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DOE OSTI2024

Structural Characterization and Dynamics of AdhE Ultrastructures from Clostridium thermocellum Show a Containment Strategy for Toxic Intermediates

Clostridium thermocellum, a cellulolytic thermophilic anaerobe, is considered by many to be a prime candidate for the realization of consolidated bioprocessing (CBP) and is known as an industry standard for biofuel production. C. thermocellum is among the best biomass degraders identified to date in nature and produces ethanol as one of its main products. Many studies have helped increase ethanol titers in this microbe; however, ethanol production using C. thermocellum is still not economically viable. Therefore, a better understanding of its ethanol synthesis pathway is required. The main pathway for ethanol production in C. thermocellum involves the bifunctional aldehyde-alcohol dehydrogenase (AdhE). To better understand the function of the C. thermocellum AdhE, we used cryo-electron microscopy (cryo-EM) to obtain a 3.28 A structure of the AdhE complex. This high-resolution structure, in combination with molecular dynamics simulations, provides insight into the substrate channeling of the toxic intermediate acetaldehyde, indicates the potential role of C. thermocellum AdhE to regulate activity and cofactor pools, and establishes a basis for future engineering studies. The containment strategy found in this enzyme offers a template that could be replicated in other systems where toxic intermediates need to be sequestered to increase the production of valuable biochemicals.

09 BIOMASS FUELS↗

DOE OSTI2026

Pathways of bio-jet adoption in the US aviation industry with implications for the overall transportation and energy sectors: an integrated, multi-sectoral analysis of future scenarios

Bio-jet adoption has emerged as an attractive option to complement and supplement the use of refined fossil liquid fuels in the aviation industry in the US. However, there are significant uncertainties surrounding the costs of bio-jet including but not limited to costs of feedstock, transformation costs and the competition with co-products of bio-jet that may be demanded elsewhere in the transportation or energy sectors. This study models alternative trajectories of bio-jet adoption in the US aviation industry by 2050 through the use of a global integrated multi sector dynamics model. Three bio-jet production and consumption pathways are presented- soybean oil to jet, corn ethanol to jet (ETJ) and Fisher–Tropsch-based bio-jet, with each pathway explicitly considering the co-production of renewable diesel and renewable gasoline alongside the bio-jet. Without explicit actions or technology changes to offset the technology cost of bio-jet, scenarios where bio-jet displaces refined liquids result in higher aviation fuel prices (ranging from a 25% increase to 120% increase by mid-century) and lower demand (ranging from −14% to −43%). Corn ethanol will play an important role in the US if large scale amounts of bio-jet are to be produced with smaller effects on demand and prices. While scenarios with high levels of bio-jet availability without the availability of ETJ in the US can significantly reduce emissions in the aviation sector, these reductions are achieved more through the reduction in overall aviation fuel demand rather than technology adoption.

09 BIOMASS FUELS↗

DOE OSTI2022

Pathways for the Formation of C 2+ Products under Alkaline Conditions during the Electrochemical Reduction of CO 2

Establishing how Cu facilitates the electrochemical CO 2 reduction reaction (CO2RR) to C 2+ products remains a critical challenge. Under typical reaction conditions, the pH near the electrode is considerably more alkaline than that in the bulk due to mass transport limitations. Challenges with probing alkaline pathways using computational methods have limited understanding of the CO2RR under experimentally relevant conditions. In this work, using the Volmer reaction on Cu (100), we demonstrate that predicted activation barriers can substantially differ between acidic and alkaline pathways. We compute reaction energetics for alkaline *CO protonation and find that, while the formation of *CHO is preferred thermodynamically, the formation of *COH is favored kinetically at high overpotential. However, we find formation of *CHO via reaction of *H and *CO feasible at room temperature. Further, we report potential-dependent energetics for forming the first C-C bond in CO2RR and find that CO dimerization likely dominates. Finally, we investigate how long-range van der Waals interactions impact our results by comparing to the meta-GGA B97M-rV.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2023

Pinoresinol rescues developmental phenotypes of Arabidopsis phenylpropanoid mutants overexpressing FERULATE 5-HYDROXYLASE

