase in ethanol titer by an average of 38% (from 15.1 to 21.0 g/L) by using PP i -free glycolysis. Conclusions: By engineering PP i -free glycolysis in C. thermocellum, we achieved an increase in ethanol production. These results demonstrate that optimizing the thermodynamic landscape through metabolic engineering can enhance product titers. While further increases in ethanol titers are necessary for commercial application, this work represents a significant step toward engineering glycolysis in C. thermocellum to increase ethanol titers.

09 BIOMASS FUELS↗

Hydrocyclone pre-processing of wastewater algae: A strategy for inorganic ash separation

Microalgae cultivation on wastewater can provide remediation and generate valuable feedstocks for biofuel production. Wastewater algae typically have a high percentage of inorganic ash, which can reduce yield and quality of biocrude produced during hydrothermal liquefaction (HTL). Here, in this work, we evaluated the ability of hydrocyclone pre-processing to remove inorganic ash from wastewater algae. The pH of the algae slurry was adjusted to 9.5 to encourage the formation of precipitates and create a density differential between ash particles and algal cells. Hydrocyclone processing successfully concentrated ash particles in the underflow fraction and reduced the total ash percentage in the overflow fraction. Overall, hydrocyclone processing reduced the total ash by 21%, while only 8% of organics were lost. Elemental and mineral analysis showed that Mg and P were concentrated in the underflow in the form of baricite (an isomorph of vivianite). Future research should focus on improving vivianite and/or baricite formation, and therefore ash removal, by providing a reducing environment. The addition of multiple hydrocyclones in series could also improve the removal of ash. We concluded that hydrocyclone treatment of wastewater algae is a feasible method to remove inorganic ash, but further process optimization is required.

09 - BIOMASS FUELS↗

Adsorptive denitrogenation of model fuel with silica gel

The results establish the adsorption capacity, kinetics, interaction strength of pyridine/indole with sorbents and regeneration, highlighting silica gel's potential as an efficient and cost-effective adsorbent for biofuel nitrogen compounds removal.

09 BIOMASS FUELS↗

Harnessing plant‐based platform for low‐cost cellulosic sugar recovery from bioenergy crops

The price of pure cellulase enzyme for the recovery of fermentable cellulosic sugars is one of the major challenges that limit the commercialization of second-generation biofuels and bio-based products. This work shows a means to greatly reduce the cost of cellulases. The abundant capacity of plants to synthesize and hyperaccumulate transgenic proteins in their plastids has been demonstrated for two gene constructs using either NPT II or Tet C to regulate the expression of bacterial cellulase Cel6A. Previously, the expression of Cel6A to 20–35% of total soluble leaf protein in tobacco was shown in two consecutive field trials. Moving forward, the key challenge was the extraction of active enzymes from the transgenic tobacco leaves hyperaccumulating bacterial cellulases in a cost-effective manner. The study showed that unpurified crude extracts of NPT II and Tet C transgenic tobacco leaves recovered ~23% and ~29% w/w cellulosic sugars from energycane bagasse, respectively. A supplementation of as low as 25% of purified commercial cellulase improved the glucose recovery by 2.9 times (~85% w/w) compared to 100% crude extract, which is comparable to the glucose recovery obtained by commercial cellulases, thereby suggesting a reduced requirement of commercial cellulases leading to a cost reduction of 75% for biorefineries. Assuming a stable 40% cellulase yield in total soluble protein under field conditions and multiple harvests (one to three) a year, the study estimates that the potential cost for saccharification of 1 t of lignocellulosic biomass can be reduced to 67–200 USD by using crude leaf extracts of transgenic tobacco.

biofuel↗