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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 415 records · Page 23

Data for Miscanthus giganteus Biolistic Transformation Using the Visible RUBY Red Marker Gene to Monitor Transformation Efficiency

Miscanthus × giganteus ( M × g ) is a high- yielding perennial C4 bioenergy crop, but genetic improvement by breeding is constrained by triploid sterility and clonal propagation. Improving genetic transformation methods for M × g would provide opportunities for advantageous trait introgression. Use of an easy to phenotype reporter gene is a promising strategy to improve transformation processes and efficiency. This study presents an efficient novel method for biolistic transformation of inflorescence- derived callus in M × g and demonstrates its efficacy using RUBY, a betalain-based noninvasive reporter that is visible throughout the transformation process. RUBY expression ( Zea mays codon optimized) was visible from callus stage through plantlet development into maturity. RUBY expressing independently transformed plants were confirmed by hygromycin phosphotransferase ELISA and by genomic PCR demonstrating that the RUBY phenotype is sufficient for screening transformants. The Zea mays codon optimized hygromycin selection marker was driven by previously established promoters for Miscanthus, ZmUBI and 2×35S, while the RUBY gene expression was controlled by a known Zea mays C4 promoter, Brachypodium UBI10, newly employed in Miscanthus. The construct containing the 2×35S promoter for hygromycin had a 15.1% transformation efficiency while the ZmUBI promoter had a 20.5% transformation efficiency. This study provides a novel, highly efficient protocol for successful biolistic transformation of M × g for stable expression. This study also demonstrates that RUBY expression can be used as a convenient and powerful monitor of transformation in ongoing and future work to engineer M × g into an improved bioproduct feedstock. **NOTE: in "TableS2_ProtocolComparison.csv", the data from row 665 to 971 should be removed.

Gene Editing↗

BETO 2021 Peer Review - Cyanobacteria Photosynthetic Energy Platform (1.3.4.301)

Cyanobacteria Photosynthetic Energy Platform (WBS 1.3.4.301) The primary challenge in algal technology is low energy conversion efficiency, currently up to a few percentages from photon energy to biomass energy. Most incoming solar energy is lost in the processes of photosynthesis, through mechanisms that are not fully understood. There is large room for improvement. This project develops cyanobacteria genetic tools to improve photosynthetic efficiency and biomass productivity through manipulation of energy regulation mechanisms. Increasing algal biomass productivity by 20% translates to $2 reduction of MFSP. Genetic manipulation is an essential tool in understanding and improving algal productivity. While genetic manipulation is challenging for most algae, model cyanobacteria, with in-house mutant library and genetic engineering toolboxes, allow for rapid hypothesis testing and transfer of lessons to other cyanobacteria and eukaryotic algae. We developed Energome (energy-ome) concept to guide novel engineering strategies and unleash unused potential in photosynthesis. Energome consists of cellular energy management mechanisms. For example, we found that cyanobacteria use futile cycles around glycogen and sucrose to dissipate ATP. By genetic modification, the model cyanobacterium Synechocystis 6803 showed higher energy levels, increased photosynthetic capacity, and 20% improvement in biomass yield in simulated outdoor light conditions.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Accelerated Short Circuiting in Anode-Free Solid-State Batteries Driven by Local Lithium Depletion

“Anode-free” solid-state batteries (SSBs), which have no anode active material, can exhibit extremely high energy density (≈1500 Wh L -1 ). However, there is a lack of understanding of the lithium plating/stripping mechanisms at initially lithium-free solid-state electrolyte (SSE) interfaces because excess lithium metal is often used. Here, it is demonstrated that commercially relevant quantities of lithium (>5 mAh cm -2 ) can be reliably plated at moderate current densities (1 mA cm -2 ) using the sulfide SSE Li 6 PS 5 Cl. Investigations of lithium plating/stripping mechanisms, in conjunction with cryo-ocused ion beam (FIB) imaging, synchrotron tomography, and phase-field modeling, reveal that the cycling stability of these cells is fundamentally limited by the nonuniform presence of lithium during stripping. Local lithium depletion causes isolated lithium regions toward the end of stripping, decreasing electrochemically active area and resulting in high local current densities and void formation. This accelerates subsequent filament growth and short circuiting compared to lithium-excess cells. Despite this degradation mode, here it is shown that anode-free cells exhibit comparable Coulombic efficiency to lithium-excess cells, and improved resistance to short circuiting is achieved by avoiding local lithium depletion through retention of thicker lithium at the interface. These new insights provide a foundation for engineering future high-energy anode-free SSBs.

