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At least 1,081 records · Page 60

Strain Imaging of Nanoscale Semiconductor Heterostructures with X-Ray Bragg Projection Ptychography

We report the imaging of nanoscale distributions of lattice strain and rotation in complementary components of lithographically engineered epitaxial thin film semiconductor heterostructures using synchrotron x-ray Bragg projection ptychography (BPP). We introduce a new analysis method that enables lattice rotation and out-of-plane strain to be determined independently from a single BPP phase reconstruction, and we apply it to two laterally adjacent, multiaxially stressed materials in a prototype channel device. These results quantitatively agree with mechanical modeling and demonstrate the ability of BPP to map out-of-plane lattice dilatation, a parameter critical to the performance of electronic materials.

Holt, Martin V.↗

Electroporation-Based Genetic Manipulation in Type I Methanotrophs

Methane is becoming a major candidate for a prominent carbon feedstock in the future, and the bioconversion of methane into valuable products has drawn increasing attention. In order to facilitate the use of methanotrophic organisms as industrial strains and accelerate our ability to metabolically engineer methanotrophs, simple and rapid genetic tools are needed. Electroporation is one such enabling tool, but to date it has not been successful in a group of methanotrophs of interest for the production of chemicals and fuels, the gammaproteobacterial (type I) methanotrophs. In this study, we developed electroporation techniques with a high transformation efficiency for three different type I methanotrophs: Methylomicrobium buryatense 5GB1C, Methylomonas sp. strain LW13, and Methylobacter tundripaludum 21/22. We further developed this technique in M. buryatense , a haloalkaliphilic aerobic methanotroph that demonstrates robust growth with a high carbon conversion efficiency and is well suited for industrial use for the bioconversion of methane. On the basis of the high transformation efficiency of M. buryatense , gene knockouts or integration of a foreign fragment into the chromosome can be easily achieved by direct electroporation of PCR-generated deletion or integration constructs. Moreover, site-specific recombination (FLP-FRT [FLP recombination target] recombination) and sacB counterselection systems were employed to perform marker-free manipulation, and two new antibiotics, zeocin and hygromycin, were validated to be antibiotic markers in this strain. Together, these tools facilitate the rapid genetic manipulation of M. buryatense and other type I methanotrophs, promoting the ability to perform fundamental research and industrial process development with these strains.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Conjugation-based genome engineering enables rapid prototyping and bioproduction in non-model bacteria

Abstract Non-model bacteria offer unique metabolic capabilities for sustainable bioproduction, yet their limited genetic accessibility hinders systematic strain development. Here we present conjugation-based serine recombinase-assisted genome engineering (cSAGE), a broad-host-range platform that enables predictable, iterative genomic integration in transformation-resistant bacteria. cSAGE combines conjugative DNA delivery, standardized low-copy vectors, orthogonal recombinases, and modular genetic parts to support rapid pathway assembly and cross-host benchmarking. Using purple nonsulfur bacteria as a testbed, we integrate promoter engineering, multi-payload genome modification, and genome-scale metabolic modeling to empirically evaluate host-dependent pathway performance. Applying this workflow, we identify strain-specific differences in photosynthetic conversion of lignin-derived p -coumarate to the thermoplastic precursor p -vinylphenol. By enabling genome engineering and functional comparison across diverse bacteria using a single plasmid system, cSAGE provides a general framework for non-model strain prototyping and biotransformation discovery.

Guzman, Michael S. [Department of Chemical Enginee↗

Measurement of maximum strain of steel beam structures based on average strains from vibrating wire strain gages

In structural health monitoring, the safety of steel beam structures can be assessed by comparing the measured maximum stress and the allowable stress of the beam calculated by a design code. For the case of a steel beam subjected to variable lateral loadings, many difficulties exist in measuring the maximum stress in a beam with point sensors that can measure the strain only at a local point of a beam since the location and magnitude of the maximum stress induced in a beam by the loading change. Although traditional strain sensors can measure the strain only at a local point, a vibrating wire strain gage (VWSG) that measures integrated strains over its gage length can consider the variation of strains due to variable loadings. This paper presents an estimation model to determine the maximum strains or stresses in a steel beam based on average strains measured using VWSGs. The model is derived by defining the relation between the average strains measured using VWSGs and the maximum strains of beams. The model is experimentally tested by comparing the maximum strain directly obtained from electrical strain gages and the estimated maximum strain based on the average strain from VWSGs.