Most phenylpropanoid pathway flux is directed toward the production of monolignols, but this pathway also generates multiple bioactive metabolites. The monolignols coniferyl and sinapyl alcohol polymerize to form guaiacyl (G) and syringyl (S) units in lignin, components that are characteristic of plant secondary cell walls. Lignin negatively impacts the saccharification potential of lignocellulosic biomass. Although manipulation of its content and composition through genetic engineering has reduced biomass recalcitrance, in some cases, these genetic manipulations lead to impaired growth. Here, the reduced-growth phenotype is often attributed to poor water transport due to xylem collapse in low-lignin mutants, but alternative models suggest that it could be caused by the hyper- or hypoaccumulation of phenylpropanoid intermediates. In Arabidopsis thaliana, overexpression of FERULATE 5-HYDROXYLASE (F5H) shifts the normal G/S lignin ratio to nearly pure S lignin and does not result in substantial changes to plant growth. In contrast, when we overexpressed F5H in the low-lignin mutants cinnamyl dehydrogenase c and d (cadc cadd), cinnamoyl-CoA reductase 1, and reduced epidermal fluorescence 3, plant growth was severely compromised. In addition, cadc cadd plants overexpressing F5H exhibited defects in lateral root development. Exogenous coniferyl alcohol (CA) and its dimeric coupling product, pinoresinol, rescue these phenotypes. These data suggest that mutations in the phenylpropanoid pathway limit the biosynthesis of pinoresinol, and this effect is exacerbated by overexpression of F5H, which further draws down cellular pools of its precursor, CA. Overall, these genetic manipulations appear to restrict the synthesis of pinoresinol or a downstream metabolite that is necessary for plant growth.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Computationally evaluating high-yield metabolites for sustainable aviation fuel (SAF) using machine learning

The computational tool described in this report helps identify promising biological pathways that produce SAF platform molecules (either a drop-in SAF, or a precursor that can be easily converted to a drop-in SAF). The workflow the computational tool follows first identifies possible biological pathways from a user-defined metabolite. These pathways may, or may not lead to a SAF platform molecule, thus the second step involves insilico testing of the end product of each pathway to assess whether it is, or is not, a SAF platform molecule. The identification of biological pathways performed in the first step is facilitated by linking the metabolite to a biological reaction database. Pathways are found by identifying pathways in the reaction database that include the metabolite. The computational tool includes an alternative way to find pathways. The alternative way develops a Flux Balanced Analysis (FBA), and modifying the FBA to include reactions that transform the metabolite. These modifications serve as a basis for understanding, in a semi-quantitative way, if there is an increase in the flux to desirable products. The second step, in silico testing of the end-products, is accomplished by estimating key physical properties relevant to SAF. When good models are available, we have integrated those models into the computational tool. In a few instances, we have developed our own models. In all instances, we have validated the models against available measured data. Finally, we have evaluated the effectiveness of our computational tool by genetically engineering Rhodosporidium toruloides. Validation occurred without the use of a FBA, and further validation is required.

09 BIOMASS FUELS↗

DOE OSTI2021

A systems engineering framework for the optimization of food supply chains under circular economy considerations

The current linear “take-make-waste-extractive” model leads to the depletion of natural resources and environmental degradation. Circular Economy (CE) aims to address these impacts by building supply chains that are restorative, regenerative, and environmentally benign. This can be achieved through the re-utilization of products and materials, the extensive usage of renewable energy sources, and ultimately by closing any open material loops. Such a transition towards environmental, economic and social advancements requires analytical tools for quantitative evaluation of the alternative pathways. Here, in this work, we present a novel CE system engineering framework and decision-making tool for the modeling and optimization of food supply chains. First, the alternative pathways for the production of the desired product and the valorization of wastes and by-products are identified. Then, a Resource-Task-Network representation that captures all these pathways is utilized, based on which a mixed-integer linear programming model is developed. This approach allows the holistic modeling and optimization of the entire food supply chain, taking into account any of its special characteristics, potential constraints as well as different objectives. Considering that typically CE introduces multiple, often conflicting objectives, we deploy here a multi-objective optimization strategy for trade-off analysis. A representative case study for the supply chain of coffee is discussed, illustrating the steps and the applicability of the framework. Single and multi-objective optimization formulations under five different coffee-product demand scenarios are presented. The production of instant coffee as the only final product is shown to be the least energy and environmental efficient scenario. On the contrary, the production solely of whole beans sets a hypothetical upper bound on the optimal energy and environmental utilization. In both problems presented, the amount of energy generated is significant due to the utilization of waste generated for the production of excess energy.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

