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At least 937 records · Page 52

Parametric Failures in COTS Capacitors

Insertion of COTS components into hi-rel systems require extensive environmental testing of the parts that often results in parametric failures. Based on experience with commercial and hi-rel PME and BME ceramic capacitors and polymer and MnO2 cathode tantalum capacitors, this presentation discusses two major reasons for parametric failures. One is due to the marketing pressure that forces manufacturers to squeeze performance of COTS components thus leaving insufficient margin between the rated and actual characteristics. Another, and probably a more serious reason is degradation of characteristics caused by physico-chemical processes in materials under environmental stresses. Reliability assessment requires development of models to predict behavior of the parts during applications based on accelerated testing that might be outside the specified conditions and target applications. Examples of models developed to simulate and predict wear-out failures are presented. Risks associated with overstressing during HALT are discussed.

Teverovsky, Alexander↗

Displacement Damage Induced Dark Current and Random Telegraph Signal in P-Type Deep-Trench Pinned Photogates

This study focuses on displacement damage induced dark current and Random Telegraph Signal (RTS) in STMicroelectronics P-Type photogate, a device moving away from the 4T Pinned-Photodiode, with a sensitive volume exclusively made of boron-doped P-type epitaxy, and collecting holes instead of electrons. Results show a typical trend for the shape of dark current and RTS degradation in CMOS Image Sensors with the exception that the measured generation rates seem 2 times smaller than typical literature values in irradiated silicon. Different hypothesis on the origin of this discrepancy are explored.

displacement damage dose↗

Thermal Oxo-degradation and Catalytic Upgrading of Plastic Waste to Light Olefins for a Circular Economy

The commercialization of conventional pyrolysis of plastic in an inert atmosphere has been hindered by large thermal requirements stemming from long reaction rates. The rate of thermal depolymerization of waste plastics can be accelerated by the addition of oxygen in a process known as thermal oxo-degradation (TOD). This study offers the prospect of TOD to upcycle postconsumer waste rapidly and efficiently. Using moderate temperatures and small amounts of air in a fluidized bed reactor, we demonstrated that waste high-density polyethylene and polypropylene are rapidly deconstructed to condensable products. These condensable products were catalytically upgraded in a micropyrolysis reactor using commercially available zeolite (HZSM-5) to monomeric olefins. The olefin yields proved to be greater than those achieved through the catalytic upgrading of condensable products from (nonoxidative) the pyrolysis of the same plastic wastes. The coupling of TOD with catalytic upgrading proves to be an energy-efficient pathway in a plastics circular economy for the production of light olefins from wastes.

Enthalpy↗

Mn(III)-mediated bisphenol a degradation: Mechanisms and products

Bisphenol A (BPA) is a high production volume chemical with potential estrogenic effects susceptible to abiotic degradation by MnO 2 . BPA transformation products and reaction mechanisms with MnO 2 have been investigated, but detailed process understanding of Mn(III)-mediated degradation has not been attained. Rapid consumption of BPA occurred in batch reaction vessels with 1 mM Mn(III) and 63.9 ± 0.7% of 1.76 ± 0.02 μmol BPA was degraded in 1 hour at circumneutral pH. BPA was consumed at 1.86 ± 0.09-fold higher rates in vessels with synthetic MnO 2 comprising approximately 13 mol% surface-associated Mn(III) versus surface-Mn(III)-free MnO 2 , and 10–35% of BPA transformation could be attributed to Mn(III) during the initial 10-min reaction phase. High-resolution tandem mass spectrometry (HRMS/MS) analysis detected eight transformation intermediates in reactions with Mn(III), and quantum calculations proposed 14 BPA degradation products, nine of which had not been observed during MnO 2 -mediated BPA degradation, suggesting mechanistic differences between Mn(III)- versus MnO 2 -mediated BPA degradation. Finally, the findings demonstrate that both Mn(III) and Mn(IV) can effectively degrade BPA and indicate that surface-associated Mn(III) increases the reactivity of synthetic MnO 2 , offering opportunities for engineering more reactive oxidized Mn species for BPA removal.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Pharmacological Inhibitors of the Proteosome in Atrophying Muscles

