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At least 1,135 records · Page 63
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Characterizing and Engineering Trehalose Biosynthesis Pathways to Improve Acetate Tolerance in Pseudomonas putida KT2440

Alkaline pretreatment can depolymerize lignin into a diverse mixture of monomers and smaller compounds, enabling bioconversion of these compounds to value-added chemicals in engineered bacterial hosts such as Pseudomonas putida KT2440. Acetate and salts comprise approximately 8% and 32% of the lignin stream, respectively. High concentrations of these components can cause cell death in P. putida, so lignin-derived streams must be fed in lower amounts to decrease stress. Investigation of the mechanisms of bacterial tolerance to chemicals in lignin-rich substrates is therefore required to overcome this obstacle in lignin bioconversion. Previous work has shown that trehalose, an endogenously produced disaccharide, is involved in tolerance to heat, cold, and osmotic stress, as well as implicated in acid stress in other bacteria. As such, trehalose biosynthesis presents a useful target to increase acetate and osmotic tolerance in P. putida. This work investigates the native TreSA, TreSB, and TreY/TreZ trehalose biosynthesis pathways in this bacterial strain as well as the OtsAB pathway of Sphingobium sp. SYK-6. Overexpression of the OtsAB pathway in P. putida increased tolerance to acetate relative to wild-type. Additionally, functional knockouts of treSA, treSB, treY, and treZ in P. putida showed that interruption of any trehalose biosynthesis pathway, and, in the case of TreY/TreZ, interruption of the pathway at any point, significantly hindered growth in acetate as well as on glucose and LB broth. Each trehalose biosynthesis pathway was then engineered for overexpression in P. putida to determine whether overexpression of these genes conferred additional tolerance to acetate. The TreSA, TreSB, OtsA, and OtsB enzymes were also expressed and purified to determine the reaction rates and kinetics of these pathways. The hypothetical glycoside hydrolase family 15 protein, encoded in the gene located in between otsB and otsA in the Sphingobium operon, was purified as well to investigate its potential involvement in the OtsAB pathway. The findings of this work illuminated the mechanisms of trehalose biosynthesis in P. putida and identified genetic targets to improve strain tolerance to common components of alkaline-pretreated lignin streams. These findings could ultimately improve yields of desired products from lignin in engineered strains of P. putida.

alkaline pretreatment↗
NASA NTRS1992

A probabilistic approach to the evaluation of fatigue damage in a space propulsion system injector element

The fatigue damage of a space propulsion system component is computed using probabilistic structural analysis methods. The analysis takes into account the variations in static and dynamic loads, the uncertainty in structural damping, and the scatter in material fatigue resistance. The key elements of the probabilistic approach include: (1) a numerical engine model for describing the global component interface loads consistent with engine balance, (2) models for computing the local mode boundary conditions on the component, (3) a static structural analysis model that captures the strains at a damage critical location as a function of engine performance variables, (4) a finite element model for assessment of the random amplitude stress response due to random base and pressure excitations with uncertain power and correlation length, and (5) advanced first order reliability methods for computing the probabilities associated with the fatigue damage.

Rajagopal, K. R.↗
DOE OSTI2016

Onset of Plasticity via Relaxation Analysis (OPRA)

In crystalline metals and alloys, plasticity occurs due to the movement of mobile dislocations and the yield stress for engineering applications is traditionally quantified based on strain. The onset of irreversible plasticity or “yielding” is generally identified by a deviation from linearity in the stress-strain plot or by some standard convention such as 0.2 % offset strain relative to the “linear elastic response”. In the present work, we introduce a new methodology for the determination of the true yield point based on stress relaxation. We show experimentally that this determination is self-consistent in nature and, as such, provides an objective observation of the very onset of plastic flow. Lastly, our designation for yielding is no longer related to the shape of the stress-strain curve but instead reflects the earliest signature of the activation of concerted irreversible dislocation motion in a test specimen under increasing load.

36 MATERIALS SCIENCE↗
DOE OSTI2019

Performance of a ferritic/martensitic steel for nuclear reactor applications fabricated using additive manufacturing

Ferritic/martensitic (FM) steels are being targeted for use in a range of advanced reactor concepts as cladding and structural components. FM steels for nuclear reactor applications have historically been produced using traditional methods (e.g., casting and forging), but recently, additive manufacturing processes have become of interest for making FM-based components. Here, the laser-blown-powder additive manufacturing process was used to fabricate an FM steel, HT9, followed by microstructural and mechanical performance evaluations to determine the viability of future use of additive manufacturing for FM-based component fabrication. Results showed that the as-built condition formed a layered structure with alternating layers of δ-ferrite and martensite, which resulted in anisotropic engineering and true-stress, true-strain mechanical performance. Post-build normalizing and tempering treatments alerted the prior austenite grain size and precipitate distributions, and drove the mechanical performance to near-isotropic properties that mimic wrought-processed properties. Overall, the resulting microstructures in all conditions were rationalized in the context of multi-pass welding and the synergies are discussed.

