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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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All sourcesNASA NTRSDOE OSTI
NASA NTRS1996

A bioreactor system for the nitrogen loop in a Controlled Ecological Life Support System

As space missions become longer in duration, the need to recycle waste into useful compounds rises dramatically. This problem can be addressed by the development of Controlled Ecological Life Support Systems (CELSS) (i.e., Engineered Closed/Controlled Eco-Systems (ECCES)), consisting of human and plant modules. One of the waste streams leaving the human module is urine. In addition to the reclamation of water from urine, recovery of the nitrogen is important because it is an essential nutrient for the plant module. A 3-step biological process for the recycling of nitrogenous waste (urea) is proposed. A packed-bed bioreactor system for this purpose was modeled, and the issues of reaction step segregation, reactor type and volume, support particle size, and pressure drop were addressed. Based on minimization of volume, a bioreactor system consisting of a plug flow immobilized urease reactor, a completely mixed flow immobilized cell reactor to convert ammonia to nitrite, and a plug flow immobilized cell reactor to produce nitrate from nitrite is recommended. It is apparent that this 3-step bioprocess meets the requirements for space applications.

NASA Discipline Number 61-10↗
NASA NTRS2008

Polymer Coats Leads on Implantable Medical Device

Langley Research Center s Soluble Imide (LaRC-SI) was discovered by accident. While researching resins and adhesives for advanced composites for high-speed aircraft, Robert Bryant, a Langley engineer, noticed that one of the polymers he was working with did not behave as predicted. After putting the compound through a two-stage controlled chemical reaction, expecting it to precipitate as a powder after the second stage, he was surprised to see that the compound remained soluble. This novel characteristic ended up making this polymer a very significant finding, eventually leading Bryant and his team to win several NASA technology awards, and an "R&D 100" award. The unique feature of this compound is the way that it lends itself to easy processing. Most polyimides (members of a group of remarkably strong and incredibly heat- and chemical-resistant polymers) require complex curing cycles before they are usable. LaRC-SI remains soluble in its final form, so no further chemical processing is required to produce final materials, like thin films and varnishes. Since producing LaRC-SI does not require complex manufacturing techniques, it has been processed into useful forms for a variety of applications, including mechanical parts, magnetic components, ceramics, adhesives, composites, flexible circuits, multilayer printed circuits, and coatings on fiber optics, wires, and metals. Bryant s team was, at the time, heavily involved with the aircraft polymer project and could not afford to further develop the polymer resin. Believing it was worth further exploration, though, he developed a plan for funding development and submitted it to Langley s chief scientist, who endorsed the experimentation. Bryant then left the high-speed civil transport project to develop LaRC-SI. The result is an extremely tough, lightweight thermoplastic that is not only solvent-resistant, but also has the ability to withstand temperature ranges from cryogenic levels to above 200 C. The thermoplastic s unique characteristics lend it to many commercial applications; uses that Bryant believed would ultimately benefit industry and the Nation. "LaRC-SI," he explains, "is a product created in a government laboratory, funded with money from the tax-paying public. What we discovered helps further the economic competitiveness of the United States, and it was our goal to initiate the technology transfer process to ensure that our work benefited the widest range of people." Several NASA centers, including Langley, have explored methods for using LaRC-SI in a number of applications from radiation shielding and as an adhesive to uses involving replacement of conventional rigid circuit boards. In the commercial realm, LaRC-SI can now be found in several commercial products, including the thin-layer composite unimorph ferroelectric driver and sensor (THUNDER) piezoelectric actuator, another "R&D 100" award winner (Spinoff 2005).

