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At least 523 records · Page 29

Genetic tools to optimize lignocellulose conversion in anaerobic fungi and interrogate their genomes. Final Report

The project advances the development of anaerobic fungi, emerging model organisms for the deconstruction and conversion of untreated lignocellulosic feedstocks. Anaerobic fungi are native to the digestive tracts of large ruminants that are central to lignocellulose degradation, degrading more than 50% of the biomass by weight with similar efficiency regardless of lignin composition, which can inhibit other fungi.

09 BIOMASS FUELS↗

Elemental Analysis of Bio-Oils by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). Laboratory Analytical Procedure (LAP), Issue Date: May 13, 2022

Concentrations of inorganic elements is a key quality metric for bio-oils as certain elements impact upgrading processes and product quality. Unless reduced or removed during production and processing, alkali and alkaline metals native to lignocellulosic biomass can carry over into bio-oils contributing to ash content and degraded catalyst performance during upgrading to hydrocarbon fuels or chemical products. Non-metallic elements such as sulfur and phosphorus can also negatively impact upgrading catalysts and product quality. Inductively coupled plasma optical emission spectroscopy (ICP-OES) can be used to measure inorganic elements of interest in bio-oils. This procedure covers the preparation and analysis of fast pyrolysis (FP) and catalytic fast pyrolysis (CFP) bio-oils. The concentrations of these elements can indicate potential bio-oil quality from the perspective of deoxygenation processes. The implications of elemental composition will depend on process parameters such as upgrading catalyst sensitivities. This Laboratory Analytical Procedure (LAP) covers two methods for quantification of inorganic elements by ICP-OES: Procedure A uses microwave assisted digestion with concentrated nitric acid, and Procedure B is an organic ICP-OES method utilizing a diglyme solvent.

09 BIOMASS FUELS↗

Effective Biomass Fractionation through Oxygen-Enhanced Alkaline-Oxidative Pretreatment

The high recalcitrance of plant cell walls is an obstacle for effective chemical or biological conversion into renewable chemicals and transportation fuels. Here, we investigated the utilization of both oxygen (O 2 ) and hydrogen peroxide (H 2 O 2 ) as co-oxidants during alkalineoxidative pretreatment to improve biomass fractionation and increase enzymatic digestibility. The oxidative pretreatment of hybrid poplar was studied over a variety of conditions. Employing O 2 in addition to H 2 O 2 as a co-oxidant during the two-stage alkaline pre-extraction/coppercatalyzed alkaline hydrogen peroxide (Cu-AHP) pretreatment process resulted in a substantial improvement in delignification relative to using H 2 O 2 alone during the second-stage Cu-AHP pretreatment, leading to high overall sugar yields even at H 2 O 2 loadings as low as 2% (w/w of original biomass). The presence of H 2 O 2 , however, was both critical and synergistic. Performing analogous reactions in the absence of H 2 O 2 resulted in approximately 25% less delignification and a 30% decrease in sugar yields. The lignin isolated from this dual oxidant second stage had high aliphatic hydroxyl group content and reactivity to isocyanate, indicating that it is a promising substrate for the production of polyurethanes. To test the suitability of the isolated lignin as a source of aromatic monomers, the lignin was subjected to a sequential Bobbitt’s salt oxidation followed by formic-acid catalyzed depolymerization process. Monomer yields of approximately 17% (w/w) were obtained, and the difference in yields was not significant between lignin isolated from our Cu-AHP process with and without O 2 as a co-oxidant. Thus, the addition of O 2 did not lead to significant lignin crosslinking, a result consistent with the twodimensional heteronuclear single-quantum coherence (2D HSQC) NMR spectra of the isolated lignin.

09 BIOMASS FUELS↗

Testing of the Defense Waste Processing Facility Cold Chemical Dissolution Method in Sludge Batch 10 Qualification

For each sludge batch that is processed in the Defense Waste Processing Facility (DWPF), the Savannah River National Laboratory (SRNL) tests the applicability of the digestion methods used by the DWPF Laboratory for elemental analysis of Sludge Receipt and Adjustment Tank (SRAT) Receipt samples and SRAT Product process control samples. DWPF SRAT samples are typically dissolved using a method referred to as the DWPF Cold Chemical or Cold Chem Method (CC), (see DWPF Procedure SW4-15.201- 2.1).

