Pseudomonas putida membrane vesicles
Cellular and membrane vesicle proteomes from Pseudomonas putida KT2440 and engineered derivative strains. Proteomic quantification by diDO-IPTL (Waldbauer et al. 2017 Analytical Chemistry).
hypervesiculation↗
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Results for “strain engineering”
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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Flight investigation of the effect of transient wing response on wing strains of a twin-engine transport airplane in rough air
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PULL-UPS↗
Flight investigation of the effect of transient wing response on wing strains of a four-engine bomber airplane in rough air
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Murrow, Harold N↗
Plant photomorphogenesis and canopy growth
An important motivation for studying photomorphogenesis is to understand the relationships among plant photophysiology in canopies, canopy productivity, and agronomic yield. This understanding is essential to optimize lighting systems used for plant farming in controlled environments (CE) and for the design of genetically engineered crop strains with altered photoresponses. This article provides an overview of some basic principles of plant photomorphogenesis in canopies and discusses their implications for (1) scaling up information on plant photophysiology from individual plants in CE to whole canopies in the field, and (2) designing lighting conditions to increase plant productivity in CE used for agronomic purposes (e.g. space farming in CE Life Support Systems). We concentrate on the visible (lambda between 400 and 700 nm) and far-infrared (FR; lambda greater than 700 nm) spectral regions, since the ultraviolet (UV; 280 to 400 nm) is covered by other authors in this volume.
Ballare, Carlos L.↗
Bionutrients-1, On-Demand Production of Nutrients in Space
Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small “production pack” system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.
Hindupur, Aditya↗
BioNutrients-1: On-Demand Production of Nutrients in Space
Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small "production pack" system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.
Hindupur, Aditya↗
BioNutrients-1: Development of an On-Demand Nutrient Production System for Long-Duration Missions
Future long-duration missions beyond low-Earth orbit will require advances in food technologies to address the documented problem of degradation of vitamins and nutrients in supplied foods stored long-term. To begin to address the issue of nutrient degradation, we are developing and flight-testing a platform biomanufacturing technology for in situ production of target nutrients. This technology is being tested over a five-year duration on the International Space Station (ISS). As part of the BioNutrients-1 project we have developed an on-demand system for the production of two carotenoids, β-carotene and zeaxanthin, by genetically engineering distinct strains of Saccharomyces cerevisiae, more commonly known as baker’s yeast. The on-orbit nutrient production packs contain a desiccated yeast strain and edible growth substrate. Once hydrated, the contents of the production packs are intended to grow and produce a desired amount of ready-to-consume nutrients. In this current version the production packs will not be consumed and future missions will require an inactivation of microorganisms before consumption. In addition to the on-orbit hydration of the production packs a series of valuable microorganisms are currently being stored in stasis packs on the ISS including probiotics organisms, bacterial strains used in yogurt production, and organisms with potential use for future biomanufacturing. Analysis of returned ISS stasis packs and ground controls will include multi-omics studies and provide insight into long-term survival of organisms stored in a space environment. Both stasis packs and hydrated production packs will be intermittently returned to Earth for analysis. Preliminary data from long-term storage studies of stasis packs stored on the ISS for 47 days versus their ground control counterparts have shown no significant difference in viability. Currently no production packs have been processed.
Yeast↗
BioNutrients: Microbial on-demand production of short shelf-life micronutrients in space
Long duration deep space missions pose a significant challenge when considering adequate nutrition for human spaceflight. Many micronutrients supplied in pre-packaged foods for low Earth orbit flights have a short shelf life and will degrade over multi-year missions. One strategy to mitigate the risk of micronutrient malnutrition is to grow fresh foods to supplement a pre-packaged diet. The BioNutrients project aims to develop systems for growing edible microorganisms that are genetically engineered to produce short shelf life micronutrients. BioNutrients-1 (BN-1) is a five-year spaceflight project currently underway on the International Space Station (ISS) that tests long-term storage and production of two carotenoids, β-carotene and zeaxanthin, in genetically engineered yeast strains. BN-1 on-orbit and ground control activities indicate that the nutrient production system will be stable for at least two years in ambient storage conditions. BN-1 is also testing the effects of long-term ISS storage on viability and gene expression using “stasis packs” containing a series of microorganisms useful in food and biomanufacturing processes. Stasis pack analysis is distinguishing microorganisms with full viability after two years of storage from other species that lose viability rapidly. In parallel, BioNutrients-2 (BN-2) is being developed as a six-month flight experiment that will expand the range of foods produced on ISS to include yogurt, kefir, and probiotic yeast. BN-2 will improve upon hardware design by transitioning from a hard-shell polycarbonate bioreactor to a flexible fluorinated ethylene propylene (FEP) bag. This change reduces the mass and volume required to store production packs. The long-term goal of BioNutrients is to develop the capability for rapid and safe delivery of multiple micronutrients in single-use production packs. We expect synthetic biology approaches similar to BioNutrients will play an important role for astronaut health in future NASA missions to the Moon and Mars.
