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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
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

Alpha ketoacid decarboxylases: Diversity, structures, reaction mechanisms, and applications for biomanufacturing of platform chemicals and fuels

In living cells, alpha-ketoacid decarboxylases (KDCs, EC 4.1.1.-) are a class of enzymes that convert alpha-ketoacids into aldehydes through decarboxylation. These aldehydes serve as either drop-in chemicals or precursors for the biosynthesis of alcohols, carboxylic acids, esters, and alkanes. These compounds play crucial roles in cellular metabolism and fitness and the bioeconomy, facilitating the sustainable and renewable biomanufacturing of platform chemicals and fuels. This review explores the diversity and classification of KDCs, detailing their structures, mechanisms, and functions. We highlight recent advancements in repurposing KDCs to enhance their efficiency and robustness for biomanufacturing. Additionally, we present modular KDC-dependent metabolic pathways for the microbial biosynthesis of aldehydes, alcohols, carboxylic acids, esters, and alkanes. Lastly, we discuss recent developments in the modular cell engineering technology that can potentially be applied to harness the diversity of KDC-dependent pathways for biomanufacturing platform chemicals and fuels.

09 BIOMASS FUELS↗
DOE OSTI2020

Towards renewable flavors, fragrances, and beyond

Esters demand a large space of unique molecules with broad range of applications as flavors, fragrances, pharmaceuticals, cosmetics, green solvents, and advanced biofuels. Global demand of natural esters in food, household cleaner, personal care, and perfume industries is increasing while the ester supply from natural sources has been limited. Development of novel microbial cell factories for ester production from renewable feedstocks can potentially provide an alternative and sustainable source of natural esters and hence help fulfill growing demand. In this work, we highlight recent advances in microbial production of esters and provide perspectives for improving its economic feasibility. As the field matures, microbial ester production platforms will enable renewable and sustainable production of flavors, fragrances, and open new market opportunities beyond what nature can offer.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1984

Engineering model system study for a regenerative fuel cell: Study report

Key design issues of the regenerative fuel cell system concept were studied and a design definition of an alkaline electrolyte based engineering model system or low Earth orbit missions was completed. Definition of key design issues for a regenerative fuel cell system include gaseous reactant storage, shared heat exchangers and high pressure pumps. A power flow diagram for the 75 kW initial space station and the impact of different regenerative fuel cell modular sizes on the total 5 year to orbit weight and volume are determined. System characteristics, an isometric drawing, component sizes and mass and energy balances are determined for the 10 kW engineering model system. An open loop regenerative fuel cell concept is considered for integration of the energy storage system with the life support system of the space station. Technical problems and their solutions, pacing technologies and required developments and demonstrations for the regenerative fuel cell system are defined.

Chang, B. J.↗
DOE OSTI2021

Designer membraneless organelles sequester native factors for control of cell behavior

Subcellular compartmentalization of macromolecules increases flux and prevents inhibitory interactions to control biochemical reactions. Inspired by this functionality, we sought to build designer compartments that function as hubs to regulate the flow of information through cellular control systems. Here we report a synthetic membraneless organelle platform to control endogenous cellular activities through sequestration and insulation of native proteins. We engineer and express a disordered protein scaffold to assemble micron size condensates and recruit endogenous clients via genomic tagging with high-affinity dimerization motifs. By relocalizing up to ninety percent of a targeted enzymes to synthetic condensates, we efficiently control cellular behaviors, including proliferation, division, and cytoskeletal organization. Further, we demonstrate multiple strategies for controlled cargo release from condensates to switch cells between functional states. These synthetic organelles offer a powerful and generalizable approach to modularly control cell decision-making in a variety of model systems with broad applications for cellular engineering.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2023