Source record↗
NASA NTRS2015

3D Photocatalytic Air Processor for Dramatic Reduction of Life Support Mass and Complexity

To dramatically reduce the cost and risk of CO2 management systems in future extended missions, we have conducted preliminary studies on the materials and device development for advanced "artificial photosynthesis" reaction systems termed the High Tortuosity PhotoElectroChemical (HTPEC) system. Our Phase I studies have demonstrated that HTPEC operates in much the same way a tree would function, namely directly contacting the cabin air with a photocatalyst in the presence of light and water (as humidity) to immediately conduct the process of CO2 reduction to O2 and useful, "tunable" carbon products. This eliminates many of the inefficiencies associated with current ISS CO2 management systems. We have laid the solid foundation for Phase II work to employ novel and efficient reactor geometries, lighting approaches, 3D manufacturing methods and in-house grown novel catalytic materials.The primary objective of the proposed work is to demonstrate the scientific and engineering foundation for light-activated, compact devices capable of converting CO2 to O2 and mission-relevant carbon compounds. The proposed HTPEC CO2 management system will demonstrate a novel pathway with high efficiency and reliability in a compact, lightweight reactor architecture. The proposed HTPEC air processing concept can be developed in multiple architectures, such as centralized processing as well as "artificial leaves" distributed throughout the cabin that utilize pre-existing cabin ventilation (wind). Additionally, HTPEC can be deployed with spectrally tunable collection channels for selectable product generation. HTPEC employs light as its only energy source to remove and convert waste CO2 using a non-toxic composite catalyst.We have demonstrated in the Phase I studies the production, tunability and robustness of the novel composite catalysts following the preliminary work in the Chen laboratory. Additionally, we have designed, fabricated and tested all components of HTPEC device with active materials, including flow modeling to optimize flow mixing and pressure drop as well as the production of ethylene and other larger hydrocarbons. To best determine how this technology could be implemented, we also performed system integration optimization and trade studies. This includes parameters such as mass, volume, power in relation to selected mission configurations, CO2 delivery methods and light source/delivery approaches.

Artificial Photosynthesis↗
NASA NTRS2023

Evolutionary Flexibility and Rigidity in the Bacterial Methylerythritol Phosphate (Mep) Pathway

Terpenoids are a diverse class of compounds with wide-ranging uses including as industrial solvents, pharmaceuticals, and fragrances. Efforts to produce terpenoids sustainably by engineering microbes for fermentation are ongoing, but industrial production still largely relies on nonrenewable sources. The methylerythritol phosphate (MEP) pathway generates terpenoid precursor molecules and includes the enzyme Dxs and two iron–sulfur cluster enzymes: IspG and IspH. IspG and IspH are rate limiting-enzymes of the MEP pathway but are challenging for metabolic engineering because they require iron–sulfur cluster biogenesis and an ongoing supply of reducing equivalents to function. Therefore, identifying novel alternatives to IspG and IspH has been an on-going effort to aid in metabolic engineering of terpenoid biosynthesis. We report here an analysis of the evolutionary diversity of terpenoid biosynthesis strategies as a resource for exploration of alternative terpenoid biosynthesis pathways. Using comparative genomics, we surveyed a database of 4,400 diverse bacterial species and found that some may have evolved alternatives to the first enzyme in the pathway, Dxs making it evolutionarily flexible. In contrast, we found that IspG and IspH are evolutionarily rigid because we could not identify any species that appear to have enzymatic routes that circumvent these enzymes. The ever-growing repository of sequenced bacterial genomes has great potential to provide metabolic engineers with alternative metabolic pathway solutions. With the current state of knowledge, we found that enzymes IspG and IspH are evolutionarily indispensable which informs both metabolic engineering efforts and our understanding of the evolution of terpenoid biosynthesis pathways.

