Engineering Papers⌕ Search

SEARCH · Engineering Papers

Results for “Strain improvement”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 667 records · Page 37

ACT University Collaboration Proposal: Role of manufacturing defects on material failure under dynamic loading for developing enhanced failure models and theories (Final Report)

Research on the tensile behavior of additively manufactured 316L stainless steel coupon specimens at increasing strain rates was conducted over the past year at Penn State. Dynamic loading rates into the 1000 1/s loading rates were performed on specimens with nearly 1.32” gage lengths. Stress-strain plots show ductile and plastic behavior well beyond 12% strain with necked specimens, having smaller effective gage lengths showing up to 56% ultimate strain. Additional tests performed on compact tension specimens helped with simulation work to understand the deformation behavior. Using finite elements, it was possible to determine the feasibility of a comprehensive experimental validation study towards an improved model for AM material failure – specifically, the Bai-Wierzbicki approach which accounts for lode angle and triaxiality. A non-significant number of tests are projected for eight specific failure nodes, with additional replicates to provide strain-rate capabilities to the existing formulation. One such correction factor is explored for quasi-static, notched specimens. Finally, stress intensity factor was explored using a set of compact-tension specimens which would also provide useful validation data for any simulation work.

36 MATERIALS SCIENCE↗

Laminate behavior for SiC fiber-reinforced reaction-bonded silicon nitride matrix composites

The room temperature mechanical properties of SiC fiber reinforced reaction-bonded silicon nitride matrix composite laminates (SiC/RBSN) have been measured. The laminates contained approx 30 volume fraction of aligned 142-micron diameter SiC fiber in a porous RBSN matrix. Three types of laminate studied were unidirectional: (1) (0) sub 8, (2) (10) sub 8, and (3) (45) sub 8, and (90) sub 8; cross plied laminates (0 sub 2/90 sub 2); and angle plied laminates: (+45 sub 2/-45 sub 2). Each laminate contained eight fiber plies. Results of the unidirectionally reinforced composites tested at various angles to the reinforcement direction indicate large anisotropy in in-plane properties. In addition, strength properties of these composites along the fiber direction were independent of specimen gage length and were unaffected by notches normal to the fiber direction. Splitting parallel to the fiber at the notch tip appears to be the dominant crack blunting mechanism responsible for notch insensitive behavior of these composites. In-plane properties of the composites can be improved by 2-D laminate construction. Mechanical property results for (0 sub 2/90 sub 2)sub s and (+45/-45 sub 2) sub s laminates showed that their matrix failure strains were similar to that for (0) sub 8 laminates, but their primary elastic moduli, matrix cracking strengths, and ultimate composite strengths were lower. The elastic properties of unidirectional, cross-ply, and angle-ply composites can be predicted from modified constitutive equations and laminate theory. Further improvements in laminate properties may be achieved by reducing the matrix porosity and by optimizing the bond strength between the SiC fiber and RBSN matrix.

Rhatt, R. T.↗

Laminate behavior for SiC fiber-reinforced reaction-bonded silicon nitride matrix composites

The room temperature mechanical properties of SiC fiber reinforced reaction-bonded silicon nitride matrix composite laminates (SiC/RBSN) have been measured. The laminates contained approx 30 volume fraction of aligned 142-micron diameter SiC fiber in a porous RBSN matrix. Three types of laminate studied were unidirectional: (1) (0) sub 8, (2) (10) sub 8, and (3) (45) sub 8, and (90) sub 8; cross plied laminates (0 sub 2/90 sub 2); and angle plied laminates: (+45 sub 2/-45 sub 2). Each laminate contained eight fiber plies. Results of the unidirectionally reinforced composites tested at various angles to the reinforcement direction indicate large anisotropy in in-plane properties. In addition, strength properties of these composites along the fiber direction were independent of specimen gage length and were unaffected by notches normal to the fiber direction. Splitting parallel to the fiber at the notch tip appears to be the dominant crack blunting mechanism responsible for notch insensitive behavior of these composites. In-plane properties of the composites can be improved by 2-D laminate construction. Mechanical property results for (0 sub 2/90 sub 2) sub s and (+45/-45 sub 2) sub s laminates showed that their matrix failure strains were similar to that for (0) sub 8 laminates, but their primary elastic moduli, matrix cracking strengths, and ultimate composite strengths were lower. The elastic properties of unidirectional, cross-ply, and angle-ply composites can be predicted from modified constitutive equations and laminate theory. Further improvements in laminate properties may be achieved by reducing the matrix porosity and by optimizing the bond strength between the SiC fiber and RBSN matrix.