Synthetic Biology↗
Environmental and High-Strain Rate effects on composites for engine applications
The Lewis Research Center is conducting a series of programs intended to investigate and develop the application of composite materials to structural components for turbojet engines. A significant part of that effort is directed to establishing resistance, defect growth, and strain rate characteristics of composite materials over the wide range of environmental and load conditions found in commercial turbojet engine operations. Both analytical and experimental efforts are involved.
Chamis, C. C.↗
Adaptive laboratory evolution and genetic engineering improved terephthalate utilization in Pseudomonas putida KT2440
Poly(ethylene terephthalate) (PET) is one of the most ubiquitous plastics and can be depolymerized through biological and chemo-catalytic routes to its constituent monomers, terephthalic acid (TPA) and ethylene glycol (EG). TPA and EG can be re-synthesized into PET for closed-loop recycling or microbially converted into higher-value products for open-loop recycling. Here, in this study, we expand on our previous efforts engineering and applying Pseudomonas putida KT2440 for PET conversion by employing adaptive laboratory evolution (ALE) to improve TPA catabolism. Three P. putida strains with varying degrees of metabolic engineering for EG catabolism underwent an automation-enabled ALE campaign on TPA, a TPA and EG mixture, and glucose as a control. ALE increased the growth rate on TPA and TPA-EG mixtures by 4.1- and 3.5-fold, respectively, in approximately 350 generations. Evolved isolates were collected at the midpoints and endpoints of 39 independent ALE experiments, and growth rates were increased by 0.15 and 0.20 h -1 on TPA and a TPA-EG, respectively, in the best performing isolates. Whole-genome re-sequencing identified multiple converged mutations, including loss-of-function mutations to global regulators gacS, gacA, and turA along with large duplication and intergenic deletion events that impacted the heterologously-expressed tphAB II catabolic genes. Reverse engineering of these targets confirmed causality, and a strain with all three regulators deleted and second copies of tphAB II and tpaK displayed improved TPA utilization compared to the base strain. Taken together, an iterative strain engineering process involving heterologous pathway engineering, ALE, whole genome sequencing, and genome editing identified five genetic interventions that improve P. putida growth on TPA, aimed at developing enhanced whole-cell biocatalysts for PET upcycling.
36 MATERIALS SCIENCE↗
Effect of Angle of Incidence of Second-Stage Vane Assembly on Third-Stage Compressor-Blade Vibration and Engine Performance
Strain-gages were used to measure blade vibrations causing failures in the third stage of a production 11-stage axial-flow compressor. After the serious third-stage vibration was detected, a series of investigations were conducted with second-stage vane assemblies of varying angles of incidence. Curves presented herein show the effect of varying the angle of incidence of second-stage vane assembly on third-stage rotor-blade vibration amplitude and engine performance. A minimum vibration amplitude was obtained without greatly affecting the engine performance with a second-stage vane assembly of 9deg. greater angle of incidence than the assembly normally furnished with the engine.
Meyer, Andre J., Jr.↗
Leveraging a synthetic biology approach to enhance BCG-mediated expansion of Vγ9Vδ2 T cells
There is an urgent need to develop a more efficacious anti-tuberculosis vaccine as the current live-attenuated vaccine strain BCG fails to prevent pulmonary infection in adults. In this study, we leverage a synthetic biology approach to engineer BCG to produce more (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), an intermediate of bacterial—but not host—isoprenoid biosynthesis via the methylerythritol phosphate (MEP) pathway. HMBPP strongly activates and expands Vγ9Vδ2 T cells, which are unique to higher-order primates and protect against Mycobacterium tuberculosis infection. BCG has been engineered to produce specific ligands and antigens to some success; in contrast, our strategy exploits a self-nonself recognition mechanism in the host via HMBPP sensing, which has not been attempted before. To inform the design of our recombinant strains, we performed synteny analyses of >63 mycobacterial species and found that isoprenoid biosynthetic genes are not operonic across all the 356 surveyed genomes, but some genes are frequently found in pairs. Thus, we generated synthetic loci with the goal of specifically overproducing HMBPP and tested the ability of these engineered strains to induce human Vγ9Vδ2 expansion in an in vitro stimulation assay. We found that BCG expressing a synthetic MEP locus significantly enhanced Vγ9Vδ2 T cell expansion over the wild-type vaccine strain, and overexpression of the HMBPP synthase GcpE alone potently induced Vγ9Vδ2 T cell expansion with no downregulation of other pathway genes. Together these engineered strains present two successful strategies to accumulate HMBPP and overcome feedback inhibition of the MEP pathway.
