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At least 1,513 records · Page 84

Development of a Sustainable Green Chemistry Platform for Production of Acetone and Downstream Drop-in Fuel and Commodity Products Directly From Biomass Syngas Via a Novel Energy Conserving Route in Engineered Acetogenic Bacteria

LanzaTech and Oak Ridge National Laboratory have developed and scaled up a process for sustainable production of acetone and downstream drop-in fuel and commodity products directly from biomass syngas via a novel energy conserving route in engineered acetogenic bacteria. This process offers a safer and more environmentally-friendly production method for acetone production than the current phenol-dependent method, and the product has significantly lower GHG emissions. The developed process offers a cost competitive route to Acetone and enables biofuels at or below DOE’s $3 gge target. In addition, it also provides an attractive biological alternative to traditional sugar-based ABE fermentation, by enabling utilization of non-food biomass resources as fermentation feedstocks. Challenges overcome: • By-products formation: Byproduct 2,3-butandiol and 3-hydroxybutyrate reduce yield and stability. Addressed by elimination of both pathways in our chassis acetone production strain. • Cost competitiveness: Addressed by developing an integrated acetone strain and eliminating by-products to increase yield and stability. Optimized co-selectivity and coproductivity of acetone and ethanol instead of titer, based on technoeconomic analysis. • Limited enzyme variety: Addressed by genome mining of over 300 industrial strains in LanzaTech collection, identifying unique enzyme sequences, and refactoring these unique enzyme variants through LanzaTech’s engineering platform. Resulted in over 10x improvement in acetone production from gas. • Continuous process (stability): Identified bottleneck through detailed omics studies, addressed by integration of the acetone pathway on the chromosome. • Novel process to scale up: Addressed by demonstrating stable acetone production in a 80- L pilot reactor, with acetone productivity and selectivity comparable to those observed under the same conditions in 2-L reactors. We have demonstrated stable acetone production for over 7 days at commercial target rate and selectivity in 2-L reactors and have piloted the process.

09 BIOMASS FUELS↗

Development of a Sustainable Green Chemistry Platform for Production of Acetone and Downstream Drop-in Fuel and Commodity Products directly from Biomass Syngas via a Novel Energy Conserving Route in Engineered Acetogenic Bacteria

LanzaTech and Oak Ridge National Laboratory have developed and scaled up a process for sustainable production of acetone and downstream drop-in fuel and commodity products directly from biomass syngas via a novel energy conserving route in engineered acetogenic bacteria. This process offers a safer and more environmentally-friendly production method for acetone production than the current phenol-dependent method, and the product has significantly lower GHG emissions. The developed process offers a cost competitive route to Acetone and enables biofuels at or below DOE’s $3/gge target. In addition, it also provides an attractive biological alternative to traditional sugar-based ABE fermentation, by enabling utilization of non-food biomass resources as fermentation feedstocks. Challenges overcome: (1) By-products formation: Byproduct 2,3-butanediol and 3-hydroxybutyrate reduce yield and stability. Addressed by elimination of both pathways in the chassis acetone production strain. (2) Cost competitiveness: Addressed by developing an integrated acetone strain and eliminating by-products to increase yield and stability. Optimized co-selectivity and co-productivity of acetone and ethanol, based on technoeconomic analysis. (3) Limited enzyme variety: Addressed by genome mining of over 300 industrial strains in LanzaTech collection, identifying unique enzyme sequences, and refactoring these unique enzyme variants through LanzaTech’s engineering platform. Resulted in over 10x improvement in acetone production from gas. (4) Continuous process (stability): Identified bottleneck through detailed omics studies, addressed by integration of the acetone pathway on the chromosome. (5) Novel process to scale up: Addressed by demonstrating stable acetone production in an 80-L pilot reactor, with acetone productivity and selectivity comparable to those observed under the same conditions in 2-L reactors. LanzaTech has demonstrated stable acetone production for over 7 days at commercial target rate and selectivity in a 2-L reactor and have piloted the process.