42 ENGINEERING↗

A Method for Removal of Bakelite-Impregnated Wire Strain Gages

An increasing interest is being shown in the use of wire-type strain gages to measure static and dynamic stresses in aircraft-engine parts. Bakelite cement has been found satisfactory as a bonding agent for attaching the strain gages to machine parts that must operate at elevated temperatures. On many occasions, it is desired to remove Bakelite-cemented gages from the test parts for the purpose of replacing faulty gages or of returning the parts to service after strain measurements have been completed. Removal of the gages by means of scraping without prior treatment is very unsatisfactory because it is tedious and almost invariably damages the finished surface. Various solvents have been tried, but all attempts in this direction have been unsuccessful inasmuch as Bakelite cement, when properly baked, forms a polymer of very high molecular weight that resists the action of solvents. This report presents a gas-flame method of removal that is rapid and does not inure the structural part.

Kemp, Richard H.↗

Structural analysis

Hot section components of aircraft gas turbine engines are subjected to severe thermal-structural loading conditions, especially during the start-up and take-off portions of the engine cycle. The most severe and damaging stresses and strains are those induced by the steep thermal gradients induced during the start-up transient. These transient stresses and strains are also the most difficult to predict, in part because the temperature gradients and distributions are not well known or predictable, and also because the cyclic elasto-viscoplastic behavior of the materials at these extremes of temperature and strain are not well known or predictable. One element of the structures program will develop improved time-varying thermal-mechanical load models for the entire engine mission cycle from start-up to shutdown. The thermal model refinements will be consistent with those required by the structural code including considerations of mesh-point density, strain concentrations, and thermal gradients. Models will be developed for the burner liner, turbine vane and turbine blade.

Johns, R. H.↗

The development of a high temperature static strain gage system

The objective of this program is to develop electrical resistance strain gages which will permit the measurement of static strains on nickel and cobalt superalloy parts inside gas turbine engines running on a test stand. The specific goal is to develop a complete system able to make strain measurements up to plus or minus 2000 mu strain with a total error of no more than plus or minus 10 percent over a 50 hour period at 1250 K. The initial part of this work consisted of a strain gage alloy development effort in which a variety of alloys were evaluated after being prepared by drop-casting or splat cooling.

Hulse, Charles O.↗

Fiber-Optic Strain-Gage Tank Level Measurement System for Cryogenic Propellants

Measurement of tank level, particularly for cryogenic propellants, has proven to be a difficult problem. Current methods based on differential pressure, capacitance sensors, temperature sensors, etc.; do not provide sufficiently accurate or robust measurements, especially at run time. These methods are designed to measure tank-level, but when the fluids are in supercritical state, the liquid-gas interface disappears. Furthermore, there is a need for a non-intrusive measurement system; that is, the sensors should not require tank modifications and/or disturb the fluids. This paper describes a simple, but effective method to determine propellant mass by measuring very small deformations of the structure supporting the tank. Results of a laboratory study to validate the method, and experimental data from a deployed system are presented. A comparison with an existing differential pressure sensor shows that the strain gage system provides a much better quality signal across all regimes during an engine test. Experimental results also show that the use of fiber optic strain gages (FOSG) over classic foil strain gages extends the operation time (before the system becomes uncalibrated), and increases accuracy. Finally, a procedure is defined whereby measurements from the FOSG mounted on the tank supporting structure are compensated using measurements of a FOSG mounted on a reference plate and temperature measurements of the structure. Results describing the performance of a deployed system that measures tank level during propulsion tests are included.

