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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 19 records

Development of a shuttle plasmid without host restriction sites for efficient transformation and heterologous gene expression in Clostridium cellulovorans

Clostridium cellulovorans capable of producing large amounts of acetate and butyrate from cellulose is a promising candidate for biofuels and biochemicals production from lignocellulosic biomass. However, the restriction modification (RM) systems of C. cellulovorans hindered the application of existing shuttle plasmids for metabolic engineering of this organism. To overcome the hurdle of plasmid digestion by host, a new shuttle plasmid (pYL001) was developed to remove all restriction sites of two major RM systems of C. cellulovorans , Cce 743 I and Cce 743 II. The pYL001 plasmid remained intact after challenge by C. cellulovorans cell extract. Post-electroporation treatments and culturing conditions were also modified to improve cell growth and colony formation on agar plates. With the improvements, the pYL001 plasmid, without in-vivo methylation, was readily transformed into C. cellulovorans with colonies of recombinant cells formed on agar plates within 24 h. Three pYL001-derived recombinant plasmids free of Cce 743 I/ Cce 743 II restriction sites, after synonymous mutation of the heterologous genes, were constructed and transformed into C. cellulovorans . Functional expression of these genes was confirmed with butanol and ethanol production from glucose in batch fermentations by the transformants. In conclusion, the pYL001 plasmid and improved transformation method can facilitate further metabolic engineering of C. cellulovorans for cellulosic butanol production.

09 BIOMASS FUELS↗

Development of exosome surface display technology in living human cells

Surface display technology is an emerging key player in presenting functional proteins for targeted drug delivery and therapy. Although a number of technologies exist, a desirable mammalian surface display system is lacking. Exosomes are extracellular vesicles that facilitate cell–cell communication and can be engineered as nano-shuttles for cell-specific delivery. In this study, we report the development of a novel exosome surface display technology by exploiting mammalian cell secreted nano-vesicles and their trans-membrane protein tetraspanins. By constructing a set of fluorescent reporters for both the inner and outer surface display on exosomes at two selected sites of tetraspanins, we demonstrated the successful exosomal display via gene transfection and monitoring fluorescence in vivo. We subsequently validated our system by demonstrating the expected intracellular partitioning of reporter protein into sub-cellular compartments and secretion of exosomes from human HEK293 cells. Lastly, we established the stable engineered cells to harness the ability of this robust system for continuous production, secretion, and uptake of displayed exosomes with minimal impact on human cell biology. In sum, our work paved the way for potential applications of exosome, including exosome tracking and imaging, targeted drug delivery, as well as exosome-mediated vaccine and therapy.

60 APPLIED LIFE SCIENCES↗

High temperature thick film sensor development based on doped lanthanum chromites refractory semiconductors materials

High temperature advanced sensing materials have generated high demand due the high accuracy temperature measurements requirements for process optimization, controlling and sensing. Some technological applications of harsh conditions sensing include monitoring tiles of space shuttles, rotating bearings in aircraft engines, turbines, jet engines dynamics and chemical reactors. High temperature conditions limit the sensing strategies, where typically traditional metal thermocouples are unstable, and the sensing options are limited to optical spectroscopy methods. Recently, refractory semiconductors thick- and thin-film thermocouples have been developed and, in many cases, preferred over conventional metallic thermocouples due their spatial resolution, and capability of direct deposition on any surface. Rare earth chromites ceramics materials, exhibit some properties of interest for high temperature sensing technologies development, such as: high microstructure and sintering stability, excellent conductive behavior at high temperatures, and matching thermal expansion coefficients relative to other conductors and refractory ceramics. In this work, high performance ultra-high temperature thermocouples using p-type and n-type doped lanthanum chromites materials were fabricated and tested at temperatures up to 1500 o C. Thermoelectric voltage, Seebeck Coefficients were established for all devices, evidencing high stability and performance in prolongated operational time and harsh conditions.