09 BIOMASS FUELS↗

A mass spectrometry-based high-throughput screening method for engineering fatty acid synthases with improved production of medium-chain fatty acids

Microbial cell factories have been extensively engineered to produce free fatty acids (FFAs), key components of crucial nutrients, soaps, industrial chemicals, and fuels. Yet, our ability to control the specificity of microbially synthesized FFAs is still limited for medium-chain fatty acids (MCFAs), which is mainly due to lack of high-throughput approaches for FFA analysis. Here we report a mass spectrometry (MS)-based method for rapid profiling of MCFAs in Saccharomyces cerevisiae by using membrane lipids as a proxy. Specially, matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) MS was used to detect shorter acyl chain phosphatidylcholines from membrane lipids and the m/z peak ratio at 730 and 758 was used as an index for improved MCFA production. This yeast colony-based method can be performed at a rate of ~2 second per sample, which is much faster than the gold standard gas chromatography (GC)-MS (typically > 30 min per sample). To demonstrate the power of this method, we performed site-saturation mutagenesis of the yeast fatty acid synthase and identified 9 mutants that resulted in improved MCFA production relative to the wild-type strain. This colony-based MALDI-ToF MS screening method offers an alternative way for engineering microbial fatty acid compositions in a high-throughput manner.

59 BASIC BIOLOGICAL SCIENCES↗

Robust In-Situ Strain Measurements to Monitor CO 2 Storage

The goal of this project was to develop and demonstrate robust instrumentation to monitor the in-situ strain tensor in order to improve the reliability and security of CO 2 storage in geologic formations. We met the original goals of the project and the major overarching accomplishment is the advancement of strain tensor monitoring from an intriguing concept to a commercially available technology with a solid foundation of novel instruments supported by theoretical analyses and validation experiments. The main accomplishments of the project are summarized below. We designed, built and evaluated nine new optical fiber strainmeters and tiltmeters using Michelson interferometers to measure deformation with ultra-high resolution at both shallow and deep point locations in the subsurface. These are the robust strainmeters that motivated the title of the project. We designed, built and evaluated a novel method of measuring distributed strain in optical fibers with nanostrain resolution, and cm-scale location, and sampling into the seismic band. The new method is called Coherence-length-gated Microwave Photonics Interfereometry (CMPI). CMPI technology has advantages over existing commercial DAS and DSS methods. We developed and demonstrated capabilities to deploy instruments in the field and used them to measure strain caused by ambient signals like barometric pressure and tides, as well as induced signals like surface loading and pore pressure changes from pumping tests. We deployed a working strainmeter at 1,700 ft depth, slightly above an active reservoir. This is to our knowledge the greatest depth a strainmeter has been deployed and the techniques we used can readily be extended to greater depths. Optical fiber borehole tensor strainmeter techology was advanced from a TRL 4 at the start, to a TRL of 7 at the conclusion of the project. The project included advances in simulations and theoretical analyses. We developed and demonstrated a computational workflow that uses machine learning to reduce the computational requirements and make it practical to use Bayesian inversion to solve large numerical poroelastic analyses needed to interpret strain tensor field data. We evaluated the strain tensor fields and time series that would be caused by leaks of CO 2 or other fluids from reservoirs. These simulations demonstrated that signals from leaks could be measured with instruments developed for the project, opening a potentially new method for ensuring storage security. We showed that strains in caprock can be used to estimate pressure in a reservoir. This avoids the need to drill monitoring wells into the reservoir, and it expands the capabilities of monitoring in the caprock. The project includes a derivation and application of a novel analytical solution to the strains in the vicinity of a pressurized poroelastic inclusion. This solution explains field data measured during injeciton tests at the North Avant Field, and it will simplify future interpretation of strain tensor data. The project included a broad range of experiments, and of the most significant is the characterization of the strain tensor at an array three strainmeters during six injection tests at the North Avant Field, Oklahoma. This demonstrated repeatability of the strain signal measured by the new instruments developed for the project, and it showed similarities between the strain signal at shallow depths and pressure in the underlying reservoir. We also demonstrated that useful strain data can be measured at reservoir depths. This confirms that strain tensor data can be measured throughout the caprock over a reservoir. The project demonstrated the feasibility of using the strain tensor and distributed strain measured in caprock during a variety of different well tests where the pumping rate was constant, sinusoidal and positive, or a periodic square wave with zero net rate. This further strengthens the validity of using strain data to characterize reservoirs and aquifers. We also demonstrated that strain caused be fluctuations of air pressure and water pressure in the vadose zone can be measured and interpreted, suggesting that high resolution distributed strain measurements hold promise for monitoring the vadose zone. The project partially supported nine graduate students in the Environmental Engineering, Hydrogeology, Electrical Engineering programs at Clemson University. The research was described in nine journal papers, 23 talks and conference abstracts. Additional journal papers are in preparation. A new company called Tensora was started to provide strainmeter technology for commercial applications.