DOE OSTI2023

A dynamic kinetic model captures cell-free metabolism for improved butanol production

Cell-free systems are useful tools for prototyping metabolic pathways and optimizing the production of various bioproducts. Mechanistically-based kinetic models are uniquely suited to analyze dynamic experimental data collected from cell-free systems and provide vital qualitative insight. However, to date, dynamic kinetic models have not been applied with rigorous biological constraints or trained on adequate experimental data to the degree that they would give high confidence in predictions and broadly demonstrate the potential for widespread use of such kinetic models. In this work, we construct a large-scale dynamic model of cell-free metabolism with the goal of understanding and optimizing butanol production in a cell-free system. Using a combination of parameterization methods, the resultant model captures experimental metabolite measurements across two experimental conditions for nine metabolites at timepoints between 0 and 24 hours. Here, we present analysis of the model predictions, provide recommendations for butanol optimization, and identify the aldehyde/alcohol dehydrogenase as the primary bottleneck in butanol production. Sensitivity analysis further reveals the extent to which various parameters are constrained, and our approach for probing valid parameter ranges can be applied to other modeling efforts.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2022

HydroGEN Overview: A Consortium on Advanced Water Splitting Materials

advanced water splitting technologies↗

DOE OSTI2023

HydroGEN and H2NEW Data Hub

HydroGEN is a multi-lab consortium focused on early-stage R&D in H2 production, supported by the U.S. Department of Energy (DOE), Office of Energy Efficiency and Renewable Energy (EERE), Hydrogen and Fuel Cell Technologies Office (HFTO). The consortium advances research and development (R&D) of innovative materials for advanced water splitting (AWS) technologies to enable clean, sustainable and low-cost ($1/kg H2) hydrogen production and fosters cross-cutting innovation using theory-guided applied materials R&D to advance all emerging water-splitting pathways for hydrogen production. Hydrogen from Next-generation Electrolyzers of Water (H2NEW) is a consortium of nine U.S. Department of Energy (DOE) national laboratories focused on making large-scale electrolyzers, which produce hydrogen from electricity and water, more durable, efficient, and affordable. The presentation introduces the HydroGEN data hub to the H2NEW consortium and encourages H2NEW members to use it to store and share data with team members and eventually the public. The two consortia share this data hub.

advanced water splitting technologies↗

DOE OSTI2023

HydroGEN Overview: A Consortium on Advanced Water Splitting Materials

HydroGEN is a multi-lab consortium focused on early-stage R&D in H2 production, supported by the U.S. Department of Energy (DOE), Office of Energy Efficiency and Renewable Energy (EERE), Hydrogen and Fuel Cell Technologies Office (HFTO). The consortium advances research and development (R&D) of innovative materials for advanced water splitting (AWS) technologies to enable clean, sustainable and low-cost ($1/kg H2) hydrogen production and fosters cross-cutting innovation using theory-guided applied materials R&D to advance all emerging water-splitting pathways for hydrogen production. HydroGEN is focused on low technology readiness level AWS technologies, including low- (alkaline exchanged membrane electrolysis) and high-temperature electrolysis (proton-conducting solid oxide electrolysis), photoelectrochecmical (PEC) and solar thermochemical (STCH) water splitting. This presentation will provide an overview of the HydroGEN EMN and technical highlights of a few lab-led and FOA-awarded R&D projects. HydroGEN continues to grow its community of industry, university, and national laboratories, forming a national innovation ecosystem focused on renewable hydrogen production.