36 MATERIALS SCIENCE↗
DOE OSTI2018

Engineering a spermidine biosynthetic pathway in Clostridium thermocellum results in increased resistance to furans and increased ethanol production

Polyamines are low molecular weight aliphatic nitrogen compounds found ubiquitously in microorganisms, plants, and animals. Spermidine is a common polyamine that plays a role in stabilizing chromatin, DNA replication, transcription, translation, as well as the regulation of cell growth and apoptosis in eukaryotes. Amines are also associated with defense to a number of environmental stresses including elevated temperature and have been shown to be involved in tolerance to fermentation inhibitors such as furan derivatives and acetic acid in Saccharomyces cerevisiae. While the tolerance and detoxifying mechanisms have been intensively studied, metabolic engineering efforts to construct tolerant and resistant strains have been few. Here, in this work, we show that exogenously added spermidine confers enhanced tolerance to furans and acetic acid in the Gram-positive bacterium, Clostridium thermocellum . Deletion of the endogenous spermidine synthase resulted in a severe growth defect and hypersensitivity to both furans and acetic acid. Exogenously added spermidine rescued all three phenotypes. Overexpression of the endogenous spermidine synthase resulted in increased tolerance to these compounds without added spermidine. Increased tolerance to these fermentation inhibitors will facilitate the use of C. thermocellum , one of the most cellulolytic of all known bacterial species, for the production of fuels from plant biomass substrates.

Biofuels↗
DOE OSTI2021

Synthesis, antibiotic structure–activity relationships, and cellulose dissolution studies of new room-temperature ionic liquids derived from lignin

Ionic liquids (ILs) are promising pretreatment solvents for lignocellulosic biomass, but are largely prepared from petroleum precursors. Benzaldehydes from depolymerized lignin, such as vanillin, syringaldehyde, and 4-methoxy benzaldehyde, represent renewable feedstocks for the synthesis of ionic liquids. We herein report syntheses of novel lignin-derived ionic liquids, with extended N -alkyl chains, and examine their melting points, cellulose dissolution capacities, and toxicity profiles against Daphnia magna and E. coli strain 1A1. The latter organism has been engineered to produce isoprenol, a drop-in biofuel and precursor for commodity chemicals. The new N,N -diethyl and N,N -dipropyl methyl benzylammonium ILs were liquids at room temperature, showing 75–100 °C decreased melting points as compared to their N,N,N -trimethyl benzylammonium analog. Extension of N -alkyl chains also increased antibacterial activity threefold, while ionic liquids prepared from vanillin showed 2- to 4-fold lower toxicity as compared to those prepared from syringaldehyde and 4-methoxybenzaldehyde. The trend of antibacterial activity for anions of lignin-derived ILs was found to be methanesulfonate < acetate < hydroxide. Microcrystalline cellulose dissolution, from 2 to 4 wt% after 20 min at 100 °C, was observed in all new ILs using light microscopy and IR spectroscopy. Ionic liquids prepared from H-, S- and G-lignin oxidation products provided differential cytotoxic activity against E. coli and D. magna , suggesting these compounds could be tailored for application specificity within a biorefinery.

09 BIOMASS FUELS↗
DOE OSTI2014

Increase in ethanol yield via elimination of lactate production in an ethanol-tolerant mutant of Clostridium thermocellum

Large-scale production of lignocellulosic biofuel is a potential solution to sustainably meet global energy needs. One-step consolidated bioprocessing (CBP) is a potentially advantageous approach for the production of biofuels, but requires an organism capable of hydrolyzing biomass to sugars and fermenting the sugars to ethanol at commercially viable titers and yields. Clostridium thermocellum, a thermophilic anaerobe, can ferment cellulosic biomass to ethanol and organic acids, but low yield, low titer, and ethanol sensitivity remain barriers to industrial production. Here, we deleted the hypoxanthine phosphoribosyltransferase gene in ethanol tolerant strain of C. thermocellum adhE*(EA) in order to allow use of previously developed gene deletion tools, then deleted lactate dehydrogenase (ldh) to redirect carbon flux towards ethanol. Upon deletion of ldh, the adhE*(EA) ldh strain produced 30% more ethanol than wild type on minimal medium. The adhE*(EA) ldh strain retained tolerance to 5% v/v ethanol, resulting in an ethanol tolerant platform strain of C. thermocellum for future metabolic engineering efforts.