Figueroa, Fernando↗

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↗

Tensile Behavior of Inconel X750 Sheets: Effect of Heat Treatment

X-750 is a nickel-chromium based super alloy of usefulness in a wide variety of applications. Its excellent relaxation resistance is useful for high-temperature springs and bolts; it is utilized in gas turbines, rocket engines, nuclear reactors, pressure vessels, tooling, and aircraft structures. Its performance is similar to that of Inconel alloy 600 but it is made precipitation-hardenable by additions of aluminum and titanium. It has good resistance to corrosion and oxidation along with high tensile and creep-rupture properties at temperatures up to 700 deg. C. In this study, the tensile flow behavior of the alloy was studied by performing uniaxial tensile tests under a range of temperatures and at different strain rates. To assess the microstructure-mechanical property relationships in the alloy, microstructure analysis (through the use of electron microscopy and XRD analysis) was utilized post-experiment. This paper will however focus on reporting the tensile properties and temperature and strain rate effects on the as-received alloy vs. the heat-treated alloy. (authors)

36 MATERIALS SCIENCE↗

CRAGE enables rapid activation of biosynthetic gene clusters in undomesticated bacteria

Advances in the prediction of biosynthetic gene clusters (BGCs) from genomic and metagenomic data have made the large-scale identification of candidate secondary metabolite biosynthesis pathways routine. However, most predicted BGCs cannot be successfully expressed in the limited range of expression hosts currently available, which prevents the identification and characterization of their respective products. Here we show that parallelized use of a phylogenetically and physiologically diverse panel of purpose-engineered chassis strains offers a versatile strategy for the rapid activation of BGCs. We domesticated an ensemble of 25 diverse ?-proteobacteria from 11 different genera by integrating a unified landing pad that enables efficient introduction of large BGCs into their genome. We deployed this chassis-independent recombinase-assisted genome engineering (CRAGE) strategy to express several previously posited but experimentally elusive non-ribosomal peptide synthase (NRPS) and NRPSpolyketide synthase (PKS) hybrid BGCs from Photorhabdus luminescens. Successful activation of 6 BGCs identified 21 products, including several with likely roles in host-microbe interactions. Taken together, we illustrate how the regulatory and physiological diversity naturally present across bacteria can be harnessed for the exploration of novel bioproducts and provide a sizable panel of domesticated chassis strains to enable this strategy.

applied microbiology, genetic engineering, industr↗

Rapid Design and Engineering of Smart and Secure Microbiological Systems (Final Report)

The design and application of successfully engineered biosystems requires an understanding of how engineered microbes will interact with other organisms – either as one-on-one competitors or in the context of microbial consortia. Engineering microorganisms from first principles for non-laboratory, environmental applications is inherently challenging because: (1) engineered systems tend to quickly revert back to their wild-type behaviors; and (2) these systems typically pay a price in reduced fitness, making them uncompetitive against invasive contaminating species (i.e., metabolic burden). For this project, we used a synthetic biology-based strategy to investigate the organization, control, stabilization, and destabilization of natural and engineered microbes. This approach enabled development of (1) single-strain systems capable of detecting and responding to target organisms in the environment; (2) a pipeline for refining and engineering biological constructs in new, non-model host organisms; and (3) improved systems for rapidly designing, engineering, and assaying new biological modules. This coupled approach to safeguard system design is predictable and portable across bacterial species and is focused on microbes that are part of the beneficial plant microbiome. A long-term goal beyond the proposed research is to enable the rational engineering of microbial communities based on first principles of biological design that mimic the smart performance of microorganisms observed in natural systems.

59 BASIC BIOLOGICAL SCIENCES↗

Carbon-negative production of acetone and isopropanol by gas fermentation at industrial pilot scale

Many industrial chemicals that are produced from fossil resources could be manufactured more sustainably through fermentation. In this work, we describe the development of a carbon-negative fermentation route to producing the industrially important chemicals acetone and isopropanol from abundant, low-cost waste gas feedstocks, such as industrial emissions and syngas. Using a combinatorial pathway library approach, we first mined a historical industrial strain collection for superior enzymes that we used to engineer the autotrophic acetogen Clostridium autoethanogenum. Next, we used omics analysis, kinetic modeling and cell-free prototyping to optimize flux. Finally, we scaled-up our optimized strains for continuous production at rates of up to ~3 g/L/h and ~90% selectivity. Life cycle analysis confirmed a negative carbon footprint for the products. Unlike traditional production processes, which result in release of greenhouse gases, our process fixes carbon. These results show that engineered acetogens enable sustainable, high-efficiency, high-selectivity chemicals production. We expect that our approach can be readily adapted to a wide range of commodity chemicals.