01 COAL, LIGNITE, AND PEAT↗

Metabolic engineering of the pentose phosphate pathway for enhanced limonene production in the cyanobacterium Synechocysti s sp. PCC 6803

Isoprenoids are diverse natural compounds, which have various applications as pharmaceuticals, fragrances, and solvents. The low yield of isoprenoids in plants makes them difcult for cost-efective production, and chemical synthesis of complex isoprenoids is impractical. Microbial production of isoprenoids has been considered as a promising approach to increase the yield. In this study, we engineered the model cyanobacterium Synechocystis sp. PCC 6803 for sustainable production of a commercially valuable isoprenoid, limonene. Limonene synthases from the plants Mentha spicata and Citrus limon were expressed in cyanobacteria for limonene production. Production of limonene was two-fold higher with limonene synthase from M. spicata than that from C. limon. To enhance isoprenoid production, computational strain design was conducted by applying the OptForce strain design algorithm on Synechocystis 6803. Based on the metabolic interventions suggested by this algorithm, genes (ribose 5-phosphate isomerase and ribulose 5-phosphate 3-epimerase) in the pentose phosphate pathway were overexpressed, and a geranyl diphosphate synthase from the plant Abies grandis was expressed to optimize the limonene biosynthetic pathway. The optimized strain produced 6.7mg/L of limonene, a 2.3-fold improvement in productivity. Thus, this study presents a feasible strategy to engineer cyanobacteria for photosynthetic production of isoprenoids.

59 BASIC BIOLOGICAL SCIENCES↗

Intensity measure adequacy assessment for nonlinear site response using Information Theory

Nonlinear site response modeling is a crucial aspect of Probabilistic Seismic Hazard Analysis. Site amplification models routinely rely on a rock intensity measure to characterize the strength of the bedrock motion. However, the adequacy of such intensity measures towards predicting amplifications across the oscillator period range has not been investigated in the literature. This paper analyzes the adequacy of rock intensity measures using state of the art criterion established in Performance-Based Earthquake Engineering and techniques from Information Theory. The efficiency and the sufficiency of several rock intensity measure are assessed. It was found that spectral accelerations at low periods usually are adequate for predicting amplifications across the period range. This supports the current practice of using Peak Ground Acceleration in Ground Motion Models. However, for extremely soft sites, which demonstrate nonlinear effects well into the period range, it is better practice to ensure that amplification and spectral acceleration share the same oscillator period. Lastly, for predicting the peak shear strain (an important parameter that controls nonlinearity of site response), Peak Ground Velocity is generally adequate, and this conclusion is in line with the definition of proxy shear strain.

42 ENGINEERING↗

Modular Engineering of Biomass Degradation Pathways

Production of fuels and chemicals from renewable lignocellulosic feedstocks is a promising alternative to petroleum-derived compounds. Due to the complexity of lignocellulosic feedstocks, microbial conversion of all potential substrates will require substantial metabolic engineering. Non-model microbes offer desirable physiological traits, but also increase the difficulty of heterologous pathway engineering and optimization. The development of modular design principles that allow metabolic pathways to be used in a variety of novel microbes with minimal strain-specific optimization will enable the rapid construction of microbes for commercial production of biofuels and bioproducts. In this review, we discuss variability of lignocellulosic feedstocks, pathways for catabolism of lignocellulose-derived compounds, challenges to heterologous engineering of catabolic pathways, and opportunities to apply modular pathway design. Implementation of these approaches will simplify the process of modifying non-model microbes to convert diverse lignocellulosic feedstocks.