Bailey Marshall↗
DOE OSTI2022

6.26 Low Cost Basalt Fiber for Automotive Applications

Vehicle lightweighting is an essential component to the automotive industry to improve fuel economy of internal combustion engine (ICE) vehicles to meet ever improving emission standards and to improve the range of electric vehicles (EV). Composite materials offer high specific modulus and specific strength, which makes them appealing for these light weighting efforts. Sheet molding compounds (SMC) are particularly interesting from an automotive perspective because of the relatively low cost and high volume of producing SMC parts. Traditionally, composite materials for automotive application are glass fiber reinforced because of the attractive price - performance ratio, but basalt fibers are a cost and recycling competitive reinforcement alternative in this market. The aim of this project was to examine the feasibility of utilizing basalt fiber for automotive applications. More specifically, an effort was made to examine different fiber sizings on basalt fiber combined with vinyl-ester (VE) resin, and their performance as part of an SMC process. In addition to offering vehicle lightweighting with fiber reinforced polymer composites, basalt fiber is a fully recyclable material and thus supports the IACMI technical goal of: Demonstrate that the technology is capable at a sufficient scale for >80% recyclability or reuse of fiber reinforced polymer composites in five years into useful components with projected cost and quality at commercial scale competitive with virgin materials on a pathway to 95% recyclability or reuse starting in ten years. Three different fiber sizings were applied to a continuous roving of basalt fiber and compared to a traditional Electrical/Chemical Resistance (E-CR) glass fiber that is typically used in these types of applications. Fiber tows were examined for Loss on Ignition percentage (LOI%), Tex, and tow strength. Some sizings clearly outperformed others, and the ability to process these fibers on a pilot scale SMC line was demonstrated. A test plan for the manufacturing and mechanical testing of SMC panels was developed. This work continues outside the time frame allocated for this project. When this work is completed, it will be added to this report and posted as Appendix C. Glass fiber reinforced SMC materials have already proven feasible as a light weighting method for traditionally steel parts like the Volkswagen (VW) Atlas lift-gate (Figure 1); this project team is seeking the feasibility of basalt fiber as a drop-in replacement for glass fiber reinforced SMC. Sizing development for basalt fibers has proven that the mechanical properties are better than E-glass and closer to S-glass, which makes it an interesting material for SMC applications. Better mechanical properties translate to less material needed to achieve load case requirement for target applications. The business case has already been demonstrated for 100,000 parts per year of glass fiber reinforced SMC Atlas lift-gates compared to traditional steel manufacturing processes. Reduced overhead and assembly costs are offset by glass fiber SMC higher cost per kg beyond 100,000 parts per year, which is still a relatively low volume for the automotive industry. For basalt fiber reinforced SMC to become feasible for automotive applications, the price-performance ratio has to be precisely determined. Based on the mechanical performance it is possible to establish a range of applications and technical solutions in which the potential of basalt SMC can be utilized, while the price of the material can be used to compile the business case for such applications. Based on these business cases and the sustainability indicators, glass fiber reinforcement (or other) materials can be directly substituted. Volkswagen’s commitment to reducing carbon emissions cannot be understated. Basalt fiber shows promise of reducing the carbon footprint in SMC materials, especially if sizing optimizations can be made with thermoplastic based SMC. To fully realize the value of basalt fiber reinforced materials, a lifecycle cost analysis should be performed on basalt’s production and recycling, and then compared against E-glass. From this assessment, a true judgement can be made on the commercialization potential of this material. Figure 1. Example of Fiber Reinforced Polymer Composite Liftgate As a conclusion, we can state that Mafic basalt fiber is not a direct replacement for E-glass or E-CR glass based on price, but should be considered a technical solution when E-glass does not provide adequate performance in a composite design and S-glass, aramid and carbon fibers are too costly. Mafic basalt fiber can be placed on the high-performance fiber spectrum next to S-glass for performance but at one third the price. It should be considered for more technically challenging structural designs wherein the performance can demonstrate 20-25% performance enhancement over E-glass to elicit more strength or a weight reduction. Both Michelman and Mafic produce thermoplastic sizings which, in combination with Nylon and polypropylene resin and fibers, can further advance high speed composite implementations while maintaining an eco-friendly manufacturing process.

36 MATERIALS SCIENCE↗
DOE OSTI2020

Engineering Surface Oxygenated Functionalities on Commercial Carbon toward Ultrafast Sodium Storage in Ether-Based Electrolytes

