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Focused electron beam induced etching (FEBIE) of Nb films produced a maximum etching yield of 3 Nb atoms/e − while varying beam energy, beam current, dwell time, and XeF 2 precursor working pressure. An optimized etch resolution of 17 nm was achieved.
Rhodococcus opacus is a non-model bacterium that is well suited for valorizing lignin. Despite recent advances in our systems-level understanding of its versatile metabolism, studies of its gene functions at a single gene level are still lagging. Elucidating gene functions in non-model organisms is challenging due to limited genetic engineering tools that are convenient to use. To address this issue, we developed a simple gene repression system based on CRISPR interference (CRISPRi). This gene repression system uses a T 7 RNA polymerase system to express a small guide RNA, demonstrating improved repression compared to the previously demonstrated CRISPRi system (i.e., the maximum repression efficiency improved from 58% to 85%). Additionally, our cloning strategy allows for building multiple CRISPRi plasmids in parallel without any PCR step, facilitating the engineering of this GC-rich organism. Using the improved CRISPRi system, we confirmed the annotated roles of four metabolic pathway genes, which had been identified by our previous transcriptomic analysis to be related to the consumption of benzoate, vanillate, catechol, and acetate. Furthermore, we showed our tool’s utility by demonstrating the inducible accumulation of muconate that is a precursor of adipic acid, an important monomer for nylon production. While the maximum muconate yield obtained using our tool was 30% of the yield obtained using gene knockout, our tool showed its inducibility and partial repressibility. In conclusion, our CRISPRi tool will be useful to facilitate functional studies of this non-model organism and engineer this promising microbial chassis for lignin valorization.
Flame-spray-pyrolysis (FSP) is a robust and scalable process to synthesize particles at the commodity-scale. FSP has been used to produce the precursor powders which were converted to the layered structure $(R\bar{3m} \text { phase})$ by a postannealing step in making nickel-rich cathode materials (NCMs). Theoretically, the high flame temperature (normally >1500 K) in FSP can provide adequate energy for the phase conversion from rock-salt to layered structures and potentially enables one-step synthesis. However, the high flame temperature is a critical issue to cause lithium loss and structural degradation, preventing the formation of the layered phase. Here, guided by the gaseous nucleation theory, we implemented several FSP processes with different solution recipes. The layered phase concentration in the as-burned products can be increased with the solution enthalpies. By adding a rapid quench step to suppress the lithium loss and phase degradation, the layered phase can be further increased. This work contributes new ideas to innovating process regarding the process efficiency and throughput of manufacturing cathode materials at a large scale.
Improving the activity and durability of carbon-supported platinum catalysts for the oxygen-reduction reaction (ORR) in acidic electrolytes is crucial to reducing the high overpotentials and power loss over time in proton-exchange membrane fuel cells (PEMFCs). We found that platinum nanoparticle catalyst deposited on an engineered carbon support in the presence of zirconium enabled higher ORR activity and 25% better retention in electrochemically active surface area (ECSA), thereby improving durability. The use of zirconium precursor in the carbon synthesis process led to the formation of atomically dispersed Zr and ZrO 2 nanoparticles on the support. Upon Pt deposition and subsequent heat treatment, the ZrO 2 particles preferentially rearranged on and around the platinum nanoparticles in a chemically reduced form as zirconium suboxide (ZrO 2-x ) surface-decorated nanoclusters, which mitigated Pt nanoparticle coarsening. Finally, analysis of the here-to-fore unknown catalyst structure as well as its performance and durability in a PEM fuel cell membrane electrode assembly (MEA) is discussed.
Abstract Microbial production of esters has recently garnered wide attention, but the current production metrics are low. Evidently, the ester precursors (organic acids and alcohols) can be accumulated at higher titers by microbes like Escherichia coli . Hence, we hypothesized that their ‘direct esterification’ using esterases will be efficient. We engineered esterases from various microorganisms into E. coli , along with overexpression of ethanol and lactate pathway genes. High cell density fermentation exhibited the strains possessing esterase-A (SSL76) and carbohydrate esterase (SSL74) as the potent candidates. Fed-batch fermentation at pH 7 resulted in 80 mg/L of ethyl acetate and 10 mg/L of ethyl lactate accumulation by SSL76. At pH 6, the total ester titer improved by 2.5-fold, with SSL76 producing 225 mg/L of ethyl acetate, and 18.2 mg/L of ethyl lactate, the highest reported titer in E. coli . To our knowledge, this is the first successful demonstration of short-chain ester production by engineering ‘esterases’ in E. coli .