Bhatt, Ramakrishna T.↗

Probiotics in the Space Food System: Delivery, Microgravity Effects, and the Potential Benefit to Crew Health

As mission distance and duration increase, the need grows for non‐invasive disease prevention and immunomodulation, especially given the limited medical response capability expected for these missions and the immune dysregulation documented in crew. Additionally, changes in diet, lifestyle, antibiotic usage, and the environmental stresses during spaceflight may alter crewmembers' intestinal microbiome. The addition of probiotic bacteria to the space food system is expected to confer immunostimulatory benefits on crewmembers, with the potential to counteract the immune dysregulation that has been documented in spaceflight. Based on previous studies that demonstrated unique microbiological responses to the low shear environment of spaceflight, probiotic organisms hold the potential to induce enhanced beneficial responses through mechanisms, such as beneficial interactions with human immune cells and repression of colonization of pathogens on the mucosa. The work presented here will begin to address two research gaps related to providing probiotics in spaceflight: 1) delivery, and 2) the effect of the low shear microgravity environment on probiotic attributes. The probiotic Lactobacillus acidophilus was selected for investigation due to its wide commercial use and documented benefits that include inhibition of virulence related gene expression in pathogens and mucosal stimulation of immune cells. The delivery system for probiotics has not been determined for spaceflight, where the food system is shelf stable and the lack of refrigeration prevents the use of traditional dairy delivery methods. In order to demonstrate the potential of the space food system to deliver viable probiotic bacteria to crewmembers, the probiotic L. acidophilus was packaged in high barrier flight packaging in nonfat dry milk (NFDM) or retained in commercial capsule form. Viable cells were enumerated over 8 months of storage at 22, 4, and ‐80ºC. The survival of L. acidophilus rehydrated in NFDM, in a PBS control, and directly from the capsule was also evaluated following stress challenge with simulated gastric and intestinal juices to determine the method that would deliver the most viable cells to the intestine, where they would be expected to confer beneficial effects. L. acidophilus was found to be stable to gastric and intestinal juice challenge when delivered in rehydrated NFDM, even after two hours of exposure. In comparison, L. acidophilus was reduced by 1‐5 logs when exposed to gastric and intestinal juice directly and when rehydrated in a PBS control. Shelf life data indicated that L. acidophilus would require refrigerated or frozen storage to remain viable at adequate levels over the multi‐year storage periods required for spaceflight. This study suggests that the protective effect provided by the dairy matrix, and not merely rehydration prior to consumption, extends probiotic viability and stress tolerance during storage in spaceflight and in simulated digestion conditions more adequately than a capsule. In addition to effective delivery, it is essential to understand the microgravity effects on the stress tolerances and genetic expression of probiotic bacteria to enable optimization of growth, survival, strain selection, and conferred benefits in spaceflight. In our current study, ground‐based characterization of the growth, stress response, and transcriptomic response of L. acidophilus will be accomplished using the low shear modeled microgravity (LSMMG) culture environment in the rotating wall vessel (RWV). We hypothesize that L. acidophilus will maintain or improve its growth and stress response following culture in the LSMMG environment, and that transcriptomic analysis will define the associated molecular mechanism(s), resulting in the ability to optimize strain selection.