59 BASIC BIOLOGICAL SCIENCES↗
Environmental and high strain rate effects on composites for engine applications
The complex environmental and loading conditions experienced by many turbine engine components impose severe durability and damage tolerance requirements for component materials. Programs are being conducted to establish the structural performance of composite materials under anticipated engine operating environments. A description is presented of the results obtained in connection with several of these programs. A comparison of predicted and measured hygrothermal effects is considered along with hygrothermal effects on defect growth, high strain rate effects on composite mechanical properties, dynamic stress intensity factors for composites, and the indentation laws for composite impact analysis.
Chamis, C. C.↗
Calculation of Dynamic Loads Due to Random Vibration Environments in Rocket Engine Systems
An important part of rocket engine design is the calculation of random dynamic loads resulting from internal engine "self-induced" sources. These loads are random in nature and can greatly influence the weight of many engine components. Several methodologies for calculating random loads are discussed and then compared to test results using a dynamic testbed consisting of a 60K thrust engine. The engine was tested in a free-free condition with known random force inputs from shakers attached to three locations near the main noise sources on the engine. Accelerations and strains were measured at several critical locations on the engines and then compared to the analytical results using two different random response methodologies.
Christensen, Eric R.↗
Strain gage system evaluation program
A program was conducted to determine the reliability of various strain gage systems when applied to rotating compressor blades in an aircraft gas turbine engine. A survey of current technology strain gage systems was conducted to provide a basis for selecting candidate systems for evaluation. Testing and evaluation was conducted in an F 100 engine. Sixty strain gage systems of seven different designs were installed on the first and third stages of an F 100 engine fan. Nineteen strain gage failures occurred during 62 hours of engine operation, for a survival rate of 68 percent. Of the failures, 16 occurred at blade-to-disk leadwire jumps (84 percent), two at a leadwire splice (11 percent), and one at a gage splice (5 percent). Effects of erosion, temperature, G-loading, and stress levels are discussed. Results of a post-test analysis of the individual components of each strain gage system are presented.
Dolleris, G. W.↗
Environmental and high strain rate effects on composites for engine applications
Previously cited in issue 13, p. 2034, Accession no. A82-30118
Chamis, C. C.↗
Leveraging CRISPR Cas9 RNPs and Cre- loxP in Picochlorum celeri for generation of field deployable strains and selection marker recycling
As new highly productive strains of algae are discovered and developed to meet the energy, chemical, and food requirements of the future, genetic engineering of those strains in a manner that yields deployable transformants is paramount. This study introduces the novel CRoxP ($\underline{\textrm{C}}$$\textrm{as9}$ $\underline{\textrm{R}}$$\textrm{NPs}$ coupled with an inducible $\underline{\textrm{CR}}$$\textrm{e}$-$\textrm{l}\underline{\textrm{oxP}}$) system for rapid generation of marker- and transgene-free strains of Picochlorum celeri. The CRoxP system allows reuse of selection markers without Cas9 expression in vivo, eliminating many of the bottlenecks associated with conventional CRISPR Cas9 use for precise genome editing. In P. celeri, transformants were generated with a turnaround time as short as 21 days between transformation and being ready for another round of transformation with the same selection marker by using the CRoxP system. As a use-case for CRoxP, depigmented strains of P. celeri were generated by multiplexed Cas9 disruption of major LHCII genes followed by either a second round of LHCII targeting, or knockout of an LHCI gene. One transformant tested in flask culture (R6) exhibited similar biomass production to the wild type with 46% less Chl a + b on a biomass basis. In photobioreactors and under diel light simulating a solar day, a transformant (LhcBM31) exhibited 34 g AFDW m –2 d –1 with 54% less Chl a + b on a biomass basis vs. wild type.
09 BIOMASS FUELS↗
Benchmark notch test for life prediction
Aircraft gas turbine engine components are subjected to severe stress, temperature, and environmental conditions. Economic and reliabilty demands have prompted inordinate effort in development of analytic methods to predict stresses and strains in aircraft engines. There remains, however, the need to check or verify these analytical methodologies against actual experimental data measurements. The laser interferometric strain displacement gage was recognized as having the potential to accomplish this task and was employed in this program. The actual strains incurred at the root of a discontinuity in cyclically loaded test samples subjected to inelastic deformation at high temperature where creep deformation readily occur were measured. The steady-state, cyclic stress-strain response at the root of the discontinuity in the tested samples was analyzed for comparison with the measured results. A comprehensive set of local notch root strain measurements for a variety of load patterns in an Inconel 718 notch specimen at 649 C (1200 F) was obtained and documented using the laser interferometric strain displacement gage.
Domas, P. A.↗