20 FOSSIL-FUELED POWER PLANTS↗

Facet-dependent electron transfer induces distinct arsenic reallocations on hematite

The interfacial electron transfer (ET) between electron shuttling compounds and iron (Fe) oxyhydroxides plays a crucial role in the reductive dissolution of Fe minerals and the fate of surface-bound arsenic (As). However, the impact of exposed facets of highly crystalline hematite on reductive dissolution and As immobilization is poorly understood. In this study, we systematically investigated the interfacial processes of the electron shuttling compound cysteine (Cys) on various facets of hematite and the reallocations of surface-bound As(III) or As(V) on the respective surfaces. Our results demonstrate that the ET process between Cys and hematite generates Fe(II) and leads to reductive dissolution, with more Fe(II) generated on {001} facets of exposed hematite nanoplates (HNPs). Reductive dissolution of hematite leads to significantly enhanced As(V) reallocations on hematite. Nevertheless, upon the addition of Cys, a raipd release of As(III) can be halted by its prompt re-adsorption, leaving the extent of As(III) immobilization on hematite unchanged throughout the course of reductive dissolution. This is due to that Fe(II) can form new precipitates with As(V), a process that is facet-dependent and influenced by water chemistry. Electrochemical analysis reveals that HNPs exhibit higher conductivity and ET ability, which is beneficial for reductive dissolution and As reallocations on hematite. In conclusion, these findings highlight the facet-dependent reallocations of As(III) and As(V) facilitated by electron shuttling compounds and have implications for the biogeochemical processes of As in soil and subsurface environments.

54 ENVIRONMENTAL SCIENCES↗

30-kW Maintenance-Free Stirling Engine for High-Performance Dish Concentrating Solar

This final report summarizes the final results of the 30-kW Maintenance-Free Stirling Engine for High-Performance Dish Concentrating Solar project performed by Infinia Corporation for the U.S. Department of Energy under contract DE-FC36-08GO18032 during the project period of March 1, 2008 – March 31, 2012. Over the course of the project, Infinia Corporation achieved substantial progress in the design, fabrication, assembly, testing and evaluation of a 30-kW multi-cylinder, free-piston Stirling engine (MCFPSE) prototype. In this effort, prior MCFPSE conceptual design approaches were adapted for the development of a 30-kW design, thereby producing a cohesive design package and detailed performance specifications. Numerical analysis using the Stirling Engine Analysis and Optimization Code (SAGE) was completed culminating in the selection of a fully balanced 6-cylinder configuration composed of two 3-cylinder engine modules. The model was refined throughout the program to include all wall conduction losses, piston and rod seal losses, appendix, shuttle and pumping losses, and piston radiation loss to closely represent the real system. Performance mapping of the engine was then completed using the detailed model. Based on knowledge and experience from its existing commercial products, and using the results from the extensive numerical modeling discussed above, Infinia successfully fabricated, assembled and tested two sets of 30-kW engine modules. One of these configurations was driven by an external power supply and operated as a cooler in order to verify the mechanical integrity of the system. In the engine configuration, induction heaters provided the energy to the heater head to operate the engine. Considerable effort was expended on correcting problems associated with contact and rubbing of internal components, identification and subsequent elimination of internal leaks, and undesirable amounts of piston bias. Many details concerning the fabrication and testing efforts are considered proprietary. As a result, those findings are not disclosed herein but rather can be found in proprietary reports supplied to the DOE on a periodic basis. The prototype did not fully demonstrate performance nor did it realize the projected cost targets. Furthermore, the basis of the mechanical and electrical architecture of the 30-kW multi-cylinder prototype design is now at least two generations old. Alternatively, Infinia’s 8-kW Bonneville generator, which was successfully demonstrated in April 2012, and is slated to enter commercial production in late 2013, offers immediate benefits of lower cost and significantly higher power density such that the generator will likely meet DOE’s Sunshot LCOE target goal of $\$$0.07-0.10/kWh by 2015 and $\$$0.05-0.07/kWh by 2020. Therefore, the design of a 30-kW Stirling engine as disclosed in the project will not be pursued further. However, Infinia remains keenly interested in developing higher power Stirling machines in the future based on the new Bonneville mechanical design.