Sneaking in SpyCatcher using cell penetrating peptides for in vivo imaging

Abstract In vivo imaging of protein complexes is a powerful method for understanding the underlying biological function of these key biomolecules. Though the engineering of small, high affinity nanobodies have become more prevalent, the off-rates of these tags may result in incomplete or partial labeling of proteins in live cells. The SpyCatcher003 and SpyTag split protein system allow for irreversible, covalent binding to a short target peptide unlike nanobody-affinity based probes. However, delivering these tags into a cell without disrupting its normal function is a key challenge. Cell penetrating peptides (CPPs) are short peptide sequences that facilitate the transduction of otherwise membrane-impermeable ‘cargo’ , such as proteins, into cells. Here we report on our efforts to engineer and characterize CPP-SpyCatcher003 fusions as modular imaging probes. We selected three CPPs, CUPID, Pentratin, and pVEC, to engineer fusion protein probes for superresolution microscopy, with the aim to eliminate prior permeabilization treatments that could introduce imaging artifacts. We find that fusing the CPP sequences to SpyCatcher003 resulted in dimer and multimer formation as determined by size exclusion chromatography, dynamic light scattering, and SDS resistant dimers on SDS-PAGE gels. By isolating and labeling the monomeric forms of the engineered protein, we show these constructs retained their ability to bind SpyTag and all three CPP sequences remain membrane active, as assessed by CD spectroscopy in the presence of SDS detergent. Using fluorescence and super resolution Lattice structured illumination microscopy (Lattice SIM) imaging we show that the CPPs did not enhance uptake of SpyCatcher by E. coli, however with Caulobacter crescentus cells, we show that Penetratin, and to a lesser degree CUPID, does enhance uptake. Our results demonstrate the ability of the CPP-SpyCatcher003 to label targets within living cells, providing the groundwork for using split protein systems for targeted in vivo imaging.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Controlling selectivity of modular microbial biosynthesis of butyryl-CoA-derived designer esters

Short-chain esters have broad utility as flavors, fragrances, solvents, and biofuels. Controlling selectivity of ester microbial biosynthesis has been an outstanding metabolic engineering problem. In this study, we enabled the de novo fermentative microbial biosynthesis of butyryl-CoA-derived designer esters (e.g., butyl acetate, ethyl butyrate, butyl butyrate) in Escherichia coli with controllable selectivity. Using the modular design principles, we generated the butyryl-CoA-derived ester pathways as exchangeable production modules compatible with an engineered chassis cell for anaerobic production of designer esters. We designed these modules derived from an acyl-CoA submodule (e.g., acetyl-CoA, butyryl-CoA), an alcohol submodule (e.g., ethanol, butanol), a cofactor regeneration submodule (e.g., NADH), and an alcohol acetyltransferase (AAT) submodule (e.g., ATF1, SAAT) for rapid module construction and optimization by manipulating replication (e.g., plasmid copy number), transcription (e.g., promoters), translation (e.g., codon optimization), pathway enzymes, and pathway induction conditions. To further enhance production of designer esters with high selectivity, we systematically screened various strategies of protein solubilization using protein fusion tags and chaperones to improve the soluble expression of multiple pathway enzymes. Finally, our engineered ester-producing strains could achieve 19-fold increase in butyl acetate production (0.64 g/L, 96% selectivity), 6-fold increase in ethyl butyrate production (0.41 g/L, 86% selectivity), and 13-fold increase in butyl butyrate production (0.45 g/L, 54% selectivity) as compared to the initial strains. Altogether, this study presented a generalizable framework to engineer modular microbial platforms for anaerobic production of butyryl-CoA-derived designer esters from renewable feedstocks.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Development of a Genome-Scale Metabolic Model of Clostridium thermocellum and Its Applications for Integration of Multi-Omics Datasets and Computational Strain Design