The pursuit of a high-capacity anode material has been urgently required for commercializing sodium-ion batteries with a high energy density and an improved working safety. In the absence of thermodynamically stable sodium intercalated compounds with graphite, constructing nanostructures with expanded interlayer distances is still the mainstream option for developing high-performance carbonaceous anodes. In this regard, a surface-functionalized and pore-forming strategy through a facile CO 2 thermal etching route was rationally adopted to engineer negligible oxygenated functionalities on commercial carbon for boosting the sodium storage process. Benefitted from the abundant ionic/electronic pathways and more active reaction sites in the microporous structure with noticeable pseudocapacitive behaviors, the functionalized porous carbon could achieve a highly reversible capacity of 505 mA h g -1 at 50 mA g -1 , an excellent rate performance of 181 mA h g -1 at 16,000 mA g -1 , and an exceptional rate cycle stability of 176 mA h g -1 at 3200 mA g -1 over 1000 cycles. These outstanding electrochemical properties should be ascribed to a synergistic mechanism, fully utilizing the graphitic and amorphous structures for synchronous intercalations of sodium ions and solvated sodium ion compounds, respectively. Finally, the controllable generation and evolution of a robust but thin solid electrolyte interphase film with the emergence of obvious capacitive reactions on the defective surface, favoring the rapid migration of sodium ions and solvated species, also contribute to a remarkable electrochemical performance of this porous carbon black.

25 ENERGY STORAGE↗
DOE OSTI2022

Volatile organic compound emissions from 4D printing: Effects of material composition and external stimulus

4D printing of stimuli-responsive materials extends 3D printing by enabling the fabricated structures to transform their shapes and properties over time in response to external stimuli. Numerous research efforts have been dedicated to developing new smart materials, enhancing material printability, and ensuring time-evolving properties. Meanwhile, the use of smart materials and external stimuli in 4D printing has introduced the possibility of air emissions that can potentially deteriorate the indoor air quality at the workplace and pose continuous health hazards to users during the production and use phases. These potential air emissions caused by 4D printing have not yet been assessed in current literature, leading to unknown occupational hazards and human health effects. This study focuses on stereolithography-based 4D printing with constrained thermo-mechanics and builds an emission model to quantify the volatile organic compound emissions from printing, shape programming, and shape recovery stages. The established model mathematically links the emission characteristics with material compositional design and stimuli-response mechanisms. Additionally, shape fixity and recovery abilities are considered to analyze the trade-off between the air emissions and stimuli-response performance of 4D printed parts. Case study results suggest that the methacrylate-based thermo-responsive material with higher glass transition temperature leads to higher air emissions, surpassing the permissible exposure level in the indoor environment. By altering the thermo-temporal conditions, a 61.29% reduction in emission yield can be achieved while ensuring a satisfactory shape memory performance.

36 MATERIALS SCIENCE↗
DOE OSTI2026

Interfacial Hydrogen-Bond Dynamics in Transition Metal Compounds

Understanding how water behaves when confined within atomic layers of active transition-metal carbides, nitrides, and carbonitrides is essential for uncovering the fundamental principles needed to engineer solid–liquid interfaces at the atomic scale. Yet, how lattice element chemistry and surface termination groups collectively regulate the structure and mobility of such interlayer water remains poorly understood. Here, we present a composition-controlled investigation of interlayer water dynamics in layered transition-metal nitride, carbide, and carbonitride systems using a systematic integration of quasi-elastic neutron scattering (QENS), ab initio molecular dynamics (AIMD) simulations, and density functional theory (DFT) calculations. QENS measurements show that nitride-rich systems host mobile, translationally diffusing water with thermally activated self-diffusion coefficients on the order of 10 –10 m 2 s –1 , whereas mixed C/N lattices confine water to localized, nontranslational motion that is insensitive to temperature. AIMD and DFT reveal that lattice C/N chemistry and surface functional group composition reshape the first hydration layer by modulating the surface electronic structure and termination-dependent hydrogen-bond networks, leading to pronounced differences in water ordering and thermal resilience. On the other hand, fully carbide systems exhibit intermediate behavior, highlighting that water mobility is not primarily controlled by the hydration level alone but by the coupling between lattice composition and surface chemistry. Overall, this study establishes how surface chemistry and lattice composition jointly control interfacial hydrogen bond dynamics, offering a mechanistic framework for designing transition-metal layered materials with tailored interfacial transport properties.