59 BASIC BIOLOGICAL SCIENCES↗

Analysis of aircraft engine blade subject to ice impact

The ice impact on the engine blade made of layered composite is simulated. The ice piece is modeled as an equivalent spherical object and has the velocity opposite to that of the aircraft with direction parallel to the engine axis. Near the impact region and along the leading edge, the blade is assumed to be fully stressed and undergoes large deflection. A specified portion of the blade around the impact region is modeled. The effect of ice size and velocity on the average leading edge strain are investigated for a modified SR-2 model unswept composite propfan blade. Parametric studies are performed to study the response due to ice impact at various locations along the span. Also, the effects of engine speed on the strain and impact displacements are discussed. It is found that for a given engine speed, a critical ice speed exists that corresponds to the maximum strain and this critical speed increases with increase in the engine speed.

Reddy, E. S.↗

Analysis of aircraft engine blade subject to ice impact

The ice impact problem on an engine blade made of layered composite is simulated. The ice piece is modeled as an equivalent spherical object and has the velocity opposite to that of the aircraft with direction parallel to the engine axis. Near the impact region and along the leading edge, the blade is assumed to be fully stressed and undergoes large deflection. A specified portion of the blade around the impact region is modeled. The effect of ice size and velocity on the average leading edge strain are investigated for a modified SR-2 model unswept composite propfan blade. Parametric studies are performed to study the response due to ice impact at various locations along the span. Also, the effects of engine speed on the strain and impact displacements are discussed. It is found that for a given engine speed, a critical ice speed exists that corresponds to the maximum strain and this critical speed increases with increase in the engine speed.

Reddy, E. S.↗

raogroupuiuc/yl_variantcalling

NGS analysis for mutation analysis in Yarrowia lipolytica evolved strains. Yarrowia lipolytica, an oleaginous yeast, shows promise for industrial fermentation due to its robust acetyl-CoA flux and well-developed genetic engineering tools. However, its lack of an active xylose metabolism restricts the conversion of cellulosic sugars to valuable products. To address this, metabolic engineering, and adaptive laboratory evolution (ALE) were applied to the Y. lipolytica PO1f strain, resulting in an efficient xylose-assimilating strain (XEV). Whole-genome sequencing (WGS) of the XEV followed by reverse engineering revealed that the amplification of the heterologous oxidoreductase pathway and a mutation in the GTPase-activating protein gene (YALI0B12100g) might be the primary reasons for improved xylose assimilation in the XEV strain. When a sorghum hydrolysate was used, the XEV strain showed superior xylose consumption and lipid production compared to its parental strain (X123). This study advances our understanding of xylose metabolism in Y. lipolytica and proposes effective metabolic engineering strategies for optimizing lignocellulosic hydrolysates.

Deewan, Anshu↗

High strain rate deformation of aged TRIP Ti-10V-2Fe-3Al (wt.%) examined by in-situ synchrotron X-ray diffraction

Transformation Induced Plasticity (TRIP) is a promising avenue for tailoring the work hardening response of metastable $β$ titanium (Ti) alloys. Here we show that aged TRIP Ti-10V-2Fe-3Al (wt.%) maintains higher elongations and flow stresses as strain rate increases, if phase stability and microstructural characteristics are tuned. Low temperature aging influences the matrix $β$ phase stability by $ω$ phase precipitation, which affords a promising way to impact the TRIP effect and obtain desirable mechanical properties, ranging from high damping capacity to good strength/ductility combinations. Although TRIP is active during quasi-static and dynamic testing up to 2000 s –1 , increasing aging time and/or strain rate reduces the overall propensity for the TRIP effect and extent of transformation, which occurs rapidly just at the onset of yielding. Importantly, TRIP with $ω$ phase precipitation provides interesting alloying, microstructure, and property design strategies for engineering applications like lightweight protective structures, where high strains and the need for energy absorption are encountered.

36 MATERIALS SCIENCE↗