Solving environmental and social challenges such as climate change requires a shift from our current non-renewable manufacturing model to a sustainable bioeconomy. To lower carbon emissions in the production of fuels and chemicals, plant biomass feedstocks can replace petroleum using microorganisms as biocatalysts. The anaerobic thermophile Clostridium thermocellum is a promising bacterium for bioconversion due to its capability to efficiently degrade lignocellulosic biomass. However, the complex metabolism of C. thermocellum is not fully understood, hindering metabolic engineering to achieve high titers, rates, and yields of targeted molecules. In this study, we developed an updated genome-scale metabolic model of C. thermocellum that accounts for recent metabolic findings, has improved prediction accuracy, and is standard-conformant to ensure easy reproducibility. We illustrated two applications of the developed model. We first formulated a multi-omics integration protocol and used it to understand redox metabolism and potential bottlenecks in biofuel (e.g., ethanol) production in C. thermocellum. Second, we used the metabolic model to design modular cells for efficient production of alcohols and esters with broad applications as flavors, fragrances, solvents, and fuels. The proposed designs not only feature intuitive push-and-pull metabolic engineering strategies, but also present novel manipulations around important central metabolic branch-points. We anticipate the developed genome-scale metabolic model will provide a useful tool for system analysis of C. thermocellum metabolism to fundamentally understand its physiology and guide metabolic engineering strategies to rapidly generate modular production strains for effective biosynthesis of biofuels and biochemicals from lignocellulosic biomass.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1981

Study of multi-megawatt technology needs for photovoltaic space power systems. Volume 1: Executive summary

Possible missions requiring multimegawatt photovoltaic space power systems in the 1990's time frame and associated power system technology needs are examined. The following concepts for photovoltaic power approaches are considered: planar arrays, concentrating arrays, hybrid systems using Rankine engines, thermophotovoltaic and AC/DC power management approaches, battery, fuel cell, flywheel energy storage, and interactions with the electrical ion engine injection and stationkeeping system. The levels of modularity for efficient, safe, constructable, serviceable, and cost effective system design are analyzed, and the benefits of alternate approaches developed. Both manned low Earth orbit and unmanned geosynchronous Earth orbit applications were examined for technological development. Technology developments applicable to power systems which appear to have benefits independent of the absolute power level are suggested.

Peterson, D. M.↗
NASA NTRS2014

A Modular Framework for Modeling Hardware Elements in Distributed Engine Control Systems

Progress toward the implementation of distributed engine control in an aerospace application may be accelerated through the development of a hardware-in-the-loop (HIL) system for testing new control architectures and hardware outside of a physical test cell environment. One component required in an HIL simulation system is a high-fidelity model of the control platform: sensors, actuators, and the control law. The control system developed for the Commercial Modular Aero-Propulsion System Simulation 40k (C-MAPSS40k) provides a verifiable baseline for development of a model for simulating a distributed control architecture. This distributed controller model will contain enhanced hardware models, capturing the dynamics of the transducer and the effects of data processing, and a model of the controller network. A multilevel framework is presented that establishes three sets of interfaces in the control platform: communication with the engine (through sensors and actuators), communication between hardware and controller (over a network), and the physical connections within individual pieces of hardware. This introduces modularity at each level of the model, encouraging collaboration in the development and testing of various control schemes or hardware designs. At the hardware level, this modularity is leveraged through the creation of a Simulink(R) library containing blocks for constructing smart transducer models complying with the IEEE 1451 specification. These hardware models were incorporated in a distributed version of the baseline C-MAPSS40k controller and simulations were run to compare the performance of the two models. The overall tracking ability differed only due to quantization effects in the feedback measurements in the distributed controller. Additionally, it was also found that the added complexity of the smart transducer models did not prevent real-time operation of the distributed controller model, a requirement of an HIL system.

numerical simulation↗
NASA NTRS2015

A Modular Framework for Modeling Hardware Elements in Distributed Engine Control Systems