Hydration↗
DOE OSTI2022

Considering Strain Variation and Non-Type Strains for Yeast Metabolic Engineering Applications

A variety of yeast species have been considered ideal hosts for metabolic engineering to produce value-added chemicals, including the model organism Saccharomyces cerevisiae, as well as non-conventional yeasts including Yarrowia lipolytica, Kluyveromyces marxianus, and Pichia pastoris. However, the metabolic capacity of these microbes is not simply dictated or implied by genus or species alone. Within the same species, yeast strains can display distinct variations in their phenotypes and metabolism, which affect the performance of introduced pathways and the production of interesting compounds. Moreover, it is unclear how this metabolic potential corresponds to function upon rewiring these organisms. These reports thus point out a new consideration for successful metabolic engineering, specifically: what are the best strains to utilize and how does one achieve effective metabolic engineering? Understanding such questions will accelerate the host selection and optimization process for generating yeast cell factories. In this review, we survey recent advances in studying yeast strain variations and utilizing non-type strains in pathway production and metabolic engineering applications. Additionally, we highlight the importance of employing portable methods for metabolic rewiring to best access this metabolic diversity. Finally, we conclude by highlighting the importance of considering strain diversity in metabolic engineering applications.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Towards engineering ectomycorrhization into switchgrass bioenergy crops via a lectin receptor‐like kinase

Summary Soil‐borne microbes can establish compatible relationships with host plants, providing a large variety of nutritive and protective compounds in exchange for photosynthesized sugars. However, the molecular mechanisms mediating the establishment of these beneficial relationships remain unclear. Our previous genetic mapping and whole‐genome resequencing studies identified a gene deletion event of a Populus trichocarpa lectin receptor‐like kinase gene PtLecRLK1 in Populus deltoides that was associated with poor‐root colonization by the ectomycorrhizal fungus Laccaria bicolor . By introducing PtLecRLK1 into a perennial grass known to be a non‐host of L. bicolor , switchgrass ( Panicum virgatum L.), we found that L. bicolor colonizes ZmUbipro‐PtLecRLK1 transgenic switchgrass roots, which illustrates that the introduction of PtLecRLK1 has the potential to convert a non‐host to a host of L. bicolor . Furthermore, transcriptomic and proteomic analyses on inoculated‐transgenic switchgrass roots revealed genes/proteins overrepresented in the compatible interaction and underrepresented in the pathogenic defence pathway, consistent with the view that pathogenic defence response is down‐regulated during compatible interaction. Metabolomic profiling revealed that root colonization in the transgenic switchgrass was associated with an increase in N‐containing metabolites and a decrease in organic acids, sugars, and aromatic hydroxycinnamate conjugates, which are often seen in the early steps of establishing compatible interactions. These studies illustrate that PtLecRLK1 is able to render a plant susceptible to colonization by the ectomycorrhizal fungus L. bicolor and shed light on engineering mycorrhizal symbiosis into a non‐host to enhance plant productivity and fitness on marginal lands.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1994

Advanced diesel engine component development program, tasks 4-14

This report summarizes the Advanced Diesel Engine Component Development (ADECD) Program to develop and demonstrate critical technology needed to advance the heavy-duty low heat rejection engine concept. Major development activities reported are the design, analysis, and fabrication of monolithic ceramic components; vapor phase and solid film lubrication; electrohydraulic valve actuation; and high pressure common rail injection. An advanced single cylinder test bed was fabricated as a laboratory tool in studying these advanced technologies. This test bed simulates the reciprocator for a system having no cooling system, turbo compounding, Rankine bottoming cycle, common rail injection, and variable valve actuation to achieve fuel consumption of 160 g/kW-hr (.26 lb/hp-hr). The advanced concepts were successfully integrated into the test engine. All ceramic components met their functional and reliability requirements. The firedeck, cast-in-place ports, valves, valve guides, piston cap, and piston ring were made from silicon nitride. Breakthroughs required to implement a 'ceramic' engine included the fabrication of air-gap cylinder heads, elimination of compression gaskets, machining of ceramic valve seats within the ceramic firedeck, fabrication of cast-in-place ceramic port liners, implementation of vapor phase lubrication, and elimination of the engine coolant system. Silicon nitride valves were successfully developed to meet several production abuse test requirements and incorporated into the test bed with a ceramic valve guide and solid film lubrication. The ADECD cylinder head features ceramic port shields to increase insulation and exhaust energy recovery. The combustion chamber includes a ceramic firedeck and piston cap. The tribological challenge posed by top ring reversal temperatures of 550 C was met through the development of vapor phase lubrication using tricresyl phosphate at the ring-liner interface. A solenoid-controlled, variable valve actuation system that eliminated the conventional camshaft was demonstrated on the test bed. High pressure fuel injection via a common rail system was also developed to reduce particulate emissions.