It is now clear that the marked loss of muscle mass that occurs with disuse, denervation or in many systemic diseases (cancer cachexia, sepsis, acidosis, various endocrine disorders) is due primarily to accelerated degradation of muscle proteins, especially myofibrillar components. Recent work primarily in Dr. Goldberg's laboratory had suggested that in these diverse conditions, the enhancement of muscle proteolysis results mainly from activation of the Ub-proteasome degradative pathway. In various experimental models of atrophy, rat muscles show a common series of changes indicative of activation of this pathway, including increases in MRNA for Ub and proteasome subunits, content of ubiquitinated proteins, and sensitivity to inhibitors of the proteasome. In order to understand the muscle atrophy seen in weightlessness, Dr. Goldberg's laboratory is collaborating with Dr. Baldwin in studies to define the changes in these parameters upon hind-limb suspension. Related experiments will explore the effects on this degradative system of exercise regimens and also of glucocorticoids, which are known to rise in space personnel and to promote muscle, especially in inactive muscles. The main goals will be: (A) to define the enzymatic changes leading to enhanced activity of the Ub-proteasome pathway in inactive muscles upon hind-limb suspension, and the effects on this system of exposure to glucocorticoids or exercise; and (B) to learn whether inhibitors of the Ub-proteasome pathway may be useful in retarding the excessive proteolysis in atrophying muscles. Using muscle extracts, Dr. Goldberg's group hopes to define the rate-limiting, enzymatic changes that lead to the accelerated Ub-conjugation and protein degradation. They have recently developed cell-free preparations from atrophying rat muscles, in which Ub-conjugation to muscle proteins is increased above control levels. Because these new preparations seem to reproduce the changes occurring in vivo, they will analyze in depth extracts from normal and atrophying muscles to compare the activities of the Ub-activating enzyme (El), the various LTh-carrier proteins (E2s), and Ub-protein ligases (E3s). Recent studies of other types of muscle wasting -suggest a very important role in muscle proteolysis of certain ubiquitination enzymes, E214k and E3-alpha(i.e. components of the "N-end pathway"). Future studies will focus in understanding their role and test whether they are in fact critical for muscle atrophy in vivo. Since weightlessness leads to a specific loss of contractile proteins and to a switching of myosin isotypes, Dr. Goldberg's group will attempt to identify the ubiquitination enzymes specifically involved in myosin degradation both in normal muscle and after hind-limb suspension.

Goldberg, Alfred↗

Coupled Biotic-Abiotic Processes Control Biogeochemical Cycling of Dissolved Organic Matter in the Columbia River Hyporheic Zone

A critical component of assessing the impacts of climate change on watershed ecosystems involves understanding the role that dissolved organic matter (DOM) plays in driving whole ecosystem metabolism. The hyporheic zone—a biogeochemical control point where ground water and river water mix—is characterized by high DOM turnover and microbial activity and is responsible for a large fraction of lotic respiration. Yet, the dynamic nature of this ecotone provides a challenging but important environment to parse out different DOM influences on watershed function and net carbon and nutrient fluxes. We used high-resolution Fourier-transform ion cyclotron resonance mass spectrometry to provide a detailed molecular characterization of DOM and its transformation pathways in the Columbia river watershed. Samples were collected from ground water (adjacent unconfined aquifer underlying the Hanford 300 Area), Columbia river water, and its hyporheic zone. The hyporheic zone was sampled at five locations to capture spatial heterogeneity within the hyporheic zone. Our results revealed that abiotic transformation pathways (e.g., carboxylation), potentially driven by abiotic factors such as sunlight, in both the ground water and river water are likely influencing DOM availability to the hyporheic zone, which could then be coupled with biotic processes for enhanced microbial activity. The ground water profile revealed high rates of N and S transformations via abiotic reactions. The river profile showed enhanced abiotic photodegradation of lignin-like molecules that subsequently entered the hyporheic zone as low molecular weight, more degraded compounds. While the compounds in river water were in part bio-unavailable, some were further shown to increase rates of microbial respiration. Together, river water and ground water enhance microbial activity within the hyporheic zone, regardless of river stage, as shown by elevated putative amino-acid transformations and the abundance of amino-sugar and protein-like compounds. This enhanced microbial activity is further dependent on the composition of ground water and river water inputs. Our results further suggest that abiotic controls on DOM should be incorporated into predictive modeling for understanding watershed dynamics, especially as climate variability and land use could affect light exposure and changes to ground water essential elements, both shown to impact the Columbia river hyporheic zone.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Degradation and ESR Failures in MnO2 Chip Tantalum Capacitors