14 SOLAR ENERGY↗

Electrolytes with Solvating Inner Sheath Engineering for Practical Na–S Batteries

Sodium–sulfur (Na–S) batteries with durable Na-metal stability, shuttle-free cyclability, and long lifespan are promising to large-scale energy storages. However, meeting these stringent requirements poses huge challenges with the existing electrolytes. Herein, a localized saturated electrolyte (LSE) is proposed with 2-methyltetrahydrofuran (MeTHF) as an inner sheath solvent, which represents a new category of electrolyte for Na–S system. Unlike the traditional high concentration electrolytes, the LSE is realized with a low salt-to-solvent ratio and low diluent-to-solvent ratio, which pushes the limit of localized high concentration electrolyte (LHCE). The appropriate molecular structure and solvation ability of MeTHF regulate a saturated inner sheath, which features a reinforced coordination of Na + to anions, enlarged Na + -solvent distance, and weakened anion-diluent interaction. Such electrolyte configuration is found to be the key to build a sustainable interphase and a quasi-solid–solid sulfur redox process, making a dendrite-inhibited and shuttle-free Na–S battery possible. With this electrolyte, pouch cells with decent cycling performance under rather demanding conditions are demonstrated.

25 ENERGY STORAGE↗

Nontraditional Approaches To Enable High-Energy and Long-Life Lithium–Sulfur Batteries

In this study, lithium-sulfur (Li-S) batteries are promising for automotive applications due to their high theoretical energy density (2600 Wh/kg). In addition, the natural abundance of sulfur could mitigate the global raw material supply chain challenge of commercial lithium-ion batteries that use critical elements, such as nickel and cobalt. However, due to persistent polysulfide shuttling and uncontrolled lithium dendrite growth, Li-S batteries using nonencapsulated sulfur cathodes and conventional ether-based electrolytes suffer from rapid cell degradation upon cycling. Despite significant improvements in recent decades, there is still a big gap between lab research and commercialization of the technology. To date, the reported cell energy densities and cycling life of practical Li-S pouch cells remain largely unsatisfactory. Traditional approaches to improving Li-S performance are primarily focused on confining polysulfides using electronically conductive hosts. However, these micro- and mesoporous hosts suffer from limited pore volume to accommodate high sulfur loading and the associated volume change during cycling. Moreover, they fail to balance adsorption-conversion of polysulfides during charge-discharge, leading to the formation of massive dead sulfur. Such hosts are themselves electrochemically inactive, which decreases the practical energy density. In contrast, a series of nontraditional approaches, paired with advances in multiscale mechanistic understanding, have recently demonstrated exciting performance outcomes not only in conventional coin cells but also in practical pouch cells. In this Account, we first introduce our novel cathode design strategies to overcome polysulfide shuttling and sluggish redox kinetics in thick S cathodes via selenium-sulfur chemistry and cathode host engineering. Next, we gain a mechanistic understanding of Li-S batteries in various types of electrolytes via a series of spectroscopic, nuclear magnetic resonance, and electrochemical methods. Meanwhile, a novel cathode solid electrolyte interphase encapsulation strategy via nonviscous highly fluorinated ether-based electrolyte is introduced. The established selection rule by investigating how solvating power retards the shuttle effect and induces robust cathode/solid-electrolyte interphase formation is also included. We then discuss how the synergistic interactions between rational cathode structures and electrolytes can be exploited to tailor the reaction pathways and kinetics of S cathodes under high mass loading and lean electrolyte conditions. In addition, a novel interlayer design to simultaneously overcome degradation processes (polysulfide shuttling and lithium dendrite formation) and accelerate redox reaction kinetics is presented. Finally, this Account concludes with an overview of the challenges and strategies to develop Li-S pouch cells with high practical energy density, long cycle life, and fast-charging capability.

25 ENERGY STORAGE↗

Clean Cities Guide to Alternative Fuel and Advanced Medium- and Heavy-Duty Vehicles

Today's fleets are increasingly interested in medium-duty and heavy-duty vehicles that use alternative fuels or advanced technologies that can help reduce operating costs, meet emissions requirements, improve fleet sustainability, and support U.S. energy independence. Vehicle and engine manufacturers are responding to this interest with a wide range of options across a steadily growing number of vehicle applications. This guide provides an overview of alternative fuel power systems--including engines, microturbines, electric motors, and fuel cells--and hybrid propulsion systems. The guide also offers a list of individual medium- and heavy-duty vehicle models listed by application, along with associated manufacturer contact information, fuel type(s), power source(s), and related information.