25 ENERGY STORAGE↗

FLEX-FUEL MIXING CONTROLLED COMBUSTION ENABLED BY PRECHAMBER IGNITION

There is an imminent need to displace fossil diesel fuel with cleaner burning, domestically produced, renewable fuels for use in heavy-duty engines. Bioethanol is a prime candidate as it widely adopted in the U.S. as a gasoline additive ranging in volume percentage from 10% (E10) up to 85% (E85). Direct substitution of market available ethanol-gasoline blends for diesel fuel is not plausible as the stark reactivity differences would not constitute the same ignition quality nor achieve auto-ignition at all. This work focuses on the development of prechamber enabled mixing-controlled combustion (PC-MCC) as an advanced combustion strategy to facilitate reliable ignition and diffusion style combustion ethanol-gasoline fuel blends. PC-MCC involves integration of an actively fueled prechamber (PC) into a conventional compression ignition combustion system. When ignited, the PC ejects hot turbulent jets into the main combustion chamber that then interact with the direct injected fuel, prompting immediate ignition. The PC jet flames provide a robust thermal ignition source that allows the engine to operate agnostic of fuel composition, or flex-fuel. Computational fluid dynamics (CFD) modeling was used to assess critical design features of the PC while garnering insights into the ignition strategies that facilitate robust performance. A key finding was the ignition performance benefits of fuel-rich PC operation which yield exothermic jets. Based on the numerical findings, a prototype igniter was tested experimentally on both single and multi-cylinder engine platforms at a variety of operating conditions. The experimental results indicate flex-fuel PC-MCC is well capable of diesel-like combustion processes by demonstrating matched or improved gross thermal efficiencies and load variability within 2%. Fuel grade ethanol (E98) exhibited consistently lower NOx and immeasurable soot across the load space. E98 also demonstrated a significant improvement in thermal efficiency at light loads.

Zeman, Jared↗

Vehicle Localization in 3D World Coordinates Using Single Camera at Traffic Intersection

Optimizing traffic control systems at traffic intersections can reduce the network-wide fuel consumption, as well as emissions of conventional fuel-powered vehicles. While traffic signals have been controlled based on predetermined schedules, various adaptive signal control systems have recently been developed using advanced sensors such as cameras, radars, and LiDARs. Among these sensors, cameras can provide a cost-effective way to determine the number, location, type, and speed of the vehicles for better-informed decision-making at traffic intersections. In this research, a new approach for accurately determining vehicle locations near traffic intersections using a single camera is presented. For that purpose, a well-known object detection algorithm called YOLO is used to determine vehicle locations in video images captured by a traffic camera. YOLO draws a bounding box around each detected vehicle, and the vehicle location in the image coordinates is converted to the world coordinates using camera calibration data. During this process, a significant error between the center of a vehicle’s bounding box and the real center of the vehicle in the world coordinates is generated due to the angled view of the vehicles by a camera installed on a traffic light pole. As a means of mitigating this vehicle localization error, two different types of regression models are trained and applied to the centers of the bounding boxes of the camera-detected vehicles. The accuracy of the proposed approach is validated using both static camera images and live-streamed traffic video. Based on the improved vehicle localization, it is expected that more accurate traffic signal control can be made to improve the overall network-wide energy efficiency and traffic flow at traffic intersections.