Equivalent series resistance (ESR) of chip tantalum capacitors determines the rate of energy delivery and power dissipation thus affecting temperature and reliability of the parts. Employment of advanced capacitors with reduced ESR decreases power losses and improves efficiency in power systems. Stability of ESR is essential for correct operations of power units and might cause malfunctioning and failures when ESR becomes too high or too low. Several cases with ESR values in CWR29 capacitors exceeding the specified limit that were observed recently raised concerns regarding environmental factors affecting ESR and the adequacy of the existing screening and qualification testing. In this work, results of stress testing of various types of military and commercial capacitors obtained over years by GSFC test lab and NEPP projects that involved ESR measurements are described. Environmental stress tests include testing in humidity and vacuum chambers, temperature cycling, long-term storage at high temperatures, and various soldering simulation tests. Note that in many cases parts failed due to excessive leakage currents or reduced breakdown voltages. However, only ESR-related degradation and failures are discussed. Mechanisms of moisture effect are discussed and recommendations to improve screening and qualification system are suggested.

Equivalent Series Resistance (ESR)↗

Crack Growth Rate and Fracture Toughness of Stainless Steel Welds in Light Water Reactor Environment

Austenitic stainless steel welds play a crucial role in maintaining the structural integrity of light water reactors. Within the reactor core, stainless steel welds endure a harsh service environment including elevated temperature, corrosive coolant, and intense neutron irradiation. Long-term exposure to such a severe service environment could lead to performance degradation. To ensure the safe and economic operation of light water reactors, this study performed crack growth rate and fracture toughness tests on irradiated and thermally aged weld samples harvested from decommissioned reactor components. In a low-corrosion-potential environment, stainless steel welds showed resistance to stress corrosion cracking, and the measured crack propagation rates were comparable to those of wrought stainless steels under similar test conditions. However, a marked decline in fracture toughness was observed in weld samples, particularly in irradiated samples. Fracture instability was observed at ~0.8 dpa, significantly lower than those observed in wrought stainless steels. Further research at higher doses is needed to evaluate the dose dependence of irradiation-induced embrittlement, and to understand the interplay between thermal aging and neutron irradiation in stainless steel welds.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Linking microstructure to creep behavior in vertically and horizontally built LPBF Haynes 282 compared with wrought material via θ -projection

Laser Powder Bed Fusion (LPBF) has emerged as a promising route for fabricating intricate geometries in high-performance alloys. Haynes 282 (H282) is a strong candidate for applications such as heat exchangers or engines due to its excellent creep strength and thermal stability; however, the long-term creep behavior of LPBF-processed H282 remains poorly understood. In this study, the θ -projection method is used to analyze and extrapolate the creep behavior of vertically built LPBF, horizontally built LPBF, compared to wrought H282 tested at 816 °C. Vertically built LPBF H282 exhibits the lowest minimum creep rate (MCR), while the horizontally built condition shows a higher MCR comparable to that of wrought H282. Despite these differences, both LPBF conditions exhibit significantly shorter rupture life and reduced rupture strain than the wrought material, with the most severe degradation observed in the horizontal builds, consistent with an earlier onset of tertiary creep and accelerated strain-rate evolution. Microstructural characterization reveals that both LPBF and wrought H282 exhibit abundant twin-related boundary character; however, their grain boundary topologies differ markedly. The wrought alloy contains a higher fraction of low-angle grain boundaries and continuous twin lamellae, whereas the LPBF microstructure is characterized by a suppressed low-angle boundary population and fragmented twin-related boundaries embedded within irregular high-angle grain boundary networks. Fractographic analysis further reveals predominantly intergranular cracking in LPBF H282, accompanied by grain-boundary-decorated carbides, Al 2 O 3 inclusions, and high-aspect-ratio pores. These results demonstrate that grain boundary topology, rather than minimum creep rate alone, plays a critical role in governing creep damage accumulation and rupture behavior in LPBF and wrought H282.