09 BIOMASS FUELS↗

Synchrotron CT imaging of lattice structures with engineered defects

Understanding mechanical failure, crack propagation, and compressive behavior at the micrometer scale is essential for tailoring material properties for structural performance in cellular materials. Typically, modeling of traditional polymer foam materials is clouded by the lack of control in material morphology and its inherent stochastic structure. Additive manufacturing with sub-micrometer resolution provides a direct path for experimenters to specifically tailor structures needed by modelers to explicitly probe mechanical performance. Using laboratory-based 3D X-ray computed tomographic imaging (CT), the examination of deformation and damage provides a critical path to understand how these soft materials behave. Additionally, synchrotron CT yields realistic information at higher strain rates to directly validate the robustness of our finite element modeling. For this study, nanolithographic printing was employed to generate a series of engineered lattices with increasing levels of defects through the random removal of ligaments. These structures were mechanically tested and imaged with laboratory-based microCT. Additionally, synchrotron experiments were conducted in which the structures were imaged in 3D at 14 Hz during compression at a 0.4 s –1 strain rate. These 3D images show the changes in the structure as the ligaments bend, buckle and fracture in real time. This technique provides a robust framework for developing our methodologies and future exploration of engineered structures.

36 MATERIALS SCIENCE↗

The Transcriptome and Flux Profiling of Crabtree-Negative Hydroxy Acid-Producing Strains of Saccharomyces cerevisiae Reveals Changes in the Central Carbon Metabolism

Saccharomyces cerevisiae (S. cerevisiae) is the yeast cell factory of choice for the production of many biobased chemicals. However, it is a Crabtree-positive yeast and so shuttles a large portion of carbon into ethanol. Ethanol formation can be eliminated by deleting pyruvate decarboxylase (PDC) activity. It is not yet well understood how PDC-negative yeasts are affected when engineered to produce other products than ethanol. In this study, pathways are introduced for the production of three hydroxy acids (lactic, malic, or 3-hydroxypropionic acid [3HP]) into an evolved PDC-negative strain. These strains are characterized via transcriptome and flux profiling to elucidate the effects that the production of these hydroxy acids has on the host strain. Expression of lactic and malic acid biosynthesis pathways improved the maximum specific growth rate (μ max ) of the strain by 64% and 20%, respectively, presumably due to nicotinamide adenine dinucleotide regeneration. All strains show a very high flux (> 90% of glucose uptake) into the oxidative pentose phosphate pathway under batch fermentation conditions. The study, for the first time, directly compares the flux and transcriptome profiles of several hydroxy acid-producing strains of an evolved PDC-negative S. cerevisiae and suggests directions for future metabolic engineering.

13C-based metabolic flux analysis↗

Estimation of Errors in Stress Distributions Computed in Finite Element Simulations of Polycrystals

The accuracy of the stresses predicted from crystal plasticity-based finite element formulation depends on estimation and control of the errors associated with the discretization. In the current work, the errors in the stress distribution are estimated in virtual polycrystalline samples of α-phase titanium (hexagonal close-packed phase of Ti–6Al– 4 V). To estimate the error, the stress field, which does not possess inter-element continuity, is smoothed over a grain using an L 2 projection, thereby providing continuous stress distributions with inter-element continuity. The differences between the continuous (smooth) and discontinuous (raw) stress fields are calculated at individual Gauss quadrature points and used to estimate errors for corresponding elements and grains. Error estimations are performed for a Voronoi-tessellated microstructure, an equiaxed microstructure, and two microstructures with varying grain sizes for tensile loading extending into the fully plastic regime (≈ 5% extension). Magnitudes of the errors are found to depend on microstructural characteristics, particularly the shape and size of grains. Samples having variations in grain size or having less spherical grains exhibited higher errors than samples with uniformly sized, equiaxed grains, with the size variations having a more pronounced effect. In conclusion, errors correlate with proximity to grain boundaries at small (elastic) strains and with deformation-induced features (deformation bands) at large strains.