Progress toward the implementation of distributed engine control in an aerospace application may be accelerated through the development of a hardware-in-the-loop (HIL) system for testing new control architectures and hardware outside of a physical test cell environment. One component required in an HIL simulation system is a high-fidelity model of the control platform: sensors, actuators, and the control law. The control system developed for the Commercial Modular Aero-Propulsion System Simulation 40k (C-MAPSS40k) provides a verifiable baseline for development of a model for simulating a distributed control architecture. This distributed controller model will contain enhanced hardware models, capturing the dynamics of the transducer and the effects of data processing, and a model of the controller network. A multilevel framework is presented that establishes three sets of interfaces in the control platform: communication with the engine (through sensors and actuators), communication between hardware and controller (over a network), and the physical connections within individual pieces of hardware. This introduces modularity at each level of the model, encouraging collaboration in the development and testing of various control schemes or hardware designs. At the hardware level, this modularity is leveraged through the creation of a SimulinkR library containing blocks for constructing smart transducer models complying with the IEEE 1451 specification. These hardware models were incorporated in a distributed version of the baseline C-MAPSS40k controller and simulations were run to compare the performance of the two models. The overall tracking ability differed only due to quantization effects in the feedback measurements in the distributed controller. Additionally, it was also found that the added complexity of the smart transducer models did not prevent real-time operation of the distributed controller model, a requirement of an HIL system.

propulsion simulation↗
NASA NTRS2014

A Modular Framework for Modeling Hardware Elements in Distributed Engine Control Systems

Progress toward the implementation of distributed engine control in an aerospace application may be accelerated through the development of a hardware-in-the-loop (HIL) system for testing new control architectures and hardware outside of a physical test cell environment. One component required in an HIL simulation system is a high-fidelity model of the control platform: sensors, actuators, and the control law. The control system developed for the Commercial Modular Aero-Propulsion System Simulation 40k (40,000 pound force thrust) (C-MAPSS40k) provides a verifiable baseline for development of a model for simulating a distributed control architecture. This distributed controller model will contain enhanced hardware models, capturing the dynamics of the transducer and the effects of data processing, and a model of the controller network. A multilevel framework is presented that establishes three sets of interfaces in the control platform: communication with the engine (through sensors and actuators), communication between hardware and controller (over a network), and the physical connections within individual pieces of hardware. This introduces modularity at each level of the model, encouraging collaboration in the development and testing of various control schemes or hardware designs. At the hardware level, this modularity is leveraged through the creation of a Simulink (R) library containing blocks for constructing smart transducer models complying with the IEEE 1451 specification. These hardware models were incorporated in a distributed version of the baseline C-MAPSS40k controller and simulations were run to compare the performance of the two models. The overall tracking ability differed only due to quantization effects in the feedback measurements in the distributed controller. Additionally, it was also found that the added complexity of the smart transducer models did not prevent real-time operation of the distributed controller model, a requirement of an HIL system.

numerical simulation↗
DOE OSTI2020

In vitro prototyping of limonene biosynthesis using cell-free protein synthesis

Metabolic engineering of microorganisms to produce sustainable chemicals has emerged as an important part of the global bioeconomy. Unfortunately, efforts to design and engineer microbial cell factories are challenging because design-build-test cycles, iterations of re-engineering organisms to test and optimize new sets of enzymes, are slow. To alleviate this challenge, we demonstrate a cell-free approach termed in vitro Prototyping and Rapid Optimization of Biosynthetic Enzymes (or iPROBE). In iPROBE, a large number of pathway combinations can be rapidly built and optimized. The key idea is to use cell-free protein synthesis (CFPS) to manufacture pathway enzymes in separate reactions that are then mixed to modularly assemble multiple, distinct biosynthetic pathways. As a model, we apply our approach to the 9-step heterologous enzyme pathway to limonene in extracts from Escherichia coli. In iterative cycles of design, we studied the impact of 54 enzyme homologs, multiple enzyme levels, and cofactor concentrations on pathway performance. In total, we screened over 150 unique sets of enzymes in 580 unique pathway conditions to increase limonene production in 24 h from 0.2 to 4.5 mM (23–610 mg/L). Finally, to demonstrate the modularity of this pathway, we also synthesized the biofuel precursors pinene and bisabolene. Finally, we anticipate that iPROBE will accelerate design-build-test cycles for metabolic engineering, enabling data-driven multiplexed cell-free methods for testing large combinations of biosynthetic enzymes to inform cellular design.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Modular cell-free expression plasmids to accelerate biological design in cells