Creep↗

Enabling Long Term, Shelf‐Stable Perovskite Solar Cells: Alumina Barriers to Protect Perovskite Devices from Moisture and Oxygen

Current encapsulation architectures for thin-film metal halide perovskite do not adequately eliminate all ambient stressors. Here, we develop low transmission rate barrier layers using atomic layer deposited (ALD) aluminum oxide grown directly on the completed photovoltaic (PV) device stack to provide an additional seal, prior to full packaging. We investigate the effect of deposition temperature and oxidant chemistries on the barrier growth for protection of perovskite photovoltaic devices. We characterize the layers individually, then integrate into devices to detail the tradeoff between protection and deposition compatibility. To enhance compatibility and impermeability, we present an approach using water as the aluminum oxidant during nucleation and switching from water to ozone for the remainder of the growth. At 50 nm, the barrier results in a water vapor transmission rate (WVTR) of 4.5·10 −4 g/m 2 /day, and 1,000-hour device stability under 45°C with 85% relative humidity without further packaging. This barrier provides sufficient protection to enable minimal degradation of the perovskite solar cells while completely submerged in water for 140 minutes. Additionally, we characterize the oxygen transmission rate (OTR) to be 0.49 cm 3 /m 2 /day at 23°C and 0% relative humidity, which is 1.5 orders of magnitude improvement over the OTR of the current encapsulation.

14 SOLAR ENERGY↗

In Situ Reliability Investigation of All-Vanadium Redox Flow Batteries by a Stable Reference Electrode

Redox flow batteries (RFBs) have been studied over the past several decades as a promising candidate for stationary energy storage applications. It is therefore important to understand the reliability of RFBs and the mechanisms that cause degradation with time. Contributions from individual electrodes (cathode or anode) are difficult to separate especially for long-term cycle testing, due to the lack of a stable reference electrode. In our work, the reliability and degradation mechanisms of an all-vanadium RFB were investigated by a stable reference electrode based on the dynamic hydrogen electrode (DHE). With factor exploration and design optimization, the newly developed DHE reference electrode demonstrated high accuracy and long-term stability that enables in-situ monitoring of individual electrode signals (potentials and impedances) over hundreds of cycles in a vanadium RFB. This approach enables the full cell degradation to be separated and the degradation attributed to contributions from the cathode and anode. The cathode and anode were found to play quite different roles in the increase in overpotential of the vanadium RFB during long-term cycling. The anode reaction limited both the charge and discharge capacity over 100 cycles. In addition, the negative side also appeared to be the rate limiting factor throughout cycling as determined by EIS measurement. The cathode contributed to the performance degradation as cycling exceeded 50 cycles.

Huang, Qian↗

Versatile microbial communities rapidly assimilate ammonium hydroxide-treated plastic waste