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Decarbonization of Wastewater Treatment with Microalgae Processes.

Microalgae can contribute to the decarbonization of wastewater treatment by municipalities, industries and agriculture, by substituting sunlight for energy intensive conventional treatment processes, while capturing nutrients and carbon in the algal biomass. The carbon fixed into the algal biomass can be converted to renewable natural gas (RNG) using anaerobic digestion or into renewable diesel and sustainable aviation fuel (SAF) using hydrothermal liquefaction (HTL), with greatly reduce the carbon footprints compared to conventional fuels. Bioplastics, biofertilizers and other bioproducts from microalgae biomass also provide potential benefits in greenhouse gas (GHG) reduction and domestic supply chains. Microalgae technologies can counter eutrophication caused by harmful algal blooms by recovering nutrients, N and P, from wastewaters, and even from very low nutrient concentrations found in surface waters. CO2 is required in microalgae cultivation and wastewater treatment to support maximal rates of the photosynthesis, with the CO2 obtained from the wastes themselves, other local sources or even atmospheric CO2. Greenhouse gas mitigation with microalgae processes is based on life cycle assessments, comparing such green processes with current energy intensive wastewater treatment, and nutrient reduction technologies. Microalgae technologies are specifically relevant to smaller, often disadvantaged, communities, where currently about 5,000 algae wastewater treatment ponds are operated by public utilities with many more are operated by industries. However, many, if not most, of these pond facilities require urgent technology upgrades to achieve the potential and goals of low cost GHG mitigation and efficient nutrient recycling. MicroBio Engineering Inc. is developing and has demonstrated several technologies that combine innovative carbon mitigation and decarbonization technologies for a circular economy, with longer-term potential for large-scale biofuels and biofertilizer production.

09 BIOMASS FUELS↗

Decarbonization of Wastewater Treatment with Microalgae Processes.

Microalgae can contribute to the decarbonization of wastewater treatment by municipalities, industries and agriculture, by substituting sunlight for energy intensive conventional treatment processes, while capturing nutrients and carbon in the algal biomass. The carbon fixed into the algal biomass can be converted to renewable natural gas (RNG) using anaerobic digestion or into renewable diesel and sustainable aviation fuel (SAF) using hydrothermal liquefaction (HTL), with greatly reduce the carbon footprints compared to conventional fuels. Bioplastics, biofertilizers and other bioproducts from microalgae biomass also provide potential benefits in greenhouse gas (GHG) reduction and domestic supply chains. Microalgae technologies can counter eutrophication caused by harmful algal blooms by recovering nutrients, N and P, from wastewaters, and even from very low nutrient concentrations found in surface waters. CO2 is required in microalgae cultivation and wastewater treatment to support maximal rates of the photosynthesis, with the CO2 obtained from the wastes themselves, other local sources or even atmospheric CO2. Greenhouse gas mitigation with microalgae processes is based on life cycle assessments, comparing such green processes with current energy intensive wastewater treatment, and nutrient reduction technologies. Microalgae technologies are specifically relevant to smaller, often disadvantaged, communities, where currently about 5,000 algae wastewater treatment ponds are operated by public utilities with many more are operated by industries. However, many, if not most, of these pond facilities require urgent technology upgrades to achieve the potential and goals of low cost GHG mitigation and efficient nutrient recycling. MicroBio Engineering Inc. is developing and has demonstrated several technologies that combine innovative carbon mitigation and decarbonization technologies for a circular economy, with longer-term potential for large-scale biofuels and biofertilizer production.

09 BIOMASS FUELS↗

Decarbonization of Wastewater Treatment with Microalgae Processes.