47 OTHER INSTRUMENTATION↗

acCRISPR: an activity-correction method for improving the accuracy of CRISPR screens

Abstract High throughput CRISPR screens are revolutionizing the way scientists unravel the genetic underpinnings of engineered and evolved phenotypes. One of the critical challenges in accurately assessing screening outcomes is accounting for the variability in sgRNA cutting efficiency. Poorly active guides targeting genes essential to screening conditions obscure the growth defects that are expected from disrupting them. Here, we develop acCRISPR, an end-to-end pipeline that identifies essential genes in pooled CRISPR screens using sgRNA read counts obtained from next-generation sequencing. acCRISPR uses experimentally determined cutting efficiencies for each guide in the library to provide an activity correction to the screening outcomes via calculation of an optimization metric, thus determining the fitness effect of disrupted genes. CRISPR-Cas9 and -Cas12a screens were carried out in the non-conventional oleaginous yeast Yarrowia lipolytica and acCRISPR was used to determine a high-confidence set of essential genes for growth under glucose, a common carbon source used for the industrial production of oleochemicals. acCRISPR was also used in screens quantifying relative cellular fitness under high salt conditions to identify genes that were related to salt tolerance. Collectively, this work presents an experimental-computational framework for CRISPR-based functional genomics studies that may be expanded to other non-conventional organisms of interest.

59 BASIC BIOLOGICAL SCIENCES↗

Functional analysis of the methylerythritol phosphate pathway terminal enzymes IspG and IspH from Zymomonas mobilis

ABSTRACT Isoprenoids are a diverse family of compounds that are synthesized from two isomeric compounds, isopentenyl diphosphate and dimethylallyl diphosphate. In most bacteria, isoprenoids are produced from the essential methylerythritol phosphate (MEP) pathway. The terminal enzymes of the MEP pathway IspG and IspH are [4Fe-4S] cluster proteins, and in Zymomonas mobilis, the substrates of IspG and IspH accumulate in cells in response to O 2 , suggesting possible lability of their [4Fe-4S] clusters. Here, we show using complementation assays in Escherichia coli that even under anaerobic conditions, Z. mobilis IspG and IspH are not as functional as their E. coli counterparts, requiring higher levels of expression to rescue viability. A deficit of the sulfur utilization factor (SUF) Fe-S cluster biogenesis pathway did not explain the reduced function of Z. mobilis IspG and IspH since no improvement in viability was observed in E. coli expressing the Z. mobilis SUF pathway or having increased expression of the E. coli SUF pathway. Complementation of single and double mutants with various combinations of Z. mobilis and E. coli IspG and IspH indicated that optimal growth required the pairing of IspG and IspH from the same species. Furthermore, Z. mobilis IspH conferred an O 2 -sensitive growth defect to E. coli that could be partially rescued by co-expression of Z. mobilis IspG. In vitro analysis showed O 2 sensitivity of the [4Fe-4S] cluster of both Z. mobilis IspG and IspH. Altogether, our data indicate an important role of the cognate protein IspG in Z. mobilis IspH function under both aerobic and anaerobic conditions. IMPORTANCE Isoprenoids are one of the largest classes of natural products, exhibiting diversity in structure and function. They also include compounds that are essential for cellular life across the biological world. In bacteria, isoprenoids are derived from two precursors, isopentenyl diphosphate and dimethylallyl diphosphate, synthesized primarily by the methylerythritol phosphate pathway. The aerotolerant Z. mobilis has the potential for methylerythritol phosphate pathway engineering by diverting some of the glucose that is typically efficiently converted into ethanol to produce isoprenoid precursors to make bioproducts and biofuels. Our data revealed the surprising finding that Z. mobilis IspG and IspH need to be co-optimized to improve flux via the methyl erythritol phosphate pathway in part to evade the oxygen sensitivity of IspH.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Mechanically strengthened heterogeneous Sm-Co sintered magnets