Abstract Waste plastic presently accumulates in landfills or the environment. While natural microbial metabolisms can degrade plastic polymers, biodegradation of plastic is very slow. This study demonstrates that chemical deconstruction of polyethylene terephthalate (PET) with ammonium hydroxide can replace the rate limiting step (depolymerization) and by producing plastic-derived terephthalic acid and terephthalic acid monoamide. The deconstructed PET (DCPET) is neutralized with phosphoric acid prior to bioprocessing, resulting in a product containing biologically accessible nitrogen and phosphorus from the process reactants. Three microbial consortia obtained from compost and sediment degraded DCPET in ultrapure water and scavenged river water without addition of nutrients. No statistically significant difference was observed in growth rate compared to communities grown on DCPET in minimal culture medium. The consortia were dominated by Rhodococcus spp., Hydrogenophaga spp., and many lower abundance genera. All taxa were related to species known to degrade aromatic compounds. Microbial consortia are known to confer flexibility in processing diverse substrates. To highlight this, we also demonstrate that two microbial consortia can grow on similarly deconstructed polyesters, polyamides, and polyurethanes in water instead of medium. Our findings suggest that microbial communities may enable flexible bioprocessing of mixed plastic wastes when coupled with chemical deconstruction.

59 BASIC BIOLOGICAL SCIENCES↗

Effects of hydraulic fluid CVCM on silver-Teflon

Changes in the solar absorptance of silver-Teflon (Ag/FEP) surfaces were measured after contamination by the outgassing products of hydraulic fluid and irradiation by solar simulators. Although radiator surfaces are selected for stability to the solar ultraviolet and particulate environment, contaminant deposition and their interaction with solar exposure can degrade their performance. The test procedure to deposit the hydraulic fluid collected volatile condensible material (CVCM), irradiate, and determine solar absorptance changes by reflectance measurements using a spectrophotometer and a solar simulator is described. The tests showed that for the estimated deposition rates on the Ag/FEP, the hydraulic fluid CVCM fraction remaining under normal conditions (above -40 C) would not degrade its performance. The residual CVCM can be easily removed from Ag/FEP by wiping with a solvent saturated flannel. The changes in solar absorptance with the CVCM film thickness and the radiation exposure duration are reported, concluding that vacuum ultraviolet radiation did not prevent easy removal of CVCM.

Muscari, J. A.↗

Evaluation of Oxygen Consumption as a Sensitive Measure of Electrical Cable Polymer Insulation Degradation

Polymers used as dielectric materials in nuclear grade electrical cables, such as cross-linked polyethylene (XLPE) and ethylene-propylene rubber (EPR), are long-lived and durable. Estimating the decades-long useful service life of polymers experimentally in a feasible timeframe of weeks or months requires greatly accelerated aging in the laboratory at elevated temperatures and/or gamma dose rates. Many limitations of accelerated aging, such as due to artefacts of the process including diffusion limited oxidation and dose rate effects, have been identified that can make accurate lifetime prediction at service conditions challenging. This can be due to differences in degradation mechanisms between rapid and extended aging or to the presence of induction periods during which early aging is undetectable using common metrics of polymer insulation characterization such as tensile elongation at break. Conceptually, an ideal measure of aging would track clearly and predictably with material changes throughout the lifecycle of the evolving material state from early aging through end of useful life. With such a metric accelerated aging could be performed at much milder conditions, conditions closer to those found in long term service, and end of life calculated from the early data. Oxygen consumption has previously been proposed as a sensitive measure of insulation polymer oxidation that might meet such desirable characteristics. In this work, nuclear cable insulation samples of the two most common types, XLPE and EPR, from two of the most sourced manufacturers, RSCC and The Okonite Company, were subjected to gamma irradiation at dose rates of 100, 200, and 300 Gy/hr for up to 42 days at 26°C. The aged samples were used to evaluate oxygen consumption as a sensitive measure of aging for these materials and these conditions. For comparison and validation, the aged samples were also characterized using the more conventional tests of mass change, density, carbonyl index, and yellowness index. For both materials, oxygen consumption during irradiation was found to track consistently with applied dose—oxygen consumption rate was linear with time. Mass change and carbonyl index were also clearly proportional to exposure dose. Yellowness index, a surface sensitive measure, was observed to increase linearly with dose at low exposure levels before plateauing. Sample density was not observed to change under the conditions explored. Oxygen consumption was found to be a particularly sensitive metric to measure the degradation of polymeric cable insulation exposed to radiation. Oxygen consumption was compared to Arrhenius and time-temperature-superposition methods for activation energy calculation using literature date and found to be a useful and viable alternative for lifetime prediction. The techniques developed in this work are anticipated to be valuable in the estimation of insulation aging rates under mild conditions approaching service conditions. Direction comparison of useful life prediction by oxygen consumption with prediction via Arrhenius methodology for thermal aging and existing methods for radiation and combined thermal/radiation aging would serve to further validate this promising approach and refine our understanding of material-specific aging rates.