42 ENGINEERING↗

Ultrahigh Elastic Strain Energy Storage in Metal-Oxide-Infiltrated Patterned Hybrid Polymer Nanocomposites

Modulus of resilience, the measure of a material’s capacity to store and release elastic strain energy, is critical for realizing advanced mechanical actuation technologies in micro/nanoelectromechanical systems. In general, engineering the modulus of resilience is difficult because it requires asymmetrically increasing yield strength and Young’s modulus against their mutual scaling behavior. This task becomes further challenging if it needs to be carried out at the nanometer scale. Here, we demonstrate organic–inorganic hybrid composite nanopillars with one of the highest modulus of resilience per density by utilizing vapor-phase aluminum oxide infiltration in lithographically patterned negative photoresist SU-8. In situ nanomechanical measurements reveal a metal-like high yield strength (~500 MPa) with an unusually low, foam-like Young’s modulus (~7 GPa), a unique pairing that yields ultrahigh modulus of resilience, reaching up to ~24 MJ/m 3 as well as exceptional modulus of resilience per density of ~13.4 kJ/kg, surpassing those of most engineering materials. The hybrid polymer nanocomposite features lightweight, ultrahigh tunable modulus of resilience and versatile nanoscale lithographic patternability with potential for application as nanomechanical components which require ultrahigh mechanical resilience and strength.

36 MATERIALS SCIENCE↗

Thermally Grown Oxide Stress in PS-PVD and EB-PVD Thermal Barrier Coatings Observed at Various Lifetimes Via Synchrotron X-ray Diffraction

The current standard application method for thermal barrier coatings (TBCs) on turbine blades for jet engines is electron-beam physical vapor deposition (EB-PVD) due to its high strain tolerance and low thermal conductivity. An emerging deposition method, plasma-spray physical vapor deposition (PS-PVD), presents an opportunity for a tailorable microstructure, and non-line-of-sight deposition that is faster and less expensive. To compare the lifetime behavior of both PS-PVD and EB-PVD coatings, samples subjected to 300 and 600 thermal cycles were measured during a 1 h thermal cycle to determine the strains, which were converted to stress, in the thermally grown oxide (TGO) layer of the TBCs using synchrotron X-ray diffraction (XRD). Room temperature XRD measurements indicated among samples that PS-PVD coatings experienced greater variation in in-plane room temperature strain in the TGO after cycling than the EB-PVD coatings. In-situ XRD measurements indicated similar high-temperature strain and no spallation after 600 thermal cycles for both coatings. Further, microscopy imaging after cycling showed greater rumpling in PS-PVD coatings that led to different failure modes between the two coatings’ TGO layers. The tailorability of PS-PVD coatings allows for adjustments in the processing parameters to improve their overall performance after aging and bridge the differences between the two deposition methods.

36 MATERIALS SCIENCE↗

Parsed synthesis of pyocyanin via co-culture enables context-dependent intercellular redox communication

Abstract Background Microbial co-cultures and consortia are of interest in cell-based molecular production and even as “smart” therapeutics in that one can take advantage of division of labor and specialization to expand both the range of available functions and mechanisms for control. The development of tools that enable coordination and modulation of consortia will be crucial for future application of multi-population cultures. In particular, these systems would benefit from an expanded toolset that enables orthogonal inter-strain communication. Results We created a co-culture for the synthesis of a redox-active phenazine signaling molecule, pyocyanin (PYO), by dividing its synthesis into the generation of its intermediate, phenazine carboxylic acid (PCA) from the first strain, followed by consumption of PCA and generation of PYO in a second strain. Interestingly, both PCA and PYO can be used to actuate gene expression in cells engineered with the soxRS oxidative stress regulon, although importantly this signaling activity was found to depend on growth media. That is, like other signaling motifs in bacterial systems, the signaling activity is context dependent. We then used this co-culture’s phenazine signals in a tri-culture to modulate gene expression and production of three model products: quorum sensing molecule autoinducer-1 and two fluorescent marker proteins, eGFP and DsRed. We also showed how these redox-based signals could be intermingled with other quorum-sensing (QS) signals which are more commonly used in synthetic biology, to control complex behaviors. To provide control over product synthesis in the tri-cultures, we also showed how a QS-induced growth control module could guide metabolic flux in one population and at the same time guide overall tri-culture function. Specifically, we showed that phenazine signal recognition, enabled through the oxidative stress response regulon soxRS, was dependent on media composition such that signal propagation within our parsed synthetic system could guide different desired outcomes based on the prevailing environment. In doing so, we expanded the range of signaling molecules available for coordination and the modes by which they can be utilized to influence overall function of a multi-population culture. Conclusions Our results show that redox-based signaling can be intermingled with other quorum sensing signaling in ways that enable user-defined control of microbial consortia yielding various outcomes defined by culture medium. Further, we demonstrated the utility of our previously designed growth control module in influencing signal propagation and metabolic activity is unimpeded by orthogonal redox-based signaling. By exploring novel multi-modal strategies for guiding communication and consortia outcome, the concepts introduced here may prove to be useful for coordination of multiple populations within complex microbial systems.