Biswas, Ranjita↗
DOE OSTI2012

Dying for Good: Virus-Bacterium Biofilm Co-evolution Enhances Environmental Fitness

Commonly used in biotechnology applications, filamentous M13 phage are non-lytic viruses that infect E. coli and other bacteria, with the potential to promote horizontal gene transfer in natural populations with synthetic biology implications for engineering community systems. Using the E. coli strain TG1, we have investigated how a selective pressure involving elevated levels of toxic chromate, mimicking that found in some superfund sites, alters population dynamics following infection with either wild-type M13 phage or an M13-phage encoding a chromate reductase (Gh-ChrR) capable of the reductive immobilization of chromate (ie, M13-phageGh-ChrR). In the absence of a selective pressure, M13-phage infection results in a reduction in bacterial growth rate; in comparison, in the presence of chromate there are substantial increases in both cellular killing and biomass formation following infection of E. coli strain TG1with M13-phageGh-ChrR that is dependent on chromate-reductase activity. These results are discussed in terms of community structures that facilitate lateral gene transfer of beneficial traits that enhance phage replication, infectivity, and stability against environmental change.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1987

Nonlinear heat transfer and structural analyses of SSME turbine blades

Three-dimensional nonlinear finite-element heat transfer and structural analyses were performed for the first stage high-pressure fuel turbopump blade of the space shuttle main engine (SSME). Directionally solidified (DS) MAR-M 246 material properties were considered for the analyses. Analytical conditions were based on a typical test stand engine cycle. Blade temperature and stress-strain histories were calculated using MARC finite-element computer code. The study was undertaken to assess the structural response of an SSME turbine blade and to gain greater understanding of blade damage mechanisms, convective cooling effects, and the thermal-mechanical effects.

Abdul-Aziz, A.↗
NASA NTRS1990

New heterojunction LWIR detector options

Researchers investigate the heterojunction internal photoemission (HIP) approach that potentially offers long wavelength infrared (LWIR) photovoltaic detector performance (single pixel) that is competitive with the best of other approaches being considered. Most significantly, this approach offers a relatively simple device technology that promises producible and uniform FPA's. Researchers emphasize an exciting process based on intervalence band absorption. They investigate both III-V and Si-based heterojunctions grown by molecular beam epitaxy (MBE) in which the barrier can be tailored to the desired cutoff wavelength. In addition, MBE allows one to optimize the device structure with precise control of doping profiles and layer thicknesses, and perform band structure engineering by control of composition and heterojunction strain. Researchers also consider free carrier absorption in heterojunctions. Acceptable absorption coefficients can be achieved in very heavily n(exp +) doped semiconductor layers (approx. equals 10(exp 20)cm(exp -3). However, in this case the appreciable filling of conduction band states leads to a Schottky-like photoresponse with a gradual (quadratic) turn-on above threshold. A more satisfactory approach would be to use p(exp +) doping so that with the higher density of states in the heavy hole valence band there would be a narrow band of occupied states. This gives the desirable effect of a more rapid (linear) turn-on above threshold. Unfortunately, the higher hole effective mass also reduces (inversely) the free carrier absorption. For this and other reasons, the intervalence band absorption process looks much more promising.

Maserjian, Joseph↗
NASA NTRS1991

Hierarchic Extensions in the Static and Dynamic Analysis of Elastic Beams

Approximate solutions of static and dynamic beam problems by the p-version of the finite element method are investigated. Within a hierarchy of engineering beam idealizations, rigorous formulations of the strain and kinetic energies for straight and circular beam elements are presented. These formulations include rotating coordinate system effects and geometric nonlinearities to allow for the evaluation of vertical axis wind turbines, the motivating problem for this research. Hierarchic finite element spaces, based on extensions of the polynomial orders used to approximate the displacement variables, are constructed. The developed models are implemented into a general purpose computer program for evaluation. Quality control procedures are examined for a diverse set of sample problems. These procedures include estimating discretization errors in energy norm and natural frequencies, performing static and dynamic equilibrium checks, observing convergence for qualities of interest, and comparison with more exacting theories and experimental data. It is demonstrated that p-extensions produce exponential rates of convergence in the approximation of strain energy and natural frequencies for the class of problems investigated.