Microalgae can contribute to the decarbonization of wastewater treatment by municipalities, industries and agriculture, by substituting sunlight for energy intensive conventional treatment processes, while capturing nutrients and carbon in the algal biomass. The carbon fixed into the algal biomass can be converted to renewable natural gas (RNG) using anaerobic digestion or into renewable diesel and sustainable aviation fuel (SAF) using hydrothermal liquefaction (HTL), with greatly reduce the carbon footprints compared to conventional fuels. Bioplastics, biofertilizers and other bioproducts from microalgae biomass also provide potential benefits in greenhouse gas (GHG) reduction and domestic supply chains. Microalgae technologies can counter eutrophication caused by harmful algal blooms by recovering nutrients, N and P, from wastewaters, and even from very low nutrient concentrations found in surface waters. CO2 is required in microalgae cultivation and wastewater treatment to support maximal rates of the photosynthesis, with the CO2 obtained from the wastes themselves, other local sources or even atmospheric CO2. Greenhouse gas mitigation with microalgae processes is based on life cycle assessments, comparing such green processes with current energy intensive wastewater treatment, and nutrient reduction technologies. Microalgae technologies are specifically relevant to smaller, often disadvantaged, communities, where currently about 5,000 algae wastewater treatment ponds are operated by public utilities with many more are operated by industries. However, many, if not most, of these pond facilities require urgent technology upgrades to achieve the potential and goals of low cost GHG mitigation and efficient nutrient recycling. MicroBio Engineering Inc. is developing and has demonstrated several technologies that combine innovative carbon mitigation and decarbonization technologies for a circular economy, with longer-term potential for large-scale biofuels and biofertilizer production.

09 BIOMASS FUELS↗

High Solids Loading Aqueous Slurry Formation of Corn Stover Before Pretreatment in a Fed-batch Bioreactor

With the increase on population the world will depend on renewable sources to meet the increasingly energy needs. Use of lignocellulosic biomass as a renewable source has been proven efficient for conversion to cellulosic ethanol and capable of contributing to achieve the threshold on energy demand while reducing greenhouse gases in 90% when compared with fossil fuels (Wang et al., 2007, 2012). However, the processing of biomass encounters limitations in feeding and flow within biorefineries due to the system plugging by compaction and slurry high yield stress, preventing transport of biomass materials and in some cases unexpected plant shutdowns that results in high operational costs (dos Santos et al., 2021; Ximenes et al., 2021). Different solutions for biomass handling and slurry formation from densified materials like pellets have been studied. Cellulosic biomass residues are typically processed in pretreatment reactors to which acid or base is added (Humbird et al., 2002). Other pretreatments such as liquid hot water (LHW) and steam explosion that act without the addition of chemicals use pressures above the saturation vapor of water, to disrupt biomass structure (Ruiz et al., 2021). Although these methods are widely known, the use of pretreatments like acid digestion and LHW present challenges like waste disposal and could be energy inefficient (Mosier et al., 2005). Alternatively other approaches like enzymatic liquefaction have been applied for biomass transformation into slurries with promising results and without the use of pretreatments (dos Santos et al., 2021). Nevertheless, enzymatic liquefaction implementation faces difficulties due to the recalcitrant properties of biomass, its variability, and the release of enzyme inhibitors. While it has been proven that chemical composition properties in biomass have effects on enzyme inhibition (Huang et al., 2022; Kim et al., 2011; Zhai et al., 2018), hindering the efficiency of the liquefaction, not ample research has been conducted in understanding the physical properties as particle size, porosity, water adsorption retention and their effect in the ability to form a slurry at high solids concentration (Yan et al., 2020). In order to improve lignocellulosic biomass handling and formation of biomass slurries, enzyme assisted liquefaction for slurry creation from corn stover at solids loadings up to 30% is reported in this work. Two different kinds of biomass (pelleted corn stover and cobs) were liquefied in a fed-batch process using commercial enzymes Celluclast 1.5L or Ctec-2 at 1FPU or 3 FPU per gram of dry solids in 10 mM sodium citrate buffer solution (pH 4.8). Pellets were fed into a 1 L stirred bioreactor according to a predefined fed-batch protocol over the first 5 hours until reaching 30% of solids loading. After 6, 24 and 96 hours, samples were taken and characterized with respect to their sugar composition, rheology and water absorption. Successful slurry creation with dramatically reduced yield stress was achieved for corn stover for both assessed enzymes. Yield stresses of 178±7 Pa (3 FPU, Celluclast 1.5L) and 79±6 Pa (3 FPU, Ctec-2) were measured for corn stover at 24 hours, compared to 6,000 Pa for samples without enzyme. Yield stress was 155± 29 Pa (3FPU, Ctec-2) and 257 ± 72 Pa (1 FPU, Celluclast 1.5L) for corn cobs at 24 hours. Yield stress decreased when residence time increased with an enhanced fluidity noted for higher enzyme concentrations. A profile for 6, 24 and 96h of yield stress measurements is presented.