Samarium-cobalt sintered magnets offer excellent magnetic properties, thermal stability, and corrosion resistance. They are used in a variety of defense and civilian applications, especially when elevated operation temperatures (e.g., 200 ºC to 550 ºC) are required. However, the utilization of these materials is restricted by their brittleness. Improving their mechanical resilience would make them more cost-effective, efficient, and robust in decarbonization and other function-related applications while reducing the pressure on critical material supply chains. In this paper, we engineer a series of novel heterogeneous microstructures, such as laminated coarse grain (CG)/fine grain (FG) and core/shell CG/FG microstructures, to produce unprecedented combinations of superior mechanical and magnetic properties without altering the chemical compositions of the magnets or common heat treatment procedures. Further, a 60% flexural strength enhancement is obtained using heterogeneous Sm 2 (CoFeCuZr) 17 sintered magnets with little impact on their magnetic properties. The mechanically robust heterogeneous Sm-Co sintered magnets have a minor (e.g., less than 4.6%) reduction in the energy product (BH) max due to a slightly reduced squareness of the demagnetization curve, with no decrease in either the remanence (B r ) or the intrinsic coercivity (H ci ). The flexural strengths of these heterogeneous Sm-Co magnets depend on the volume ratios and mean grain sizes of the FG/CG regions, as well as their microstructural architectures. The fine-grained regions act as mechanical strengthening sites, which can be strategically used when designing the magnet for different applications. This technology is highly compatible with existing magnet manufacturing processes and thus can be adopted readily by the magnet industry.

36 MATERIALS SCIENCE↗

Directed evolution of phosphite dehydrogenase to cycle noncanonical redox cofactors via universal growth selection platform

Abstract Noncanonical redox cofactors are attractive low-cost alternatives to nicotinamide adenine dinucleotide (phosphate) (NAD(P) + ) in biotransformation. However, engineering enzymes to utilize them is challenging. Here, we present a high-throughput directed evolution platform which couples cell growth to the in vivo cycling of a noncanonical cofactor, nicotinamide mononucleotide (NMN + ). We achieve this by engineering the life-essential glutathione reductase in Escherichia coli to exclusively rely on the reduced NMN + (NMNH). Using this system, we develop a phosphite dehydrogenase (PTDH) to cycle NMN + with ~147-fold improved catalytic efficiency, which translates to an industrially viable total turnover number of ~45,000 in cell-free biotransformation without requiring high cofactor concentrations. Moreover, the PTDH variants also exhibit improved activity with another structurally deviant noncanonical cofactor, 1-benzylnicotinamide (BNA + ), showcasing their broad applications. Structural modeling prediction reveals a general design principle where the mutations and the smaller, noncanonical cofactors together mimic the steric interactions of the larger, natural cofactors NAD(P) + .

59 BASIC BIOLOGICAL SCIENCES↗

Pathways to a net-zero-carbon water sector through energy-extracting wastewater technologies

The energy-consuming and carbon-intensive wastewater treatment plants could become significant energy producers and recycled organic and metallic material generators, thereby contributing to broad sustainable development goals, the circular economy, and the water-energy-sanitation-food-carbon nexus. This review provides an overview of the waste(water)-based energy-extracting technologies, their engineering performance, techno-economic feasibility, and environmental benefits. Here, we propose four crucial strategies to achieve net-zero carbon along with energy sufficiency in the water sector, including (1) improvement in process energy efficiency; (2) maximizing on-site renewable capacities and biogas upgrading; (3) harvesting energy from treated effluent; (4) a new paradigm for decentralized water-energy supply units.