36 MATERIALS SCIENCE↗

Elastomeric Seal Performance after Terrestrial Ultraviolet Radiation Exposure

Ultraviolet radiation was evaluated to determine its negative effects on the performance of elastomeric gas pressure seals. The leak rates of the silicone elastomer S0383-70 O-ring test articles were used to quantify the degradation of the seals after exposure to vacuum-ultraviolet and/or middle-to-near-ultraviolet wavelength radiation. Three groups of seals were exposed in terrestrial facilities to 115-165 nm wavelength radiation, 230-500 nm wavelength radiation, or both spectrums, for an orbital spaceflight equivalent of 125 hours. The leak rates of the silicone elastomer S0383-70 seals were quantified and compared to samples that received no radiation. Each lot contained six samples and statistical t-tests were used to determine the separate and combined influences of exposure to the two wavelength ranges. A comparison of the mean leak rates of samples exposed to 115-165 nm wavelength radiation to the control specimens showed no difference, suggesting that spectrum was not damaging. The 230-500 nm wavelength appeared to be damaging, as the mean leak rates of the specimens exposed to that range of wavelengths, and those exposed to the combined 115-165 nm and 230-500 nm spectrums, were significantly different from the leak rates of the control specimens. Most importantly, the test articles exposed to both wavelength spectrums exhibited mean leak rates two orders of magnitude larger than any other exposed specimens, which suggested that both wavelength spectrums are important when simulating the orbital environment.

leakage↗

Exploration of Failure and Potential Damage Markers in Ti-6Al-4V

To improve prognostics for predicting component life, coupon tests were conducted on Ti-6Al-4V to investigate the development of damage. Various test types were employed (tension, creep, stress relaxation, and fatigue) under a wide range of temperatures and loading rates to engage various amounts of time-dependent and damage behavior. Both stiffness degradation and Poisson’s ratio were monitored to see if they were useful in indicating the initiation and accumulation of damage and subsequently the onset of failure. Stiffness exhibited large decreases with increasing strain when calculated using engineering stress and strain. However, minimal stiffness change was observed when using the true-stress and true- strain values. Detailed microscopy and image analysis were performed to document the actual physical damage that occurred within the various specimens. Two types of damage were observed: shallow surface cracks and internal pores. The severity of damage was greater in the region of localized necking and diminished further away from the neck. The area fraction of pores was higher in the necked region, yet even there it did not exceed more than a few percent and, in most cases, was significantly lower. It was observed that conditions that induced more time-dependent deformation (e.g., slower strain rate testing) resulted in a larger number of pores and larger pore sizes. The pores were always found to occur in the softer β-phase. The severity of observed damage was inconsistent with the large drop in engineering stiffness, but was more aligned with stiffness changes from the true-stress and -strain values. Poisson’s ratio was monitored on every test and showed promise as a damage indicator. It was found to be more sensitive than either the ultimate tensile strength or the inception of tertiary creep. However, calculation of Poisson’s ratio was found to be more involved than expected because of anisotropy and the presence of strain gradients along the specimen at large applied strains. First, the material had a moderate texture and resulted in anisotropy of the strains. Second, the evolution of Poisson’s ratio as a function of axial strain showed strange and unexpected behaviors, which necessitated more extensive analyses of its development. This led to an investigation on the use of small versus large strain approaches and the importance of making consistent measurements; that is, axial and transverse strain measurements over the same volume of material. Additionally, finite element analysis of a tensile sample indicated that non- uniform strain occurred in the sample prior to attaining the maximum strength. This, along with the location at which transverse strain was measured, had a significant influence on the resulting values of Poisson’s ratio. Based on this, lessons learned are offered for future testing.