All-Electric↗

A Molecular Biology Tool Kit for the Phototrophic Firmicute Heliobacterium modesticaldum

The heliobacteria are members of the bacterial order Clostridiales and form the only group of phototrophs in the phylum Firmicutes . Several physiological and metabolic characteristics make them an interesting subject of investigation, including their minimalist photosynthetic system, nitrogen fixation abilities, and ability to reduce toxic metals. Although the species Heliobacterium modesticaldum is an excellent candidate as a model system for the family Heliobacteriaceae , since an annotated genome and transcriptomes are available, studies in this organism have been hampered by the lack of genetic tools. We adapted techniques for genetic manipulation of related clostridial species for use with H. modesticaldum . Five heliobacterial DNA methyltransferase genes were expressed in an Escherichia coli strain engineered as a conjugative plasmid donor for broad-host-range plasmids. Premethylation of the shuttle vectors before conjugation into H. modesticaldum is absolutely required for production of transconjugant colonies. The introduced shuttle vectors are maintained stably and can be recovered using a modified minipreparation procedure developed to inhibit endogenous DNase activity. Moreover, we describe the formulation of various growth media, including a defined medium for metabolic studies and isolation of auxotrophic mutants. Heliobacteria are anoxygenic phototrophic bacteria with the simplest known photosynthetic apparatus. They are unique in using bacteriochlorophyll g as their main pigment and lacking a peripheral antenna system. Until now, research on this organism has been hampered by the lack of a genetic transformation system. Without such a system, gene knockouts, site-directed mutations, and gene expression studies cannot be performed to help us further understand or manipulate the organism. Here we report the genetic transformation of a heliobacterium, which should enable future genetic studies in this unique phototrophic organism.

59 BASIC BIOLOGICAL SCIENCES↗

Quantification of residual stress from photonic signatures of fused silica

A commercially available grey-field polariscope (GFP) instrument for photoelastic examination is used to assess impact damage inflicted upon the outer-most pane of Space Shuttle windows made from fused silica. A method and apparatus for calibration of the stress-optic coefficient using four-point bending is discussed. The results are validated on known material (acrylic) and are found to agree with literature values to within 6%. The calibration procedure is then applied to fused-silica specimens and the stress-optic coefficient is determined to be 2.43 ± 0.54 × 10{sup −12} Pa{sup −1}. Fused silica specimens containing impacts artificially made at NASA’s Hypervelocity Impact Technology Facility (HIT-F), to simulate damage typical during space flight, are examined. The damage sites are cored from fused silica window carcasses and examined with the GFP. The calibrated GFP measurements of residual stress patterns surrounding the damage sites are presented.

42 ENGINEERING↗

Improving the Transformation Efficiency of Synechococcus sp. PCC 7002 via Methylome-Guided Premethylation of DNA

Cyanobacteria are promising microbial platforms for a diverse set of biotechnology applications, from living materials to photosynthetic chemical production, but are less well characterized than commonly engineered microbes such as Escherichia coli. This study facilitates genetic engineering in Synechococcus sp. PCC 7002, a fast-growing, halotolerant, and naturally competent strain, by identifying ten native methylation motifs and designing shuttle strains that mimic the native methylation state by expressing a subset of heterologous methyltransferases. DNA methylation in E. coli with as few as two active methyltransferases increased transformation efficiency up to 30-fold across four distinct integration sites in PCC 7002. This work provides an experimental framework to bypass native restriction-modification systems for efficient genome editing and metabolic engineering in nonmodel bacteria.