advanced water splitting materials↗

DOE OSTI2023

Foundational Low-Carbon Hydrogen Research

HydroGEN is a multi-lab consortium focused on early-stage R&D in H2 production, supported by the U.S. Department of Energy (DOE), Office of Energy Efficiency and Renewable Energy (EERE), Hydrogen and Fuel Cell Technologies Office (HFTO). The consortium advances research and development (R&D) of innovative materials for advanced water splitting (AWS) technologies to enable clean, sustainable and low-cost ($1/kg H2) hydrogen production and fosters cross-cutting innovation using theory-guided applied materials R&D to advance all emerging water-splitting pathways for hydrogen production. HydroGEN is focused on low technology readiness level AWS technologies, including low- (alkaline exchanged membrane electrolysis) and high-temperature electrolysis (proton-conducting solid oxide electrolysis), photoelectrochecmical (PEC) and solar thermochemical (STCH) water splitting. This presentation will provide an overview of the HydroGEN EMN, its relationship to the H2NEW consortium, technical highlights of a few lab-led R&D projects, and the community appproach to benchmarking and protocol development for advanced water splitting technologies. Low-carbon hydrogen plays a vital role in the carbon dioxide utilization and decarbonization of several chemical and fuel industries.

advanced water splitting materials AEM↗

DOE OSTI2026

Simulating competition in the US bioeconomy to produce hard‐to‐electrify transportation fuels using limited biomass resources

This study presents a novel bioeconomy optimization framework, BiOpt, designed to address critical questions regarding the strategic use of limited US biomass resources for biofuel production. By integrating detailed techno-economic analyses, life cycle assessments, and resource assessment data, BiOpt optimizes resource distributions across competing technologies to maximize economic performance and/or minimize greenhouse gas emissions. Using feedstock scenarios from the 2023 Billion Ton Study, the analysis explores optimal biomass allocations across sustainable aviation fuel, diesel, and marine biofuel conversion pathways given varying production targets and policy incentives. Results demonstrate distinct feedstock preferences and pathway utilizations when prioritizing economic returns vs. emissions reductions. For instance, fats, oils, and greases were highly favored in cost-optimized scenarios, while low-carbon feedstocks such as wet waste dominated greenhouse gas-minimized strategies. The findings underscore the pivotal role of policy incentives and technological advances in shaping biofuel supply chains and provide actionable insights for scaling sustainable biofuel production to decarbonize hard-to-electrify sectors. This framework offers a robust tool for policymakers and stakeholders to evaluate biofuel strategies that balance energy output, economic viability, and environmental impact.

09 BIOMASS FUELS↗

DOE OSTI2026

Industrial deposition of wavelength-shifting films for liquid argon photon detection systems

The Deep Underground Neutrino Experiment (DUNE) Phase-II Far Detector is considering an approximately 2000 m 2 photon detection system to achieve a target mean light yield of 180 PE/MeV. Meeting this requirement demands scalable, cost-effective, and high-quality wavelength-shifter (WLS) coatings capable of converting 127 nm liquid-argon scintillation light into visible photons with controlled and reproducible optical performance. We report on the successful realization of an industrial physical vapor deposition (PVD) process forp-terphenyl (pTP) coatings, adapted from vacuum deposition techniques developed for OLED display manufacturing, to produce uniform WLS layers on large-area inorganic substrates, a task traditionally challenged by adhesion and uniformity issues at organic-inorganic interfaces. Surface characterization by profilometry and spectroscopic measurements demonstrates edge-region thickness variation below 10% and emission spectra consistent with high-quality pTP reference samples. The industrial process demonstrates reproducibility, scalability, and significantly reduced production time compared to laboratory-based methods, while maintaining optical characteristics consistent with established pTP reference samples. These results establish a viable pathway for mass production of high-performance pTP coatings for DUNE FD3 and future neutrino experiments, from a coating manufacturing and process standpoint. Detector-level performance validation, including quantitative VUV conversion efficiency measurements at 127 nm, is identified as future work.