Castro, S. L.↗

Sub-melt nanosecond pulsed-laser induced densification and strain-field relaxation in single-crystal diamond

Dislocations and polishing-induced defect networks in synthetic diamond introduce local strain fields and broaden Raman features, limiting performance in optical, thermal, and electronic applications. Laser annealing is emerging as a promising approach to repair surface and near-surface defects in diamond without entering the melt regime, yet surface densification, defect-state modification, and associated structural changes have not been well quantified. In this work, we show that sub-melt nanosecond pulsed-laser annealing (PLA) induces near-surface densification and defect-mediated strain relaxation in single-crystal Chemical Vapor Deposition (CVD) diamond. Single- and two-pulse PLA were applied, and structural evolution was quantified using co-registered ISO 25,178 white-light interferometry, depth-resolved Raman spectroscopy, and cross-sectional STEM with geometric phase analysis (GPA). Across a 5 × 6 grid (n = 30), responsive regions exhibit large reductions in local slope (Sdq 45–65%), developed area (Sdr 60–90%), height spread (Sp, Sz 30–65%), void volume (Vv 57–60%), and roughness amplitude (Sa, Sq 48–57%), consistent with densification of ∼4–6.5 nm. Raman profiling shows narrowing of the diamond line and improved spectral uniformity to depths of ∼2–3 μm. Given that the Raman probing depth significantly exceeds the densified layer thickness, this response is interpreted as consistent with long-range strain-field redistribution originating from the near-surface region. STEM-GPA strain maps further support this interpretation, showing smoother strain fields, suppressed hotspots, and redistribution of localized strain concentrations following PLA. These results are consistent with defect-mediated strain relaxation and densification-driven modification of the near-surface energy state. The approach provides a scalable pathway for improving near-surface structural quality in diamond relevant to electronic, photonic, and quantum applications.

Areal surface metrology (ISO 25,178)↗

The Chlamydomonas Genome Project, version 6: reference assemblies for mating type plus and minus strains reveal extensive structural mutation in the laboratory

Five versions of the Chlamydomonas reinhardtii reference genome have been produced over the last two decades. Here we present version 6, bringing significant advances in assembly quality and structural annotations. PacBio-based chromosome-level assemblies for two laboratory strains, CC-503 and CC-4532, provide resources for the plus and minus mating type alleles. We corrected major misassemblies in previous versions and validated our assemblies via linkage analyses. Contiguity increased over ten-fold and >80% of filled gaps are within genes. We used Iso-Seq and deep RNA-seq datasets to improve structural annotations, and updated gene symbols and textual annotation of functionally characterized genes via extensive manual curation. We discovered that the cell wall-less classical reference strain CC-503 exhibits genomic instability potentially caused by deletion of the helicase RECQ3, with major structural mutations identified that affect >100 genes. We therefore present the CC-4532 assembly as the primary reference, although this strain also carries unique structural mutations and is experiencing rapid proliferation of a Gypsy retrotransposon. We expect all laboratory strains to harbor gene-disrupting mutations, which should be considered when interpreting and comparing experimental results. Collectively, the resources presented here herald a new era of Chlamydomonas genomics and will provide the foundation for continued research in this important reference organism.

59 BASIC BIOLOGICAL SCIENCES↗

Fast growth and high-titer bioproduction from renewable formate via metal-dependent formate dehydrogenase in Escherichia coli

Microbial bioproduction using one-carbon (C1) feedstocks has the potential to decarbonize the manufacturing of materials, fuels, and chemicals. Formate is a promising C1 feedstock, and the realization of industrial, formatotrophic platform organisms is a key goal for C1-based bioproduction. So far, a major limitation for synthetic formatotrophy has been slow energy supply due to slow formate dehydrogenase activity. Here, we implement a fast, metal-dependent formate dehydrogenase complex in a synthetic formatotrophic Escherichia coli utilizing the reductive glycine pathway. After a short-term evolution, we demonstrate formatotrophic growth of E. coli with a doubling time of less than 4.5 h, comparable to the fastest natural formatotrophs. To further explore the potential of a formate-based bioeconomy, this strain is engineered to produce mevalonate, as well as the terpenoid and aviation fuel precursor isoprenol, using formate we generate directly from the electrochemical reduction of CO2. This work demonstrates an improvement in bioproduct titer from formate, achieving the production of 3.8 g/L of mevalonate. Additionally, the abundant and recalcitrant polymer lignin is chemically decomposed into a formate-rich mixture of small organic acids and subsequently bioconverted into mevalonate. Overall, the described fast-growing, formatotrophic bioproduction strain demonstrates that a sustainable formate bioeconomy is within reach.