Industrial biotechnology aims to produce high-value products from renewable resources. This can be challenging because model microorganisms—organisms that are easy to use like Escherichia coli—often lack the machinery required to utilize desired feedstocks like lignocellulosic biomass or syngas. Non-model organisms, such as Clostridium, are industrially proven and have desirable metabolic features but have several hurdles to mainstream use. Namely, these species grow more slowly than conventional laboratory microbes, and genetic tools for engineering them are far less prevalent. To address these hurdles for accelerating cellular design, cell-free synthetic biology has matured as an approach for characterizing non-model organisms and rapidly testing metabolic pathways in vitro. Unfortunately, cell-free systems can require specialized DNA architectures with minimal regulation that are not compatible with cellular expression. In this work, we develop a modular vector system that allows for T7 expression of desired enzymes for cell-free expression and direct Golden Gate assembly into Clostridium expression vectors. Utilizing the Joint Genome Institute’s DNA Synthesis Community Science Program, we designed and synthesized these plasmids and genes required for our projects allowing us to shuttle DNA easily between our in vitro and in vivo experiments. We next validated that these vectors were sufficient for cell-free expression of functional enzymes, performing on par with the previous state-of-the-art. Lastly, we demonstrated automated six-part DNA assemblies for Clostridium autoethanogenum expression with efficiencies ranging from 68% to 90%. We anticipate this system of plasmids will enable a framework for facile testing of biosynthetic pathways in vitro and in vivo by shortening development cycles.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2002

SMEX-Lite Modular Solar Array Architecture

For the most part, Goddard solar arrays have been custom designs that are unique to each mission. The solar panel design has been frozen prior to issuing an RFP for their procurement. There has typically been 6-9 months between RFP release and contract award, followed by an additional 24 months for performance of the contract. For Small Explorer (SMEX) missions, with three years between mission definition and launch, this has been a significant problem. The SMEX solar panels have been sufficiently small that the contract performance period has been reduced to 12-15 months. The bulk of this time is used up in the final design definition and fabrication of flight solar cell assemblies. Even so, it has been virtually impossible to have the spacecraft design at a level of maturity sufficient to freeze the solar panel geometry and release the RFP in time to avoid schedule problems with integrating the solar panels to the spacecraft. With that in mind, the SMEX-Lite project team developed a modular architecture for the assembly of solar arrays to greatly reduce the cost and schedule associated with the development of a mission- specific solar array. In the modular architecture, solar cells are fabricated onto small substrate panels. This modular panel (approximately 8.5" x 17" in this case) becomes the building block for constructing solar arrays for multiple missions with varying power requirements and geometrical arrangements. The mechanical framework that holds these modules together as a solar array is the only mission-unique design, changing in size and shape as required for each mission. There are several advantages to this approach. First, the typical solar array development cycle requires a mission unique design, procurement, and qualification including a custom qualification panel. With the modular architecture, a single qualification of the SMEX-Lite modules and the associated mechanical framework in a typical configuration provided a qualification by similarity to multiple missions. It then becomes possible to procure solar array modules in advance of mission definition and respond quickly and inexpensively to a selected mission's unique requirements. The solar array modular architecture allows the procurement of solar array modules before the array geometry has been frozen. This reduces the effect of procurement lead-time on the mission integration and test flow by as much as 50%. Second, by spreading the non-recurring costs over multiple missions, the cost per unit area is also reduced. In the case of the SMEX-Lite procurement, this reduction was by about one third of the cost per unit area compared to previous SMEX mission-unique procurements. Third, the modular architecture greatly facilitates the infusion of new solar cell technologies into flight programs as these technologies become available. New solar cell technologies need only be fabricated onto a standard-sized module to be incorporated into the next available mission. The modular solar array can be flown in a mixed configuration with some new and some standard cell technologies. Since each module has its own wiring terminals, the array can be arranged as desired electrically with little impact to cost and schedule. The solar array modular architecture does impose some additional constraints on systems and subsystem engineers. First, they must work with discrete solar array modules rather than size the array to fit exactly within an available envelope. The array area is constrained to an integer multiple of the module area. Second, the modular design is optimized for space radiation and thermal environments not greatly different from a typical SMEX LEO environment. For example, a mission with a highly elliptical orbit (e.g., Polar, SMEX/FAST) would require thicker coverglasses to protect the solar cells from the more intense radiation environment.