Nuclear Safety is a journal that covers significant issues in the field of nuclear safety. Its primary scope is safety in the design, construction, operation, and decommissioning of nuclear power reactors worldwide and the research and analysis activities that promote this goal, but it also encompasses the safety aspects of the entire nuclear fuel cycle, including fuel fabrication, spent-fuel processing and handling, and nuclear waste disposal, the handling of fissionable materials and radioisotopes, and the environmental effects of all these activities. Table of Contents for this issue follows. GENERAL SAFETY CONSIDERATIONS: 1 A Decision Support System for Maintenance Management of a Boiling-Water Reactor Power Plant, J. H. Shen, A. Ray, and S. Levine; ACCIDENT ANALYSIS: 12 On Prediction of the Ignition Potential of Uranium Metal and Hydride, M. Epstein, W. Luangdilok, M. G. Plys, and H. K. Fauske; 26 An Overview of the Primary Parameters and Methods for Determining Condensation Heat Transfer to Containment Structures, J. Green and K. Almenas; DESIGN FEATURES: 49 Modem Tornado Design of Nuclear and Other Potentially Hazardous Facilities, J. D. Stevenson and Y. Zaho; U.S. NUCLEAR REGULATORY COMMISSION INFORMATION AND ANALYSES: 73 1994 Accident Sequence Precursor Program Results, R. J. Belles, J. W. Cletcher, D. A. Coplnger, B. W. Dolan, J. W. Minarick, and P. D. O'Reilly; ANNOUNCEMENTS: 93 American Institute of Chemical Engineers (AlChE) Spring 1997 National Meeting; 94 European Safety and Reliability Association International Conference on Safety and Reliability ESREL ’97; 95 Criticality Safety Challenges in the Next Decade; 96 21st International Symposium on the Scientific Basis for Nuclear Waste Management; 84 The Authors; 88 Letter to the Editor; 90 Indexes to Nuclear Safety, Volume 36.
The promise of microgravity combustion research is introduced by way of a brief survey of results, the available set of reduced gravity facilities, and plans for experimental capabilities in the Space Station era. The study of fundamental combustion processes in a microgravity environment is a relatively new scientific endeavor. A few simple, precursor experiments were conducted in the early 1970's. Today the advent of the U.S. space shuttle and the anticipation of the Space Station Freedom provide for scientists and engineers a special opportunity, in the form of long duration microgravity laboratories, and need, in the form of spacecraft fire safety and a variety of terrestrial applications, to pursue fresh insight into the basic physics of combustion. The microgravity environment enables a new range of experiments to be performed since buoyancy-induced flows are nearly eliminated, normally obscured forces and flows may be isolated, gravitational settling or sedimentation is nearly eliminated, and larger time or length scales in experiments become permissible. The range of experiments completed to date was not broad, but is growing. Unexpected phenomena have been observed often in microgravity combustion experiments, raising questions about the degree of accuracy and completion of our classical understanding and our ability to estimate spacecraft fire hazards. Because of the field's relative immaturity, instrumentation has been restricted primarily to high-speed photography. To better explain these findings, more sophisticated diagnostic instrumentation, similar to that evolving in terrestrial laboratories, is being developed for use on Space Station Freedom and, along the way, in existing microgravity facilities.
The study of fundamental combustion processes in a microgravity environment is a relatively new scientific endeavor. A few simple, precursor experiments were conducted in the early 1970's. Today the advent of the U.S. space shuttle and the anticipation of the Space Station Freedom provide for scientists and engineers a special opportunity -- in the form of long duration microgravity laboratories -- and need -- in the form of spacecraft fire safety and a variety of terrestrial applications -- to pursue fresh insight into the basic physics of combustion. Through microgravity, a new range of experiments can be performed since: (1) Buoyancy-induced flows are nearly eliminated; (2) Normally obscured forces and flows may be isolated; (3) Gravitational settling or sedimentation is nearly eliminated; and (4) Larger time or length scales in experiments become permissible.