54 ENVIRONMENTAL SCIENCES↗

Increasing Rubisco as a simple means to enhance photosynthesis and productivity now without lowering nitrogen use efficiency

Summary Global demand for food may rise by 60% mid‐century. A central challenge is to meet this need using less land in a changing climate. Nearly all crop carbon is assimilated through Rubisco, which is catalytically slow, reactive with oxygen, and a major component of leaf nitrogen. Developing more efficient forms of Rubisco, or engineering CO 2 concentrating mechanisms into C 3 crops to competitively repress oxygenation, are major endeavors, which could hugely increase photosynthetic productivity (≥ 60%). New technologies are bringing this closer, but improvements remain in the discovery phase and have not been reduced to practice. A simpler shorter‐term strategy that could fill this time gap, but with smaller productivity increases (c. 10%) is to increase leaf Rubisco content. This has been demonstrated in initial field trials, improving the productivity of C 3 and C 4 crops. Combining three‐dimensional leaf canopies with metabolic models infers that a 20% increase in Rubisco increases canopy photosynthesis by 14% in sugarcane (C 4 ) and 9% in soybean (C 3 ). This is consistent with observed productivity increases in rice, maize, sorghum and sugarcane. Upregulation of Rubisco is calculated not to require more nitrogen per unit yield and although achieved transgenically to date, might be achieved using gene editing to produce transgene‐free gain of function mutations or using breeding.

Plant Sciences↗

Final Technical Report for DE-SC0022206

This project developed foundational genetic, genomic, and epigenetic tools for anaerobic fungi (Neocallimastigomycota), a group of microorganisms with exceptional natural abilities to deconstruct lignocellulosic biomass. Efficient biomass deconstruction remains a major barrier to economical production of renewable fuels, chemicals, and materials from agricultural and forestry residues. The project sought to enable mechanistic studies and future engineering of anaerobic fungi by improving genomic resources, establishing methods for gene expression, and investigating epigenetic regulation of biomass-degrading pathways. Major accomplishments included generation of the first chromosome-scale genome assemblies for multiple anaerobic fungal species, providing publicly available genomic resources that support both engineering and fundamental biological research. The project established the first reproducible system for heterologous gene expression in anaerobic fungi and identified genomic features and mobile genetic elements that may support future development of stable transformation technologies. In parallel, the project demonstrated direct conversion of untreated lignocellulosic biomass into fuels and specialty chemicals through a fungal-yeast bioprocess and identified anaerobic fungal enzymes with utility for metabolic engineering. The research also revealed that epigenetic regulation plays an important role in controlling fungal gene expression and enzyme production, identifying potential strategies for enhancing biomass degradation. Collectively, this work established anaerobic fungi as a tractable emerging platform for bioenergy and biomanufacturing research, generated valuable public resources, trained the next generation of researchers, and advanced DOE-BER goals related to predictive biology, sustainable bioprocessing, and the circular bioeconomy.

Solomon, Kevin [University of Delaware] (ORCID:000↗

Feeding from the sun—Successes and prospects in bioengineering photosynthesis for food security

There is an urgent need for increased crop productivity to reduce food insecurity and improve sustainability. Photosynthesis converts sunlight energy into carbohydrates, providing the source of nearly all of humanity’s food. Photosynthesis is a key target for improvement, owing to inherent inefficiencies in the biochemical process. Over the last decade of advancements in bioengineering, strategies to increase the efficiency of photosynthesis were tested with proven enhancements to crop yields in field trials. Simple strategies like increasing the content of photosynthetic proteins have reliably increased photosynthesis and productivity in crops, as have more complex strategies such as bypassing photorespiration. While insertion of carbon-concentrating mechanisms into C3 plants remains an engineering challenge, modeling suggests that achieving that would have the greatest gain for crop improvement. This review discusses the many successes in improving photosynthesis achieved over the past decade and quantifies the potential for future engineering targets to increase crop productivity.