59 BASIC BIOLOGICAL SCIENCES↗

Protein–Protein Complex Stability Controls Substrate Scope in a β‐Ketoacyl‐ACP Reductase Specific for Medium Chains

Assembly-line enzymes carry out multistep synthesis of important metabolites by using acyl carrier proteins (ACPs) to shuttle intermediates along defined sequences of active sites. Despite longstanding interest in reprogramming these systems for metabolic engineering and biosynthetic chemistry, the mechanisms underlying their reaction order remain poorly understood and difficult to control. In this work, we describe a β-ketoacyl-ACP reductase from Pseudomonas putida (PpFabG4) with an unusual selectivity for medium chains and use it to explore the molecular basis of substrate specificity in enzymes that pull intermediates from fatty acid synthesis, a common route to specialized products. X-ray crystallography shows no obvious barriers to short-chain binding. Molecular simulations and supporting mutational analyses indicate that substrate preference arises instead from a weak enzyme–ACP interaction that is stabilized by medium acyl chains but not by short chains. Indeed, mutations that strengthen this interaction for PpFabG4 or weaken it for EcFabG, an Escherichia coli β-ketoacyl-ACP reductase with a broad substrate specificity, can enhance or reduce activity on short-chain substrates by over 100-fold. Our findings show how the stability of enzyme-ACP interactions can control substrate scope in promiscuous enzymes and guide the exchange of intermediates between (and within) assembly-line systems.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Characterization of ferredoxins involved in electron transfer pathways for nitrogen fixation implicates differences in electronic structure in tuning 2[4Fe 4S] Fd activity

Ferredoxins (Fds) are small proteins which shuttle electrons to pathways like biological nitrogen fixation. Physical properties tune the reactivity of Fds with different pathways, but knowledge on how these properties can be manipulated to engineer new electron transfer pathways is lacking. Recently, we showed that an evolved strain of Rhodopseudomonas palustris uses a new electron transfer pathway for nitrogen fixation. This pathway involves a variant of the primary Fd of nitrogen fixation in R. palustris, Fer1, in which threonine at position 11 is substituted for isoleucine (Fer1 T11I ). To understand why this substitution in Fer1 enables more efficient electron transfer, we used in vivo and in vitro methods to characterize Fer1 and Fer1 T11I . Electrochemical characterization revealed both Fer1 and Fer1 T11I have similar redox transitions (–480 mV and – 550 mV), indicating the reduction potential was unaffected despite the proximity of T11 to an iron-sulfur (Fe—S) cluster of Fer1. Additionally, disruption of hydrogen bonding around an Fe—S cluster in Fer1 by substituting threonine with alanine (T11A) or valine (T11V) did not increase nitrogenase activity, indicating that disruption of hydrogen bonding does not explain the difference in activity observed for Fer1 T11I . Electron paramagnetic resonance spectroscopy studies revealed key differences in the electronic structure of Fer1 and Fer1 T11I , which indicate changes to the high spin states and/or spin-spin coupling between the Fe—S clusters of Fer1. Finally, our data implicates these electronic structure differences in facilitating electron flow and sets a foundation for further investigations to understand the connection between these properties and intermolecular electron transfer.

59 BASIC BIOLOGICAL SCIENCES↗

Superconducting Shuttle-Flux Shift Register for Race Logic and Its Applications

Here this paper presents a superconducting, magnetically-coupled, shuttle-flux shift register (SF-SR) that stores single flux quantum (SFQ) pulses. This shift register has a DC bias operating margin of ±34% at 10 GHz, with a power dissipation of 3.6 μW and 38% fewer Josephson junctions (JJs) when scaled up to multiple stages compared to a data flip-flop (DFF) based shift register. The clock input is inductively coupled and is independent from the data input. We then present three applications for our SF-SR. In the first application, we add two non-destructive readout (NDRO) cells to construct a buffer that temporarily stores the temporal information of a series of race logic (RL) pulses. The second application is a pseudo-random number generator based on a linear function shift register (LFSR). The third application is N parallel SF-SRs that can act similar to a deserializer or instead can emulate a single SF-SR of N times higher clock frequency. These three applications motivate deep shift registers with many shifting intervals, which our SF-SR can implement with fewer JJs and lower power consumption compared to DFF-based shift registers.