Cowan, Aidan E↗

Strain-concentration for fast, compact photonic modulation and non-volatile memory

A critical figure of merit (FoM) for electro-optic (EO) modulators is the transmission change per voltage, d T / d V . Conventional approaches in wave-guided modulators maximize d T / d V via a high EO coefficient or longer light-material interaction lengths but are ultimately limited by material losses and nonlinearities. Optical and RF resonances improve d T / d V at the cost of spectral non-uniformity, especially for high- Q optical cavity resonances. Here, we introduce an EO modulator based on piezo-strain-concentration of a photonic crystal cavity to address both trade-offs: (i) it eliminates the trade-off between d T / d V and waveguide loss—i.e., enhancement of the resonance tuning efficiency d v c / d V for the fixed EO coefficient, waveguide length, and cavity Q —and (ii) at high DC strains it exhibits a non-volatile (NV) cavity tuning Δ v c ,NV for passive memory and programming of multiple devices into resonance despite fabrication variations. The device is fabricated on a scalable silicon nitride-on-aluminum nitride platform. We measure d v c / d V =177±1MHz/V, corresponding to Δ v c =40±0.32GHz for a voltage spanning ±120V with an energy consumption of δ U /Δ v c =0.17nW/GHz. The modulation bandwidth is flat up to ω BW,3dB /2 π =3.2±0.07MHz for broadband DC-AC and 142±17MHz for resonant operation near a 2.8 GHz mechanical resonance. Optical extinction up to 25 dB is obtained via Fano-type interference. Strain-induced beam-buckling modes are programmable under a “read-write” protocol with a continuous, repeatable tuning range of 5±0.25GHz, allowing for storage and retrieval, which we quantify with mutual information of 2.4 bits and a maximum non-volatile excursion of 8 GHz. Using a full piezo-optical finite-element-model (FEM) we identify key design principles for optimizing strain-based modulators and chart a path towards achieving performance comparable to lithium niobate-based modulators and the study of high strain physics on-chip.

Wen, Y. Henry (ORCID:0009000685423628)↗

Open Water Blade Strain Measurements on a Vertical-Axis Tidal Turbine

Open-water testing of marine renewable energy devices represents a significant milestone and hurdle for research teams and companies that seek to reduce the levelized cost of energy to allow these devices to compete in the open electrical generation market. For open-water testing of tidal energy converters (TECs), accurate measurements of loading characteristics on the blades and other structural components correlated with power performance metrics can be invaluable to further refine design principles and models, allowing continued improvement and cost reductions of new turbine designs. This paper will describe the modifications and additions made to a TEC system to achieve time-correlated blade strain measurements during full turbine operation along with a discussion on the overall impacts the required modifications had on the unmodified turbine.

mechanical loads↗

Machine-Learning-Based Multiscale Methods for 3D Modelling of Granular Materials by Incorporating History-Dependent State Variables

Over the past decades, the prevalence of machine learning (ML) methods has made the development of ML-based constitutive models for granular materials undoubtedly a popular subject. Numerous studies have been made to feature the loading path or history-dependent stress-strain response of granular media using neural networks. In this work, a novel finite element method (FEM)–ML multiscale approach was developed by incorporating internal variables to improve the simulation accuracy of 3D history-dependent granular materials for the first time. To this end, a surrogate constitutive model based on the single-step-based multi-layer perceptron (MLP) neural network was used to replace representative volume element (RVE) simulations conducted by the discrete element method (DEM) in the multiscale FEM–DEM approach. Although the prediction principle of the MLP aligns with the FEM algorithm, artificially added internal variables are required to differentiate the loading history. To address this issue, history variables associated with the Frobenius norm are proposed to be fed into the MLP coupled with the strain tensor to extract the history-dependent behaviour of granular assemblies. The developed FEM–ML approach was demonstrated in 3D conventional triaxial compression (CTC) simulations. Compared to the multiscale FEM–DEM approach, the proposed FEM–ML method exhibits a significantly improved computational efficiency.

granular materials↗

Sharply notch cylindrical tension specimen for screening plane-strain fracture toughness. I - Influence of fundamental testing variables on notch strength. II Applications in aluminum alloy quality assurance of fracture toughness

A description is presented of studies which have been conducted to establish an improved technology base for a use of the sharply notched cylindrical specimen in quality assurance tests of aluminum alloy products. The results are presented of an investigation of fundamental variables associated with specimen preparation and testing, taking into account the influence of the notch root radius, the eccentricity of loading, the specimen diameter, and the notch depth on the sharp notch strength. Attention is given to the statistical procedures which are necessary to establish correlations between the sharp notch strength and the plane-strain fracture toughness for high-strength aluminum alloys.