Watson, Robert A.↗
NASA NTRS1992

Creep fatigue life prediction for engine hot section materials (isotropic)

A series of high temperature strain controlled fatigue tests have been completed to study the effects of thermomechanical fatigue, multiaxial loading, reactive environments, and imposed mean stresses. The baseline alloy used in these tests was cast B1900+Hf (with and without coatings); a small number of tests of wrought INCO 718 are also included. A strong path dependence was demonstrated during the thermomechanical fatigue testing, using in-phase, out-phase, and non-proportional (elliptical and 'dogleg') strain-temperature cycles. The multiaxial tests also demonstrated cycle path to be a significant variable, using both proportional and non-proportional tension-torsion loading. Environmental screening tests were conducted in moderate pressure oxygen and purified argon; the oxygen reduced the specimen lives by two, while the argon testing produced ambiguous data. Both NiCoCrAlY overlay and diffusion aluminide coatings were evaluated under isothermal and TMF conditions; in general, the lives of the coated specimens were higher that those of uncoated specimens. Controlled mean stress TMF tests showed that small mean stress changes could change initiation lives by orders of magnitude; these results are not conservatively predicted using traditional linear damage summation rules. Microstructures were evaluated using optical, SEM and TEM methods.

Nelson, R. S.↗
DOE OSTI2023

Assessing continuum plasticity postulates with grain stress and local strain measurements in triaxially compressed sand

Critical state and continuum plasticity theories have been used in research and engineering practice in soil and rock mechanics for decades. These theories rely on postulated relationships between material stresses and strains. Some classical postulates include coaxiality between stress and strain rates, stress–dilatancy relationships, and kinematic assumptions in shear bands. Although numerical and experimental data have quantified the strains and grain kinematics in such experiments, little data quantifying grain stresses are available. Here, we report the first-known grain stress and local strain measurements in triaxial compression tests on synthetic quartz sands using synchrotron X-ray tomography and 3D X-ray diffraction. We use these data to examine the micromechanics of shear banding, with a focus on coaxiality, stress-dilatancy, and kinematics within bands. Our results indicate the following: 1) elevated deviatoric stress, strain, and stress ratios in shear bands throughout experiments; 2) coaxial principal compressive stresses and strains throughout samples; 3) significant contraction along shear bands; 4) vanishing volumetric strain but nonvanishing stress fluctuations throughout samples at all stages of deformation. Our results provide some of the first-known in situ stress and strain measurements able to aid in critically evaluating postulates employed in continuum plasticity and strain localization theories for sands.

42 ENGINEERING↗
NASA NTRS1984

Evaluation results of the 700 deg C Chinese strain gages

There is a continuing interest and need for resistance strain gages capable of making static strain measurements on components located in the hot section of gas turbine engines. A paper by Tsen-tai Wu describes the development and evaluation of high temperature gauges fabricated from specially developed Fe-Cr-Al-V-Ti-Y alloy wire. Several of these gages and a quantity of P12-2 ceramic adhesive were purchased for evaluation. Nine members of the aircraft turbine engine community were invited to participate in an evaluation of these gages. Each participant was sent one strain gage, a small amount of ceramic adhesive, instructions for mounting the gage on a test beam, and a set of suggestions for the experiment. Data on gage factor variation with temperature, apparent strain, and drift are discussed.

Hobart, H. F.↗
DOE OSTI2017

Metabolic engineering of Bacillus subtilis for production of D-lactic acid

Poly lactic acid (PLA) based plastics is renewable, bio–based, and biodegradable. Although present day PLA is composed of mainly L–LA, an L– and D– LA copolymer is expected to improve the quality of PLA and expand its use. To increase the number of thermotolerant microbial biocatalysts that produce D–LA, a derivative of Bacillus subtilis strain 168 that grows at 50°C was metabolically engineered. Since B. subtilis lacks a gene encoding D–lactate dehydrogenase (ldhA), five heterologous ldhA genes (B. coagulans ldhA and gldA101, and ldhA from three Lactobacillus delbrueckii) were evaluated. Corresponding D–LDHs were purified and biochemically characterized. Among these, D–LDH from L. delbrueckii subspecies bulgaricus supported the highest D…LA titer (about 1M) and productivity (2 g h –1 g cells –1 ) at 37°C (B. subtilis strain DA12). The D–LA titer at 48°C was about 0.6 M at a yield of 0.99 (g D–LA g–1glucose consumed). Strain DA12 also fermented glucose at 48°C in mineral salts medium to lactate at a yield of 0.89 g g –1 glucose and the D–lactate titer was 180 ± 4.5 mM. Furthermore, these results demonstrate the potential of B. subtilis as a platform organism for metabolic engineering for production of chemicals at 48°C that could minimize process cost.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Structurally Deformed MoS 2 for Electrochemically Stable, Thermally Resistant, and Highly Efficient Hydrogen Evolution Reaction