Gutierrez, Diana↗

Final Technical Report

The Department of Energy is interested in technologies that support the sustainable production of fuels, chemicals, and other bioproducts from plant biomass, to offset the nation’s reliance on fossil resources. The plant cell wall of energy crops provides the largest reservoir of raw materials for bioproducts. However, the widespread use of plant cell walls is hampered by their complexity and resistance to breakdown. To improve the productivity and cost-effectiveness of using energy crops to generate bioproducts, the fundamental problem of deconstructing plant cell walls must be addressed. This project developed and evaluated an innovative genetic modification technology to produce strategically designed enzymes that specifically accumulate in the plant cell wall. The resulting enzyme-engineered energy crops are expected to grow normally under natural conditions but break down more quickly and easily under high temperature during the production of biobased products. As such, this plant cell wall targeting enzyme engineering effort will reduce the cost of plant cell wall deconstruction and ultimately improve the economics of bioproducts. The overall objective of this project is to develop and evaluate the in-planta enzyme engineering technology to reduce lignocellulose deconstruction cost. The concept was first validated using tobacco plant, a model plant system that is typically used in lab testing for initial concept validation. Then the enzyme optimization was validated using switchgrass, the energy crop to be used to produce bioproducts. There are three specific objectives in this Phase I project: (1) validate the enzyme optimization concept using tobacco plant, a model plant system. (2) validate the enzyme optimization concept using switchgrass. (3) techno-economic analysis (TEA) for further scale-up application. By the end of this project, in-planta enzyme engineering was validated in both tobacco and switchgrass plants, with improved enzyme activity and saccharification efficiency. The in-planta enzyme engineering in Tabacco didn’t have a significant impact on plant growth and development. Transgenic tobacco plants with in-planta cellulose degrading enzymes showed higher biomass digestibility than wild type. Gene construction and transformation in switchgrass was much longer than expected, which delayed the research progress. Besides, in-planta engineering of lignin degrading enzyme is more challenging than cellulose degrading enzyme, in terms of expression detection. Expression of lignin degrading enzyme and cellulose degrading enzyme improved biomass yield and saccharification efficiency of switchgrass, respectively. It is promising to express both genes in switchgrass for optimized overall performance. According to the results of TEA, switchgrass biomass production cost is mainly attributed to by fertility and harvesting. Biomass production profit can increase up to 10-fold depending on biomass price. The PHA production profit is also sensitive to the biomass price. The proposed technology could potentially reduce the biomass deconstruction cost from 33% to 9% of PHA revenue, making the biomass-based PHA competitive to petroleum-based polymers even in case of relatively high biomass price of biomass. Therefore, cultivation of the genetically engineered self-deconstruction switchgrass for Polyhydroxyalkanoate (PHA) production could benefit switchgrass grower and PHA producer with attractive profits for both sectors. This new enzyme optimization approach will be beneficial for bioindustries that use energy crops as feedstocks. It will improve the economic viability of converting energy crops to renewable products that support a sustainable society and helps address the Nation’s long-term strategic needs for renewable products and reduction of reliance on fossil resources.

42 ENGINEERING↗

Tank Closure Cesium Removal Ion Exchange Column CST Characterization: SRNL-STI-2024-00072 (Rev.0) pdf

Savannah River Mission Completion (SRMC) recently suspended operation of the Tank Closure Cesium Removal (TCCR) system which utilized an ion exchange (IX) process to remove radioactive cesium from waste supernate. During the demonstration phase, a total of three batches of Tank 10H dissolved salt waste were processed through the original TCCR columns. Subsequently, dissolved salt waste from Tank 9H was processed through TCCR in a phase referred to as TCCR 1A with four new IX columns. All eight columns have now been moved to Interim Safe Storage (ISS). For each batch processed through the TCCR IX columns, SRNL performed a number of analyses on surface and variable depth samples as well as batch equilibrium contact tests (BECT) to determine equilibrium loadings of Cs on the crystalline silicotitanate (CST) IX media. The BECTs were performed by lowering “teabags” containing ~0.1 g of CST each into the tank for a period of at least 10 days. The teabags were then retrieved and sent to SRNL for digestion and analysis.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