Ti-6Al-4V↗

Interleaver design for trellis-coded differential 8-PSK modulation with non-coherent detection

The effect of finite interleaver size on bit error rate (BER) performance of coded 8-DPSK is determined by means of computer simulations. The losses evaluated in this way include the SNR degradation due to the timing and frequency errors of the symbol synchronizer and the automatic frequency control (AFC) of the receiver. BER measurements are presented using a conventional 2/3 rate convolutional 8-state trellis-code for typical Rayleigh and Rician fading channels. It is shown that for a Rician channel with a Rician parameter of 7 dB, a Doppler spread of 100 Hz and a data rate of 2400 bps, an interleaver with size 16 x 16 symbols performs nearly as well as a very large interleaver. It is also shown that for very fast Rayleigh channels, the BER-curves flatten out at large SNR.

Edbauer, Franz↗

Systems Engineering of Rhodococcus opacus to Enable Production of Drop-in Fuels from Lignocellulose

Production of drop-in fuels from lignocellulose using Rhodococcus opacus PD630 (hereafter R. opacus) is a challenging goal. During the grant period we have pushed the field forward significantly in several areas of research. Towards the end goal of accelerating the adoption of R. opacus in biofuel production, during the grant period we have expanded the phenotypic characterization of R. opacus grown in single aromatic (model lignocellulosic) compounds or their mixtures, modeling the growth conditions in lignocellulosic biomass. Harnessing the power of adaptive evolution, we produced evolved R. opacus isolates with superior lignin valorization capabilities and identified differentially expressed genes and pathways after adaptation. We used next generation multi-omic techniques such as genomic, transcriptomic, and metabolomic analyses, to identify the catabolic pathways used by R. opacus to degrade aromatic compounds and funnel these degradation products into central metabolism, as well as the aromatic transport genes required for increased tolerance and utilization. Taking this information one step further, we identified endogenous transcription factors and regulatory mechanisms important for degradation of five model aromatic compounds. To accurately estimate R. opacus growth and consumption on model lignin compounds we pioneered the use of novel extraction procedures prior to GC-MS analysis. Alongside 13 C-metabolic flux analysis, we have elucidated the metabolic routes preferred by Rhodococcus opacus during aromatic compound degradation. Finally, we used in tandem lipidomics and high-resolution mass spectrometry to identify the modulation of mycolic acids and phospholipid membrane composition modification as a strategy for aromatic tolerance in R. opacus. Being a non-model organism, R. opacus lacks the breadth of tools and technical foundation which drive biofuel research in more well-understood microbes such as Escherichia coli. To reduce this burden for use, we designed and produced new tools for genomic manipulation and engineering in R. opacus. These engineering breakthroughs support efficient genomic editing, enabling gene overexpression, repression, and genetic alteration. Using these tools, we have generated synthetically engineered strains with increased lipogenesis and growth, both positive traits required for increased lignin valorization. Optimizing engineered strains for biofuel production from lignocellulose requires extremely sophisticated synthetic rewiring of metabolism. To facilitate systems-level reorganization of metabolism in R. opacus, we created a genome-scale model that accurately predicts metabolic flux and growth rates on the aromatic compound phenol. Lignin requires extensive pre-treatment before biological degradation by R. opacus. Towards an eventual goal of degrading real-world lignin, we developed new depolymerization processes to generate lignin breakdown products (LBP). We optimized LBP storage and composition analysis techniques, enabling accurate prediction of specific LBP compound integration into cell wall components. Overall, through the work funded by this grant we generated 20 manuscripts (17 published, 3 in review/preparation), methods for increased accuracy in metabolomics of aromatic compounds, multiple genetic tools for altering the R. opacus genome, genome scale models for predicting flux through metabolic pathways, as well as multi-omic data for community use. The work funded by this grant has increased the knowledge of aromatic degradation in bacteria and advanced our efforts to optimize R. opacus for lignin valorization.

09 BIOMASS FUELS↗