Kaushal, Tony S.↗
DOE OSTI2024

Harnessing Fermentation May Enhance the Performance of Biological Sulfate-Reducing Bioreactors

Biological sulfate reduction (BSR) represents a promising strategy for bioremediation of sulfate-rich waste streams, yet the impact of metabolic interactions on performance is largely unexplored. Here, genome-resolved metagenomics was used to characterize 17 microbial communities in reactors treating synthetic sulfate-contaminated solutions. Reactors were supplemented with lactate or acetate and a small amount of fermentable substrate. Of the 163 genomes representing all the abundant bacteria, 130 encode 321 NiFe and FeFe hydrogenases and all genomes of the 22 sulfate-reducing microorganisms (SRM) encode genes for H 2 uptake. We observed lactate oxidation solely in the first packed bed reactor zone, with propionate and acetate oxidation in the middle and predominantly acetate oxidation in the effluent zone. The energetics of these reactions are very different, yet sulfate reduction kinetics were unaffected by the type of electron donor available. We hypothesize that the comparable rates, despite the typically slow growth of SRM on acetate, are a result of the consumption of H 2 generated by fermentation. This is supported by the sustained performance of a predominantly acetate-supplemented stirred tank reactor dominated by diverse fermentative bacteria encoding FeFe hydrogenase genes and SRM capable of acetate and hydrogen consumption and CO 2 assimilation. Thus, addition of fermentable substrates to stimulate syntrophic relationships may improve the performance of BSR reactors supplemented with inexpensive acetate.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Atomically dispersed Ag on δ-MnO 2 via cation vacancy trapping for toluene catalytic oxidation

The regulation of metal–support interaction (MSI) and the improvement of noble metal utilization are the frontiers of volatile organic compound (VOC) catalytic oxidation. Here, we report a H 2 O 2 -assisted synthesis process for trapping Ag + onto the engineered Mn vacancies of δ-MnO 2 for the catalytic oxidation of toluene. Due to the strong MSI between Ag and δ-MnO 2 , the monoatomic Ag + anchored on cation vacancies forms longer and weakened Ag–O bonds compared with Ag 2 O, which can extra promote the formation of oxygen vacancies and the activation of adjacent lattice oxygen in Ag–O–Mn units compared with regular nanoparticle counterparts. It is beneficial to the steps of methyl dehydrogenation, demethylation, and benzene ring cleavage. The catalyst yields excellent toluene oxidation activity, enhanced turnover frequency in terms of Ag, and good stability. Finally, we propose in this study a facile design strategy for fabricating stably atomically dispersed metal sites, as well as clarify the structure–activity relationships during toluene catalytic oxidation, especially the important effect of single-atom Ag on the adjacent local structure.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
NASA NTRS1927

The Heinkel commercial airplane, H.D. 40

The H.D. 40 has a compound structure in which metal has been used at stress points. It is powered by one BMW IV 460 H.P. engine.

AIRPLANES - HEINKEL H D 40 (Ger )↗
NASA NTRS2011

Performance Characterization of Loctite (Registered Trademark) 242 and 271 Liquid Locking Compounds (LLCs) as a Secondary Locking Feature for International Space Station (ISS) Fasteners

Several International Space Station (ISS) hardware components use Loctite (and other polymer based liquid locking compounds (LLCs)) as a means of meeting the secondary (redundant) locking feature requirement for fasteners. The primary locking method is the fastener preload, with the application of the Loctite compound which when cured is intended to resist preload reduction. The reliability of these compounds has been questioned due to a number of failures during ground testing. The ISS Program Manager requested the NASA Engineering and Safety Center (NESC) to characterize and quantify sensitivities of Loctite being used as a secondary locking feature. The findings and recommendations provided in this investigation apply to the anaerobic LLCs Loctite 242 and 271. No other anaerobic LLCs were evaluated for this investigation. This document contains the findings and recommendations of the NESC investigation