Jones, M. H.↗

Developing a robust strength model using physically-informed genetic programming

The strength of materials is influenced by a range of external conditions, such as temperature and deformation rate. Consequently, materials that demonstrate substantial variations in their mechanical behavior due to fluctuations in temperature and strain rate require complex strength models to accurately predict material performance in real-world applications. To predict such complex behavior, a robust and flexible strength model is necessary. In this work, we utilize genetic programming-based symbolic regression (GPSR) to develop data-driven strength models that accurately represent the measured stress–strain responses of tin across a wide range of strain, strain rate and temperature regimes. The GPSR models are constrained by physically-informed conditions, which leads to significant improvement in extrapolation. The best model is integrated into a multi-physics code to perform Taylor impact simulations, validating the model’s accuracy and robustness. In conclusion, the model predictions showed excellent agreement with experimental results, particularly when compared to predictions using traditional strength models.

Genetic programming↗

3D-printing methacrylate/chitin nanowhiskers composites via stereolithography: Mechanical and thermal properties

We report on the mechanical and thermal properties of stereolithography (SLA) 3D-printed methacrylate (MA) resin reinforced by chitin nanowhiskers (CNWs). CNWs were synthesized by acid hydrolysis of crab shell chitin and nanocomposites with 0 wt%, 0.5 wt%, 1.0 wt%, and 1.5 wt% CNWs loading were prepared by slurry compounding method. The obtained CNWs (diameter = 23 ± 5 nm, length = 253 ± 100 nm, and aspect ratio = 11 ± 3) are well dispersed within the bulk of MA at 1.0 wt% loading, implying good interfacial adhesion between the nanofiller and the resin matrix possibly due to hydrogen bonding interactions. The addition of up to 1.0 wt% CNWs to neat MA enhances the tensile strength, strain at break, modulus, and maximum thermal degradation temperatures of the nanocomposites. Moreover, the incorporation of CNWs not only improves the mechanical and thermal properties, but also preserves the excellent resolution and accuracy of 3D-printed MA/CNWs nanocomposites.

36 MATERIALS SCIENCE↗

Demonstration and technoeconomic analysis of dodecanol production from acetate using metabolically engineered Escherichia coli

In a circular bioeconomy, the one-way conversion of petroleum to chemicals and CO 2 is replaced with processes that reduce CO 2 to energy carriers and useful materials that are returned to CO 2 upon combustion. A circular bioeconomy that relies on photosynthesis to generate sugars as the chief energy carrier and precursor to chemical building blocks has yet to overcome many recalcitrant aspects of plant-based photosynthesis, namely, high feedstock costs, arable land scarcity, food competition, and fertilizer overuse. Acetate is a potential sustainable energy carrier because it can be produced from CO 2 either electrocatalytically or by acetogens via the Wood-Ljungdahl pathway. Here, in this work, we conducted a metabolic engineering study of Escherichia coli 's ability to convert acetate into dodecanol as a model oleochemical product. We performed techno-economic and life cycle analyses to determine break-even points with alternative fossil fuel-based strategies and identified critical process performance parameters for supporting an industrial acetate-based bioprocess. These analyses showed that oleochemical yield is the primary driver of minimum oleochemical selling price and carbon intensity. Therefore, to increase yield on acetate, we deleted the aceBAK operon, which facilitates funneling of acetate into biomass instead of product. We performed additional strain engineering to increase flux towards dodecanol and increase acetate uptake. Finally, we demonstrated increased yield in controlled bioreactors, improving from 13% of the maximum theoretical yield to 37%. Rigorous uncertainty analyses assuming a range of market conditions and future technological performances resulted in 88% and 37% of simulated scenarios having lower carbon intensities than fossil fuel-based routes and lower minimum selling prices than the market price.