Lyons, John↗
NASA NTRS2004

Design and Fabrication of a Two-Dimensional Superconducting Pop-up Bolometer Array

We have been developing an architecture for producing large format, two dimensional arrays of close-packed bolometers, which will enable submillimeter cameras and spectrometers to obtain images and spectra orders of magnitude faster than present instruments. The low backgrounds achieved in these instruments require very sensitive detectors with NEPs of order 5 x 10(exp -18) W/square root of Hz. Superconducting transition edge sensor bolometers can be close-packed using the Pop-up Detector (PUD) format, and SQUID multiplexers operating at the detector base temperature can be intimately coupled to them. The array unit cell is 8 x 32 pixels, using 32- element detector and multiplexer components. We have fabricated an engineering model array with this technology which features a very compact, modular approach for large format arrays. We report on the production of the 32-element components for the arrays. Planned instruments using this array architecture include the Submillimeter and Far-InfraRed Experiment (SAFIRE) on the SOFIA airborne observatory, the South Pole Imaging Fabry-Perot Interferometer (SPIFI) for the AST/RO observatory, the Millimeter Bolometer Camera for the Atacama Cosmology Telescope (MBC/ACT), and the Redshift (Z) Early Universe Spectrometer (ZEUS j.

Benford, Dominic J.↗
DOE OSTI2023

An extensible vector toolkit and parts library for advanced engineering of plant genomes

Abstract Plant biotechnology is rife with new advances in transformation and genome engineering techniques. A common requirement for delivery and coordinated expression in plant cells, however, places the design and assembly of transformation constructs at a crucial juncture as desired reagent suites grow more complex. Modular cloning principles have simplified some aspects of vector design, yet many important components remain unavailable or poorly adapted for rapid implementation in biotechnology research. Here, we describe a universal Golden Gate cloning toolkit for vector construction. The toolkit chassis is compatible with the widely accepted Phytobrick standard for genetic parts, and supports assembly of arbitrarily complex T‐DNAs through improved capacity, positional flexibility, and extensibility in comparison to extant kits. We also provision a substantial library of newly adapted Phytobricks, including regulatory elements for monocot and dicot gene expression, and coding sequences for genes of interest such as reporters, developmental regulators, and site‐specific recombinases. Finally, we use a series of dual‐luciferase assays to measure contributions to expression from promoters, terminators, and from cross‐cassette interactions attributable to enhancer elements in certain promoters. Taken together, these publicly available cloning resources can greatly accelerate the testing and deployment of new tools for plant engineering.

42 ENGINEERING↗
NASA NTRS1977

Multimission modular spacecraft parallel battery tests

An engineering evaluation of the Multimission Modular Spacecraft power supply is presented. The spacecraft has one charger with eight commandable voltage versus temperature levels. Graphs are presented which represent: (1) cell voltage limits vs. temperature; (2) charge ratio vs. temperature; (3) effect of 0.0 percent impedance unbalance at 10 C; (4) shorted cell at 10 C; and (5) discharge voltage profile at 10 C.

Palandati, C.↗