42 ENGINEERING↗

Mesoscale Battery Science: The Behavior of Electrode Particles Caught on a Multispectral X-ray Camera

Here, functional materials and devices are usually morphologically complex and chemically heterogeneous. Their structures are often designed to be hierarchical because of the desired functionalities, which usually require many different components to work together in a coherent manner. The lithium ion battery, as an energy storage device, is a very typical example of this kind of structure. In a lithium ion battery, the cathode, anode, and separator are soaked in a liquid electrolyte, facilitating the back and forward shuttling of the lithium ions for energy storage and release. The desired performance of a lithium ion battery has many different aspects that need to be engineered and balanced depending on the targeted applications. In most cases, the cathode material has become the limiting factor for further improvements and, thus, has attracted intense attention from the research community.

25 ENERGY STORAGE↗

Distinct Physiological Roles of the Three Ferredoxins Encoded in the Hyperthermophilic Archaeon Thermococcus kodakarensis

Control of electron flux is critical in both natural and bioengineered systems to maximize energy gains. Both small molecules and proteins shuttle high-energy, low-potential electrons liberated during catabolism through diverse metabolic landscapes. Ferredoxin (Fd) proteins—an abundant class of Fe-S-containing small proteins—are essential in many species for energy conservation and ATP production strategies. It remains difficult to model electron flow through complicated metabolisms and in systems in which multiple Fd proteins are present. The overlap of activity and/or limitations of electron flux through each Fd can limit physiology and metabolic engineering strategies. Here we establish the interplay, reactivity, and physiological role(s) of the three ferredoxin proteins in the model hyperthermophile Thermococcus kodakarensis . We demonstrate that the three loci encoding known Fds are subject to distinct regulatory mechanisms and that specific Fds are utilized to shuttle electrons to separate respiratory and energy production complexes during different physiological states. The results obtained argue that unique physiological roles have been established for each Fd and that continued use of T. kodakarensis and related hydrogen-evolving species as bioengineering platforms must account for the distinct Fd partnerships that limit flux to desired electron acceptors. Extrapolating our results more broadly, the retention of multiple Fd isoforms in most species argues that specialized Fd partnerships are likely to influence electron flux throughout biology. High-energy electrons liberated during catabolic processes can be exploited for energy-conserving mechanisms. Maximal energy gains demand these valuable electrons be accurately shuttled from electron donor to appropriate electron acceptor. Proteinaceous electron carriers such as ferredoxins offer opportunities to exploit specific ferredoxin partnerships to ensure that electron flux to critical physiological pathways is aligned with maximal energy gains. Most species encode many ferredoxin isoforms, but very little is known about the role of individual ferredoxins in most systems. Our results detail that ferredoxin isoforms make largely unique and distinct protein interactions in vivo and that flux through one ferredoxin often cannot be recovered by flux through a different ferredoxin isoform. The results obtained more broadly suggest that ferredoxin isoforms throughout biological life have evolved not as generic electron shuttles, but rather serve as selective couriers of valuable low-potential electrons from select electron donors to desirable electron acceptors.

59 BASIC BIOLOGICAL SCIENCES↗

Mapping suitability areas for concentrated solar power plants using remote sensing data

The political push to increase power generation from renewable sources such as solar energy requires knowing the best places to site new solar power plants with respect to the applicable regulatory, operational, engineering, environmental, and socioeconomic criteria. Therefore, in this paper, we present applications of remote sensing data for mapping suitability areas for concentrated solar power plants. Our approach uses digital elevation model derived from NASA s Shuttle Radar Topographic Mission (SRTM) at a resolution of 3 arc second (approx. 90m resolution) for estimating global solar radiation for the study area. Then, we develop a computational model built on a Geographic Information System (GIS) platform that divides the study area into a grid of cells and estimates site suitability value for each cell by computing a list of metrics based on applicable siting requirements using GIS data. The computed metrics include population density, solar energy potential, federal lands, and hazardous facilities. Overall, some 30 GIS data are used to compute eight metrics. The site suitability value for each cell is computed as an algebraic sum of all metrics for the cell with the assumption that all metrics have equal weight. Finally, we color each cell according to its suitability value. Furthermore, we present results for concentrated solar power that drives a stream turbine and parabolic mirror connected to a Stirling Engine.

14 SOLAR ENERGY↗