Chun, Kayla↗

Mechanical Behavior of MinK and FiberFrax Board Insulation Materials Under Battery Packaging Relevant Conditions

We present a new collection of data on the load-stress relaxation-unload behavior of MinK and FiberFrax Board (FF) insulation materials used as pellets in-line with thermal battery electrochemical stacks. Both materials were subjected to standard thermal preparations, and then tested at room temperature. Intermediate term stress relaxation tests are presented (order 104 minutes of relaxation) showing that FF relaxation is not significantly stress or deformation dependent, but MinK is moderately so. Moreover, stress-strain curves associated with specimen unloading, reloading, and unloading again are presented for both materials. FF and MinK are substantially different here. Acute material variability is observed though test conditions and material preparations are standardized. A modeling approach is presented to empirically estimate the amount of stress relaxation at room temperature, and from this state, represent the unloading stress-strain behavior of both materials. This effort provides a complete framework for representing (in an engineering sense) both materials in thermal battery performance simulations.

36 MATERIALS SCIENCE↗

Kinetics of Failure in an Elastic Peridynamic Material

The dynamic behavior of an elastic peridynamic material with a nonconvex bond potential is studied. In spite of the material's inherently unstable nature, initial value problems can be solved using essentially the same techniques as with conventional materials. In a suitably constructed material model, small perturbations grow exponentially over time until the material fails. The time for this growth is computed explicitly for a stretching bar that passes from the stable to the unstable phase of the material model. This time to failure represents an incubation time for the nucleation of a crack. The finiteness of the failure time in effect creates a rate dependence in the failure properties of the material. Thus, the unstable nature of the elastic material leads to a rate effect even though it does not contain any terms that explicitly include a strain rate dependence.

36 MATERIALS SCIENCE↗

Verification of a Reduced-Order Extension Spring Model

Adopting reduced order models (ROMs) of springs lowers the computational cost of stronglink simulations. However, ROMs introduce currently unquantified error to such analyses. This study addresses that lack of data by comparing a hexahedral mesh to a commonly used ROM beam mesh. Two types of analyses were performed, a quasi-static displacement-controlled pull and a haversine shock, examining basic spring properties as well as dynamics and stress/strain data. Both tests showed good similarities between the hexahedral and beam meshes, especially when comparing reaction force and stress trends and maximums. Equivalent plastic strain results were not quite as favorable, indicating that the beam model may be less likely to correctly predict spring failure. Despite reducing computation times by over 48 hours in all shock cases, appropriate use of the ROM should carefully balance this advantage with its reduction in accuracy, especially when examining spring failure and outputting variables such as equivalent plastic strain.

42 ENGINEERING↗

(U) Blast Tube Rehabilitation (Final Design Review) [Slides]

Saddle design is being proposed to improve overall response of facility. No plasticity predicted in tube sections buying down risk of a catastrophic failure. Plasticity is predicted in the support for loadings representing 190# C4. The design has been iterated to decrease localized plasticity and buy down risk. It is recommended that no amount of explosive driver be placed in A36 sections. Previous work suggests yielding can develop in the tube section causing rupture. J2 colleagues have reviewed analysis results. Outstanding question of additional plastic strain is developing from subsequent uses.

42 ENGINEERING↗

Deriving Strain Modes From Vibrational Tests

Measurements and theoretical analysis complement each other. Experimental acceleration and strain data used to calculate coefficients of low-frequency vibrational modes of object under test. An iterative comparison of experimental and calculated strains give modal model of improved accuracy that predicts strains under operating conditions. Method useful in fatigue life and reliability analyses of buildings, pumps, engines, vehicles, and other systems subject to vibrations and loud noises during operation.

Young, J. W.↗