47 OTHER INSTRUMENTATION↗

DOE OSTI2022

Fe Oxidation and Species Distribution at the Rock–Fluid Interface of Marcellus Shale Reacted with Hydraulic Fracturing Fluid

Hydraulic fracturing of shale reservoirs resulted in significant opportunity for increased oil and gas production in the United States. Rock-fluid interactions can cause mineral dissolution and precipitation reactions that lead to permeability changes in the shale matrix, which ultimately may affect transport pathways and hydrocarbon production. Understanding the distribution of secondary precipitates, such as barite and Fe(III) (hydro)oxides, and cation leaching at the rock-fluid interface is an important step to further investigate how these geochemical processes can change permeability and transport pathways. In this study, thin sections of the fracture-matrix interface were made from reacted Marcellus shale cores. The thin sections were characterized using synchrotron X-ray fluorescence imaging and synchrotron X-ray absorption spectroscopy. Fe species with different oxidation states were identified in the maps, together with barite and Ca distribution. The results show that ferrihydrite, as newly formed Fe(III)-bearing precipitates, aligned well with the border of the Ca (e.g., calcite) leaching region in the reaction front. Some Fe-containing clay also dissolved, but the dissolution region for the clay was not as deep as the calcite. Further, the reaction front is about three times deeper in the direction parallel to the shale bedding than that perpendicular to the bedding. The Ca leaching region can be an index for reaction front detection for Marcellus shale. Reactive transport modeling was conducted and the predicted Ca leaching boarder align well with ferrihydrite precipitation, consistent with the experimental observation. The carbonate mineral dissolution can be crucial to promote fluid access into the shale matrix. Together with our previous study on the shale reactive surface, this follow-up study showed similar Ca leaching region and Fe(III) precipitates distribution in the reaction front regardless of barite precipitation on the surface, indicating that the barite coatings on the surface may not pose a significant impact on reactive transport at the shale-fluid interface.

04 OIL SHALES AND TAR SANDS↗

DOE OSTI2026

Acetate-based biological platforms: Bridging carbon dioxide utilization and high-value bioproduct production in oleaginous yeasts

Acetate is emerging as a promising two-carbon substrate in the circular bioeconomy, bridging the gap between single-carbon sources and high-value biofuels and bioproducts. This review examines the key pathways for acetate production, including the electrochemical reduction of carbon dioxide, syngas fermentation, and biological acetogenesis. It focuses on acetate metabolism in oleaginous yeasts, such as Yarrowia lipolytica and Rhodotorula toruloides, which efficiently convert acetate-derived acetyl-CoA units into diverse bioproducts such as lipids, fatty alcohols, triacetic acid lactone, and carotenoids. Recent advances in metabolic engineering, transcriptomics, and metabolic flux analysis have improved the understanding of acetate assimilation in these organisms, thereby increasing their potential for industrial applications. In addition, the feasibility of a biological gas-to-liquid platform that utilizes acetate as a central intermediate for scalable biomanufacturing is discussed. Integrating acetate utilization with sustainable production strategies offers a promising path to advance the bio-based economy. Using acetate as a versatile metabolic intermediate enables the conversion of industrial emissions into biofuels and bioproducts while avoiding the energetic and toxicity constraints associated with direct fermentation of gaseous substrates.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Operating Strategies for Dispatchable PEM Electrolyzers that Enable Low-Cost Hydrogen Production