Soot formation is a complex dynamic and intermittent process determined by properties of the fuel, combustor design, and combustor operation. Although the major steps in soot formation (i.e., formation of precursors, inception, growth and evolution) are similar for a variety of carbonaceous fuels, applications, and operating conditions, it remains unclear when the temporal transition between these steps occurs. An engineering prediction tool coupled with computational fluid physics (CFD), therefore needs to accurately model all these complex steps. To develop such a model, we propose the time-history concept for understanding the time dependency of soot formation as a function of local properties (i.e., temperature, velocity, local fuel air ratio, etc.). We continue our previous work with modeling the DLR aero-combustor [1] with our updated in-house CFD code, Open National Combustion Code (OpenNCC), that now includes a Multiple Time-Scale Flamelet Progress Variable approach and a the semi-empirical two-equation soot model. We injected massless tracer particles upstream of the injector region of the combustor to collect time-history statistics of the solution variables. The correlations between the collected statistics with respect to the experimental soot volume fraction data showed that time-history effect of certain flow variables, including turbulent kinetic energy (TKE), and multiple species is indeed important for soot formation. We then conducted a time-history based correlation analysis to determine the key species and the concentration ranges critical for soot formation (C6H5-based nucleation, acetylene-based surface growth, and oxidation with OH and O2). Based on the time-history correlation coefficient (THCC) analysis, we propose possible modifications to improve the current two-equation model.
Nuclear Safety is a journal that covers significant issues in the field of nuclear safety. Its primary scope is safety in the design, construction, operation, and decommissioning of nuclear power reactors worldwide and the research and analysis activities that promote this goal, but it also encompasses the safety aspects of the entire nuclear fuel cycle, including fuel fabrication, spent-fuel processing and handling, and nuclear waste disposal, the handling of fissionable materials and radioisotopes, and the environmental effects of all these activities. Table of Contents for this issue follows. GENERAL SAFETY CONSIDERATIONS: 1 A Decision Support System for Maintenance Management of a Boiling-Water Reactor Power Plant, J. H. Shen, A. Ray, and S. Levine; ACCIDENT ANALYSIS: 12 On Prediction of the Ignition Potential of Uranium Metal and Hydride, M. Epstein, W. Luangdilok, M. G. Plys, and H. K. Fauske; 26 An Overview of the Primary Parameters and Methods for Determining Condensation Heat Transfer to Containment Structures, J. Green and K. Almenas; DESIGN FEATURES: 49 Modem Tornado Design of Nuclear and Other Potentially Hazardous Facilities, J. D. Stevenson and Y. Zaho; U.S. NUCLEAR REGULATORY COMMISSION INFORMATION AND ANALYSES: 73 1994 Accident Sequence Precursor Program Results, R. J. Belles, J. W. Cletcher, D. A. Copinger, B. W. Dolan, J. W. Minarick, and P. D. O'Reilly; ANNOUNCEMENTS: 93 American Institute of Chemical Engineers (AlChE) Spring 1997 National Meeting; 94 European Safety and Reliability Association International Conference on Safety and Reliability ESREL ’97; 95 Criticality Safety Challenges in the Next Decade; 96 21st International Symposium on the Scientific Basis for Nuclear Waste Management; 84 The Authors; 88 Letter to the Editor; 90 Indexes to Nuclear Safety, Volume 36.
Efficient conversion of inexpensive feedstocks to valuable chemicals by microbes is critical for a robust bioeconomy, but the ability to rationally design bacteria is hampered by insufficient knowledge of how post translational modifications (PTMs) control bacterial protein function and thus bioproduction phenotypes. Our study will focus on the lysine acetylation, a ubiquitous bacterial PTM that can affect the function of enzymes in central metabolism that are often critical for bioproduction processes, disrupt transcriptional regulation, and reduce translation. However, most lysine acetylation data is observational, which means that we do not know when, how, and what specific acetylated residues affect protein function and bacterial physiology. For our model host, we will use a Pseudomonas putida strain that we previously engineered to convert lignocellulosic feedstocks into chemicals such as itaconic acid (ITA). With this strain, we use a dynamic two-stage bioproduction process in which ITA is produced during a non-growth associated production phase. Production is highest during growth stages when lysine acetylation is low in other organisms (early stationary phase) and stalls in conditions where acetylation is highest (late stationary phase). The switch from high to stalled ITA production is also correlated with an unexpected increase in acetate levels – the precursor to non-enzymatic lysine acetylation. As such, we predict that lysine acetylation plays a substantial role in regulating the metabolic pathways required for ITA production. We will develop a generalizable approach that combines high-throughput genetic screens and cutting-edge genome engineering with state-of-the-art proteomics, metabolomics, and genetic code expansion methods to identify and modulate lysine acetylation patterns in bacteria. Ultimately, these strategies aim to manipulate protein expression and acetylation patterns to enhance bioproduction phenotypes (e.g., sustained ITA production in late stationary phase).