Long, Stephen P. [University of Illinois, Urbana, ↗

Use of novel refractory design and installation techniques for improved energy efficiency in iron and steel and other energy intensive industries

This paper describes the planned work to be performed under a United States Department of Energy funded project to bring together the key players necessary to develop and deploy new technology which could increase the thermal efficiency of the steel industry as well as other energy intensive industries. It is hoped that such improvements in energy efficiency will reduce the overall energy and environmental footprint of domestic industry, as well as provide economic benefit to the individual companies. The described project brings together a vertically integrated collaborative team consisting of the end user (U.S. Steel), material producers/suppliers (Allied Mineral Products and Reno Refractories), raw material suppliers (American Metallurgical Services, Minerals Manufacturing), and research organizations (Oak Ridge National Laboratory, National Energy Technology Laboratory and University of Alabama-Birmingham) with the objective of designing and producing new refractory materials based on novel aggregates, improved particle packing, and engineered surface texture. In addition, the recycling and use of spent refractory materials will be investigated and a novel installation technique will be developed taking advantage of new additive manufacturing technology and existing refractory shotcrete technology. It is hoped that the combination of these new technologies will allow for the improvement of the energy, environmental, and economic efficiency of the steel industry while also reducing the environmental footprint of the refractory and steel industries. Additionally, the developed technology is expected to be applicable to other energy intensive industries such as cement, glass, pulp and paper, and non-ferrous metals processing.

Hemrick, James↗

Gate-tunable Veselago interference in a bipolar graphene microcavity

The relativistic charge carriers in monolayer graphene can be manipulated in manners akin to conventional optics. Klein tunneling and Veselago lensing have been previously demonstrated in ballistic graphene pn-junction devices, but collimation and focusing efficiency remains relatively low, preventing realization of advanced quantum devices and controlled quantum interference. Here, we present a graphene microcavity defined by carefully-engineered local strain and electrostatic fields. Electrons are manipulated to form an interference path inside the cavity at zero magnetic field via consecutive Veselago refractions. The observation of unique Veselago interference peaks via transport measurement and their magnetic field dependence agrees with the theoretical expectation. We further utilize Veselago interference to demonstrate localization of uncollimated electrons and thus improvement in collimation efficiency. Our work sheds new light on relativistic single-particle physics and provide a new device concept toward next-generation quantum devices based on manipulation of ballistic electron trajectory.

36 MATERIALS SCIENCE↗

Data for Resourceful and Economical Designing of Fermentation Medium for Lab and Commercial Strains of Yeast from Alternative Feedstock: Transgenic Oilcane

Sugarcane plant engineered to accumulate lipids in its vegetative tissue is being developed as a new bioenergy crop. The new crop would be a source of juice, oil, and cellulosic sugars. However, limited tolerance of industrially recognized yeasts towards inhibitors generated during the processing of lignocellulosic biomass to produce fermentable sugars is a major challenge in developing scalable processes for second-generation drop-in fuel production. To this end, hydrolysates generated from engineered sugarcane—‘oilcane’ bagasse contain added phenolics and fatty acids that further restrict the growth of fermenting microorganisms and necessitate nutrient supplementation and/or detoxification of hydrolysate which makes the fermentation process expensive. Herein, we propose a resourceful and economical approach for growing lab and commercial strains of S. cerevisiae on unrefined cellulosic sugars aerobically and fermentatively. An equal ratio of hydrolysate and juice was found optimum for growth and fermentation by lab and commercial strains of Saccharomyces cerevisiae engineered for xylose fermentation. The industrial strain grew and fermented efficiently under low aeration conditions having an ethanol titer, yield, specific and volumetric productivities of 46.96 ± 0.19 g/l, 0.51 ± 0.00 g/g, 0.27 ± 0.02 g/g.h and 1.95 ± 0.01 g/l.h, respectively, while the lab strain grew better under higher aeration conditions having the ethanol titer, yield, specific and volumetric productivities of 24.93 ± 0.09, 0.27 ± 0.00 g/g, 0.17 ± 0.00 g/g.h and 1.04 ± 0.00 g/l.h, respectively. Acclimation of cultures in a blended medium significantly improved the performance of the yeast strains. The addition of transgenic oilcane juice, which is inedible and rich in amino acids, to the hydrolysate averted the need for expensive nutrient supplementation and detoxification steps of hydrolysate. The approach provides an economical solution to reduce the cost of fermentation at an industrial scale for second-generation drop-in fuel production.

Biomass Analytics↗