A Geodatabase Designed to Inform and Support Safe CO2 Transport-Route Planning

The National Energy Technology Laboratory developed the Carbon Capture and Storage (CCS) Pipeline Route Planning Database to inform safe and sustainable CO2 transport planning in support of decarbonization efforts. This comprehensive Esri Geodatabase contains over 90 GBs of data and 60+ spatial layers representing key considerations including natural hazards, infrastructure, energy, and social justice. Leveraging ArcGIS Pro, nationwide raster, and vector datasets containing millions of features were processed to be easily digestible for users and complex modeling software.

Romeo, Lucy↗

EDX ClaiMM: Digital Resources for the Critical Minerals and Materials Community

Securing critical mineral supply chains is essential for transitioning to a clean energy economy and for maintaining national security. Big-data analytics can serve as a cost-effective means of identifying new domestic critical mineral resources but only if data can be easily located and digested. Using ArcGIS Enterprise Sites, EDX ClaiMM was developed to increase the accessibility of critical minerals data, reducing time spent on data collection and integration. Hosted tools provide rapid visualization and exploration of key datasets, unlocking insights to support resource assessments.

Yesenchak, Rachel↗

Virtual Environment Platform for OT/IT Training Enhancement

TRADITIONAL TECHNIQUES OT/IT Concepts Operational Technology (OT) and Information Technology (IT) concepts can often be difficult to visualize Teaching Methods Traditional teaching methods lack the intuitive & immersive aspects of hands-on activities Caveat: Unless taught by Team B! Physical Limitations Digital Twins require existing systems/hardware to mirror UPDATED TECHNIQUES Virtual Environment By making use of a virtual environment, we can represent abstract concepts in a more approachable and digestible way Increased Engagement Students are more engaged with the activities and are more likely to retain the information they are given. New/Emerging Technologies As the system is currently growing and developing, the technologies in use, as well as those represented by the system, stay up-to-date.

Deroller, Nicholas F.↗

Laser Driven Hydrothermal Processing (LDHP) for Rapid Dissolution of Glassy Nuclear Debris

Discovered in 2009 on the SLAB laser system at LLNL, Laser-Driven Hydrothermal Processing (LDHP) both spalls and transiently dissolves a wetted surface (e.g., rock, concrete, paint) when that surface is struck by a laser beam pulse under specific conditions. Previous projects have explored the application of LDHP to expediting sample dissolution for post-detonation nuclear forensics. Early results were very promising, indicating that this method could decrease material dissolution time by a factor of 10 or more. However, the large and complicated lasers previously used are impractical for this application. Under the R^3 venture we aimed to benchmark the LDHP capabilities of turn-key commercial off-the-shelf (COTS) lasers, and to establish how this process could be integrated into an analytical workflow for nuclear forensic analysis. We demonstrated that geological rock standards could be quickly processed into fine powder by LDHP followed by complete acid digestion in 15 minutes, without elemental fractionation that would skew analytical results. We determined that the COTS laser can be used for LDHP that could potentially reduce dissolution time for solid nuclear debris samples in post detonation nuclear forensics. One downside to this approach, however, is that the laser interactions are different with every material, so extensive method validation would be required for all likely material compositions. The LDHP processing rates of several materials were tested and determined to be favorable in the beginning with significant slowdown due to particulate scattering. This could be mitigated either by stopping and centrifuging the sample at specific intervals, or through design of a continuous solvent flow through mechanism.