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Cost-effective production of the highly effective anti-cancer drug, paclitaxel (Taxol ® ), remains limited despite growing global demands. Low yields of the critical taxadiene precursor remains a key bottleneck in microbial production. In this study, the key challenge of poor taxadiene synthase (TASY) solubility in S. cerevisiae was revealed, and the strains were strategically engineered to relieve this bottleneck. Multi-copy chromosomal integration of TASY harbouring a selection of fusion solubility tags improved taxadiene titres 22-fold, up to 57 ± 3 mg/L at 30 °C at microscale, compared to expressing a single episomal copy of TASY. The scalability of the process was highlighted through achieving similar titres during scale up to 25 mL and 250 mL in shake flask and bioreactor cultivations, respectively at 20 and 30 °C. Maximum taxadiene titres of 129 ± 15 mg/L and 127 mg/L were achieved through shake flask and bioreactor cultivations, respectively, of the optimal strain at a reduced temperature of 20 °C. The results of this study highlight the benefit of employing a combination of molecular biology and bioprocess tools during synthetic pathway development, with which TASY activity was successfully improved by 6.5-fold compared to the highest literature titre in S. cerevisiae cell factories.
The primary objective of the Center for Modelling of Turbulence and Transition (CMOTT) is to further the understanding of turbulence theory for engineering applications. One important foundation is the establishment of a data base encompassing the multitude of existing models as well as newly proposed ideas. The research effort described is a precursor to an extended survey of two equation turbulence models in the presence of a separated shear layer. Recently, several authors have examined the performance of two equation models in the context of the backward facing step flow. Conflicting results, however, demand that further attention is necessary to properly understand the behavior and limitations of this popular technique, especially the low Reynolds number formulations. The objective is to validate an incompressible Navier Stokes code for use as a numerical test-bed. In turn, this code will be used for analyzing the performance of several two equation models.
The development of thin film solar cells on flexible, lightweight, space-qualified substrates provides an attractive cost solution to fabricating solar arrays with high specific power, (W/kg). The use of a polycrystalline chalcopyrite absorber layer for thin film solar cells is considered as the next generation photovoltaic devices. At NASA GRC we have focused on the development of new single source precursors (SSP) and their utility to deposit the chalcopyrite semi-conducting layer (CIS) onto flexible substrates for solar cell fabrication. The syntheses and thermal modulation of SSPs via molecular engineering is described. Thin-film fabrication studies demonstrate the SSPs can be used in a spray CVD (chemical vapor deposition) process, for depositing CIS at reduced temperatures, which display good electrical properties, suitable for PV (photovoltaic) devices.
The development of thin-film solar cells on flexible, lightweight, space-qualified substrates provides an attractive cost solution to fabricating solar arrays with high specific power, (W/kg). The use of a polycrystalline chalcopyrite absorber layer for thin film solar cells is considered as the next generation photovoltaic devices. A key technical issues outlined in the 2001 U.S. Photovoltaic Roadmap, is the need to develop low cost, high throughput manufacturing for high-efficiency thin film solar cells. At NASA GRC we have focused on the development of new single-source-precursors (SSPs) and their utility to deposit the chalcopyrite semi-conducting layer (CIS) onto flexible substrates for solar cell fabrication. The syntheses and thermal modulation of SSPs via molecular engineering is described. Thin-film fabrication studies demonstrate the SSPs can be used in a spray CVD process, for depositing CIS at reduced temperatures, which display good electrical properties, suitable for PV devices.