Dube, Michael J.↗
NASA NTRS2005

Electroporation System for Sterilizing Water

A prototype of an electroporation system for sterilizing wastewater or drinking water has been developed. In electroporation, applied electric fields cause transient and/or permanent changes in the porosities of living cells. Electroporation at lower field strengths can be exploited to increase the efficiency of chemical disinfection (as in chlorination). Electroporation at higher field strengths is capable of inactivating and even killing bacteria and other pathogens, without use of chemicals. Hence, electroporation is at least a partial alternative to chlorination. The transient changes that occur in micro-organisms at lower electric-field strengths include significantly increased uptake of ions and molecules. Such increased uptake makes it possible to achieve disinfection at lower doses of chemicals (e.g., chlorine or ozone) than would otherwise be needed. Lower doses translate to lower costs and reduced concentrations of such carcinogenic chemical byproducts as trichloromethane. Higher electric fields cause cell membranes to lose semipermeability and thereby become unable to function as selective osmotic barriers between the cells and the environment. This loss of function is the cause of the cell death at higher electric-field intensities. Experimental evidence does not indicate cell lysis but, rather, combined leaking of cell proteins out of the cells as well as invasion of foreign chemical compounds into the cells. The concept of electroporation is not new: it has been applied in molecular biology and genetic engineering for decades. However, the laboratory-scale electroporators used heretofore have been built around small (400-microliter) cuvettes, partly because the smallness facilitates the generation of electric fields of sufficient magnitude to cause electroporation. Moreover, most laboratory- scale electroporators have been designed for testing static water. In contrast, the treatment cell in the present system is much larger and features a flow-through geometry, such that electric fields strong enough to effect 99.9- percent disinfection can be applied to water flowing in a pipe.

Schlager, Kenneth J.↗
DOE OSTI2022

Improving growth of Cupriavidus necator H16 on formate using adaptive laboratory evolution-informed engineering

Conversion of CO 2 to value-added products presents an opportunity to reduce GHG emissions while generating revenue. Formate, which can be generated by the electrochemical reduction of CO 2 , has been proposed as a promising intermediate compound for microbial upgrading. Here we present progress towards improving the soil bacterium Cupriavidus necator H16, which is capable of growing on formate as its sole source of carbon and energy using the Calvin-Benson-Bassham (CBB) cycle, as a host for formate utilization. Using adaptive laboratory evolution, we generated several isolates that exhibited faster growth rates on formate. The genomes of these isolates were sequenced, and resulting mutations were systematically reintroduced by metabolic engineering, to identify those that improved growth. Further, the metabolic impact of several mutations was investigated further using RNA-seq transcriptomics. We found that deletion of a transcriptional regulator implicated in quorum sensing, PhcA, reduced expression of several operons and led to improved growth on formate. Growth was also improved by deleting large genomic regions present on the extrachromosomal megaplasmid pHG1, particularly two hydrogenase operons and the megaplasmid CBB operon, one of two copies present in the genome. Based on these findings, we generated a rationally engineered ..delta..phcA and megaplasmid-deficient strain that exhibited a 24% faster maximum growth rate on formate. Moreover, this strain achieved a 7% growth rate improvement on succinate and a 19% increase on fructose, demonstrating the broad utility of microbial genome reduction. This strain has the potential to serve as an improved microbial chassis for biological conversion of formate to value-added products.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Development of Gelatin-Coated Hydrogel Microspheres for Novel Bioink Design: A Crosslinker Study

The development of vascularized tissue is a substantial challenge within the field of tissue engineering and regenerative medicine. Studies have shown that positively-charged microspheres exhibit dual-functions: (1) facilitation of vascularization and (2) controlled release of bioactive compounds. In this study, gelatin-coated microspheres were produced and processed with either EDC or transglutaminase, two crosslinkers. The results indicated that the processing stages did not significantly impact the size of the microspheres. EDC and transglutaminase had different effects on surface morphology and microsphere stability in a simulated colonic environment. Incorporation of EGM and TGM into bioink did not negatively impact bioprintability (as indicated by density and kinematic viscosity), and the microspheres had a uniform distribution within the scaffold. These microspheres show great potential for tissue engineering applications.

60 APPLIED LIFE SCIENCES↗