Abstract The emerging molybdenum disulfide (MoS 2 ) offers intriguing possibilities for realizing a transformative new catalyst for driving the hydrogen evolution reaction (HER). However, the trade‐off between catalytic activity and long‐term stability represents a formidable challenge and has not been extensively addressed. This study reports that metastable and temperature‐sensitive chemically exfoliated MoS 2 (ce‐MoS 2 ) can be made into electrochemically stable (5000 cycles), and thermally robust (300 °C) while maintaining synthetic scalability and excellent catalytic activity through physical‐transformation into 3D structurally deformed nanostructures. The dimensional transition enabled by a high throughput electrohydrodynamic process provides highly accessible, and electrochemically active surface area and facilitates efficient transport across various interfaces. Meanwhile, the hierarchically strained morphology is found to improve electronic coupling between active sites and current collecting substrates without the need for selective engineering the electronically heterogeneous interfaces. Specifically, the synergistic combination of high strain load stemmed from capillarity‐induced‐self‐crumpling and sulfur (S) vacancies intrinsic to chemical exfoliation enables simultaneous modulation of active site density and intrinsic HER activity regardless of continuous operation or elevated temperature. These results provide new insights into how catalytic activity, electrochemical‐, and thermal stability can be concurrently enhanced through the physical transformation that is reminiscent of nature, in which properties of biological materials emerge from evolved dimensional transitions.

Chen, Yen‐Chang↗
DOE OSTI2012

Analytical description of true stress-true strain curves for neutron-irradiated stainless austenitic steels

This paper summarizes the results of an investigation for the deformation hardening behaviors of neutron-irradiated stainless steels in terms of true stress( ) true strain( ) curves. It is commonly accepted that the - curves are more informative for describing plastic flow, but there are few papers devoted to using the true curves for describing constitutive behaviors of materials. This study uses the true curves obtained from stainless steel samples irradiated to doses in the range of 0 55 dpa by various means: finite element calculation, optic extensomentry, and recalculation of engineering curves. It is shown that for the strain range 0 0.6 the true curves can be well described by the Swift equation: =k ( - 0)0.5. The influence of irradiation on the parameters of the Swift equation is investigated in detail. It is found that in most cases the k-parameter of this equation is not changed significantly by irradiation. Since large data scattering was observed for the 0-parameter, a modified Swift equation =k*( - 0 2/k2)0.5 was proposed and evaluated. This equation is based on the concept of zero stress, which is, in general, close to yield stress. The relationships among k, 0, and damage dose are discussed in detail, so as to more accurately describe the true curves for irradiated stainless steels.

36 MATERIALS SCIENCE↗
DOE OSTI2023

Deletion of biofilm synthesis in Eubacterium limosum ATCC 8486 improves handling and transformation efficiency

Abstract Eubacterium limosum is an acetogenic bacterium of potential industrial relevance for its ability to efficiently metabolize a range of single carbon compounds. However, extracellular polymeric substance (EPS) produced by the type strain ATCC 8486 is a serious impediment to bioprocessing and genetic engineering. To remove these barriers, here we bioinformatically identified genes involved in EPS biosynthesis, and targeted several of the most promising candidates for inactivation, using a homologous recombination-based approach. Deletion of a single genomic region encoding homologues for epsABC, ptkA, and tmkA resulted in a strain incapable of producing EPS. This strain is significantly easier to handle by pipetting and centrifugation, and retains important wild-type phenotypes including the ability to grow on methanol and carbon dioxide and limited oxygen tolerance. Additionally, this strain is also more genetically tractable with a 2-fold increase in transformation efficiency compared to the highest previous reports. This work advances a simple, rapid protocol for gene knockouts in E. limosum using only the native homologous recombination machinery. These results will hasten the development of this organism as a workhorse for valorization of single carbon substrates, as well as facilitate exploration of its role in the human gut microbiota.

Sanford, Patrick A.↗