Hydrogen is a pathway to enabling decarbonization across multiple economic sectors that cannot be directly decarbonized with electricity including heat for industrial operations, medium- and heavy-duty transportation applications, long-duration energy storage, and as a feedstock for chemical synthesis. Producing hydrogen at low costs and carbon footprint is likely essential to economically decarbonize these otherwise "hard to decarbonize" sectors. Low-temperature polymer electrolyte membrane (PEM) electrolysis produces hydrogen from water and electricity and is a rapidly developing pathway towards making hydrogen at the scales required for decarbonization applications. The levelized cost of electrolytic hydrogen is dependent on the capital cost, efficiency, and durability of the electrolyzer system as well as the price of electricity supplied to the electrolyzer and the annual utilization of the electrolyzer (capacity factor). Electricity price and capacity factor depend on the source of energy that the system uses and impact other economic factors. Electricity price and capacity factor and the connections between other aspects of hydrogen production via PEM electrolyzers are the focus of this work. PEM electrolyzers have conventionally been operated at high capacity factors using electricity purchased from utilities with a constant price throughout the year. To achieve lower effective electricity prices, recent work has investigated opportunities for electrolyzers to purchase electricity in wholesale markets, where the cost of electricity varies hourly. This option can lead to lower electricity costs when the electrolyzer is a controllable load that ramps up and down rapidly and turns on and off frequently. This configuration and operating strategy capitalizes on times of low wholesale electricity prices and results in lower hydrogen levelized costs than constant operation due to reduced electricity costs even though the reduced capacity factor increases the cost of recovering the capital investment. Cycling on and off frequently also has implications for electrolyzer durability. An electrolyzer configuration where it is directly connected to renewable generation such as wind or solar, only running when the generator is producing energy, has similar implications on operating strategy, hydrogen levelized cost, capacity factor, and durability. This work provides insight into the relationships between dispatchable electrolyzer operating strategies and the cost of producing hydrogen from these systems, outlining strategies and opportunities to minimize production costs while minimizing operations that are likely to negatively impact system durability and efficiency. We find strategies that minimize electrolyzer cycling and the resulting durability impacts while increasing the hydrogen levelized cost only slightly above the minimum. We also find opportunities for batteries to minimize the number of cycles in systems directly connected to renewable generation. These findings outline key opportunities for future electrolyzer deployments and the synergistic benefits between electrolyzers and increased deployment of renewable energy generation like wind and solar. They also inform research and development that is reducing electrolyzer capital cost while managing durability impacts.

electricity markets↗

DOE OSTI2022

Controlling Selectivity in Plasmonic Catalysis: Switching Reaction Pathway from Hydrogenation to Homocoupling Under Visible‐Light Irradiation

Abstract Plasmonic catalysis enables the use of light to accelerate molecular transformations. Its application to the control reaction selectivity is highly attractive but remains challenging. Here, we have found that the plasmonic properties in AgPd nanoparticles allowed different reaction pathways for tunable product formation under visible‐light irradiation. By employing the hydrogenation of phenylacetylene as a model transformation, we demonstrate that visible‐light irradiation can be employed to steer the reaction pathway from hydrogenation to homocoupling. Our data showed that the decrease in the concentration of H species at the surface due to plasmon‐enhanced H 2 desorption led to the control in selectivity. These results provide important insights into the understanding of reaction selectivity with light, paving the way for the application of plasmonic catalysis to the synthesis of 1,3‐diynes, and bringing the vision of light‐driven transformations with target selectivity one step closer to reality.

Peiris, Erandi↗

DOE OSTI2020

Multiphase Reacting Flow Simulations and Optimization of Commercial-Scale Aerobic Bioreactors

Aerobic bioreactors are widely used in the synthesis of value-added products in pharmaceutical and biotechnology industries, wherein microbial bioreactions aid in the conversion of sugars to macromolecules. These reactors are also being actively investigated as a cost-effective pathway for the production of low-value commodities such as biofuels and animal feed. Reactor scale-up is one of the important challenges when designing these systems. Gas and liquid phase transport, mass-transfer, and mixing physics at large length scales can significantly affect microbial conversion rates, particularly when the microbial reaction requires a narrow set of conditions. These phenomena are difficult to study in small-scale bench-top reactors that are typically well-mixed. Predictive computational fluid dynamics (CFD) based simulations can therefore aid in the design and optimization of these reactors. This work presents multiphase Euler-Euler CFD simulations of commercial-scale (~ 500 m3) aerobic bioreactors. Our mathematical model treats the gas and liquid as interpenetrating phases. This approach reduces the computational complexity of tracking individual gas bubbles that are several orders of magnitude smaller than reactor dimensions. We solve the Reynolds averaged Navier-Stokes (RANS) multiphase equations that account for phase and chemical species transport, interphase mass and momentum transfer and uses a phenomenological model for oxygen uptake by microbes. We use a customized solver derived from open-source CFD toolbox, OpenFOAM, to perform these simulations, which has been validated against small-scale reactors in our previous work. This work examines the performance of three different reactor designs, viz. bubble column reactor, airlift reactor with an internal draft tube, and a stirred-tank reactor with Rushton impellers. Reactor oxygen mass-transfer coefficient, gas hold-up, and oxygen distribution are critically analyzed among reactors, and sensitivity studies pertaining to gas flow rates and reactor geometry will be presented.

09 BIOMASS FUELS↗