36 MATERIALS SCIENCE↗

Seminar 44, Innovative and New Ideas: BAS Best Practices for O&M Success Coaching Operators for Effective Use of Data

Regardless of a building’s inherent energy efficiency, changes in building operator behavior reduces building energy consumption by 14% on average. Giving operators readily available and easily digestible data regarding the performance of their HVAC systems increases the likelihood of energy saving actions. Many Building Automations Systems do not excel at creating user-friendly time series trend charts. This presentation presents examples of a solution to this issue developed under an award by the Department of Energy.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Prospecting for Critical Minerals and Rare Earth Elements from Marcellus Shale in the Western Portion of the Appalachian Basin with Non-Destructive Core Characterization

Identification of sources for domestic critical minerals and rare earth elements (CM/REE) has been deemed essential for the energy transition by the United States Department of Energy (DOE). The U.S. DOE’s National Energy Technology Laboratory’s (NETL) Geomaterials Characterization Laboratory has performed non-destructive core characterizations on energy-relevant rock cores for the past decade. During this time, NETL has published over 36 technical reports and made the associated data publicly available. Much of this work focuses on unconventional shale gas, subsurface carbon storage systems, and carbon-ore. These efforts provide cm-scale petrophysical and elemental data, photographic documentation, detailed core descriptions, and computed tomography (CT) data for each well. This provides a first phase prospecting resource for CM/REE resources and can provide a map for pin-pointing intervals and lithologies for further development. Using historical core characterization data from 12 Marcellus wells from the western portion of the Appalachian Basin, this study builds an improved understanding of the chemostratigraphy of the basin. X-ray fluorescence (XRF) and CT images were used to determine lithologic intervals and potential ore bodies for further analysis, including benchtop digestion and inductively coupled plasma mass spectrometry (ICP-MS) to better understand the CM/REE enrichments.

Paronish, Thomas J.↗

Device and Method for Parallel Measurement of Phosphoproteome and Proteome from Single Cells

We present the development of an immobilized metal affinity chromatography (IMAC) chip designed to enable nanoscale phosphopeptide enrichment within microfabricated nanowells. This novel platform leverages surface chemistry to immobilize high-density Nickel-Nitrilotriacetic Acid (Ni-NTA) molecules on nanowells, followed by applying Fe 3+ . The nanowell surface serves as a capture media to enrich phosphopeptides based on IMAC. The system's efficiency was validated using ß-casein as a model protein, demonstrating the chip’s capability to significantly enrich phosphopeptides. Future applications of this technology are anticipated to enable the detection of over 100 phosphopeptides from individual cells and more than 500 phosphopeptides from pools of 100 cells, offering exciting potential for single-cell phosphoproteomics. We will next apply an integrated proteomics workflow to perform multi-omics measurements, including single-cell isolation, protein digestion, and phosphopeptide enrichment, followed by LC-MS analysis of both the global proteome and phosphoproteome. Future research will explore the use of this technology to study phosphorylation dynamics in cancer cells, enhancing our understanding of cellular signaling and disease mechanisms.

59 BASIC BIOLOGICAL SCIENCES↗

Harnessing photoautotroph-methanotroph interactions for biogas conversion to fuels and chemicals using binary consortia (Project Final Technical Report)

Industrial, municipal, and agricultural waste streams containing stranded organic carbon represent a significant and underutilized feedstock to produce fuels and chemicals. With anaerobic digestion deployed at large scales to capture organic waste streams, over 6 million tons of biogas are available today. However, the utilization of biogas represents a significant challenge due to its low pressure and presence of contaminants such as H 2 S, ammonia, and volatile organic carbon compounds. To tap into this immense potential, effective biotechnologies that co-utilize both CO 2 and CH 4 are needed. Recent studies demonstrated that, in nature, microbial communities have developed a highly efficient way to recover energy and capture carbon from both CH 4 and CO 2 through metabolic coupling of methane oxidation to oxygenic photosynthesis. Using two synthetic methanotroph – photoautotroph (M-P) co-cultures that exhibit stable growth under a broad range of cultivation conditions, in this project we proposed to harness the interspecies interactions within these cocultures for biogas conversion to fuels and chemicals. To facilitate this overarching objective, we aim to develop experimental and computational tools to gain qualitative and quantitative understandings on the interactions and dynamics of the coculture at both systems and molecular levels, and to validate our findings through experiments and mutant development. The fundamental understanding on the interactions and dynamics of the photoautotroph-methanotroph will lay the foundation for the design and optimization of synthetic binary consortia for production of fuels and chemicals from biogas. We expect the knowledge gained from this project may be generally applicable to other cross-feeding binary consortium, and the tools developed can be adapted to study the interactions and dynamics of other multi-organism platforms.

09 BIOMASS FUELS↗