The nuclear thermal rocket (NTR) represents the next "evolutionary step" in high performance rocket propulsion. Unlike conventional chemical rockets that produce their energy through combustion, the NTR derives its energy from fission of Uranium-235 atoms contained within fuel elements that comprise the engine s reactor core. Using an "expander" cycle for turbopump drive power, hydrogen propellant is raised to a high pressure and pumped through coolant channels in the fuel elements where it is superheated then expanded out a supersonic nozzle to generate high thrust. By using hydrogen for both the reactor coolant and propellant, the NTR can achieve specific impulse (Isp) values of ~900 seconds (s) or more - twice that of today s best chemical rockets. From 1955 - 1972, twenty rocket reactors were designed, built and ground tested in the Rover and NERVA (Nuclear Engine for Rocket Vehicle Applications) programs. These programs demonstrated: (1) high temperature carbide-based nuclear fuels; (2) a wide range of thrust levels; (3) sustained engine operation; (4) accumulated lifetime at full power; and (5) restart capability - all the requirements needed for a human Mars mission. Ceramic metal "cermet" fuel was pursued as well, as a backup option. The NTR also has significant "evolution and growth" capability. Configured as a "bimodal" system, it can generate its own electrical power to support spacecraft operational needs. Adding an oxygen "afterburner" nozzle introduces a variable thrust and Isp capability and allows bipropellant operation. In NASA s recent Mars Design Reference Architecture (DRA) 5.0 study, the NTR was selected as the preferred propulsion option because of its proven technology, higher performance, lower launch mass, versatile vehicle design, simple assembly, and growth potential. In contrast to other advanced propulsion options, no large technology scale-ups are required for NTP either. In fact, the smallest engine tested during the Rover program - the 25,000 lbf (25 klbf) "Pewee" engine is sufficient when used in a clustered engine arrangement. The "Copernicus" crewed spacecraft design developed in DRA 5.0 has significant capability and a human exploration strategy is outlined here that uses Copernicus and its key components for precursor near Earth object (NEO) and Mars orbital missions prior to a Mars landing mission. The paper also discusses NASA s current activities and future plans for NTP development that include system-level Technology Demonstrations - specifically ground testing a small, scalable NTR by 2020, with a flight test shortly thereafter.
The NTR represents the next evolutionary step in high performance rocket propulsion. It generates high thrust and has a specific impulse (Isp) of approx.900 seconds (s) or more V twice that of today s best chemical rockets. The technology is also proven. During the previous Rover and NERVA (Nuclear Engine for Rocket Vehicle Applications) nuclear rocket programs, 20 rocket reactors were designed, built and ground tested. These tests demonstrated: (1) a wide range of thrust; (2) high temperature carbide-based nuclear fuel; (3) sustained engine operation; (4) accumulated lifetime; and (5) restart capability V all the requirements needed for a human mission to Mars. Ceramic metal cermet fuel was also pursued, as a backup option. The NTR also has significant growth and evolution potential. Configured as a bimodal system, it can generate electrical power for the spacecraft. Adding an oxygen afterburner nozzle introduces a variable thrust and Isp capability and allows bipropellant operation. In NASA s recent Mars Design Reference Architecture (DRA) 5.0 study, the NTR was selected as the preferred propulsion option because of its proven technology, higher performance, lower launch mass, simple assembly and mission operations. In contrast to other advanced propulsion options, NTP requires no large technology scale-ups. In fact, the smallest engine tested during the Rover program V the 25,000 lbf (25 klbf) Pewee engine is sufficient for human Mars missions when used in a clustered engine arrangement. The Copernicus crewed spacecraft design developed in DRA 5.0 has significant capability and a human exploration strategy is outlined here that uses Copernicus and its key components for precursor near Earth asteroid (NEA) and Mars orbital missions prior to a Mars landing mission. Initially, the basic Copernicus vehicle can enable reusable 1-year round trip human missions to candidate NEAs like 1991 JW and Apophis in the late 2020 s to check out vehicle systems. Afterwards, the Copernicus spacecraft and its 2 key components, now configured as an Earth Return Vehicle / propellant tanker, would be used for a short round trip (approx.18 - 20 months)/short orbital stay (60 days) Mars / Phobos survey mission in 2033 using a split mission approach. The paper also discusses NASA s current Foundational Technology Development activities and its pre-decisional plans for future system-level Technology Demonstrations that include ground testing a small (approx.7.5 klbf) scalable NTR before the decade is out with a flight test shortly thereafter.