Acetate↗

Monolayer Sc 2 CF 2 as a Potential Selective and Sensitive NO 2 Sensor: Insight from First-Principles Calculations

Two-dimensional materials with excellent surface–volume ratios and massive reaction sites recently have been receiving attention for gas sensing. With first-principles calculations, we explored the performance of monolayer Sc 2 CF 2 as a gas sensor. We investigated how molecule adsorption affects its electronic structure and optical properties. It is found that a large charge transfer quantity happens between Sc 2 CF 2 and NO 2 , which results from the fact that the lowest unoccupied molecular orbital (LUMO) of NO 2 is below the valence band maximum (VBM) of Sc 2 CF 2 . Moreover, the MD simulation shows that NO 2 can adsorb on the Sc 2 CF 2 surface stably at room temperature. We explored the effect of biaxial strain on the adsorption energy and charge transfer quantity of each system, and the results show that the biaxial strain can enhance both the adsorption energy and charge transfer quantity of the NO 2 system and thus can improve the sensitivity of Sc 2 CF 2 in detecting the NO 2 molecule. Furthermore, we investigated the adsorption behavior and charge transfer of polar polyatomic molecules at the Sc 2 CF 2 surface with h-BN as a substrate, and the results demonstrate that the h-BN substrate can hardly modify the main results. Our result predicts that Sc 2 CF 2 can be a promising selective and sensitive sensor to detect the NO 2 molecule, and could also give a theoretical guide for other terminated MXenes used for gas sensors or detectors.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Cryogenic aspects of a 20 MW class low-temperature superconducting generator for the renewables industry

In this paper we give a progress update on the cryogenic design for cooling a 20 MW class, partially superconducting generator with stationary field coils for offshore wind renewables industry [1-2]. This is a continuation of an earlier program on a 10 MW system dating back ten years. Whereas this new power rating increase leads to a radial diameter expansion of the superconducting field coils from 4 to > 9.5 m, it maintains the axial length. We show the design based on this scaled up with enlarged diameter. The field coil size increase asks for higher cooling power that leads to a bigger cold box size to accommodate the cryocoolers. In the design, as many as 8 cryocoolers can be accessed and serviced from the nacelle that houses the main cryogenic components. A typical thermal load balance sheet with all components is given and compared against the available cryocooler cooling power at different operating conditions. Due to the cryocoolers’ local point of contact cooling within the nacelle, the temperature gradient of the thermal shield that fully encloses the cold mass with its embedded field coils needs to be balanced out to minimize the heat burden on the field coils. This requires additional analytical efforts and implementation of further design features. The extended nacelle houses the cold box with its cryogenic infrastructure. The interface for the envisaged cryogenic pushbutton closed-loop circulating system remains invisible, requires no handling of cryogenic liquids and is hermetically closed. The field coil diameter increase also leads to a greater initial helium gas storage volume. In addition, it requires higher initial room temperature fill pressure within the toroidal helium vapor storage tanks and in cooling tubes connecting to the cryocoolers. The toroidal helium gas tanks are thermally coupled with the thermal shield so that helium convection inside the storage tank can improve heat transfer and reduce the temperature gradient of the thermal shield. Besides those heat transfer challenges, additional mechanical strain within this large structure is exerted on the torque tubes during initial cooldown and when energizing field coils. Some of those design challenges are quite unexpected, leading to novel workarounds in order to maintain the chosen cooling strategy. Finally, we assess those design limitations in view of further cryogenic scalability with emphasis on manufacturability and assembly.

cryogenics↗

Thin-film lithium niobate on diamond as a platform for efficient spin-phonon coupling

Negatively charged silicon vacancy (SiV − ) centers in diamond are leading candidates for solid-state quantum memory that can be controlled using electromagnetic or acoustic waves. The latter are particularly promising due to the strong strain response of SiV − , enabling large spin-phonon interaction strengths. Indeed, coherent spin control via surface acoustic waves (SAWs) has been demonstrated and is essential for developing on-chip phononic quantum networks. However, the absence of piezoelectricity in diamond requires an interface with a piezoelectric material for efficient transduction and delivery of acoustic waves to spins in diamonds. Here, we demonstrate a heterogeneously integrated phononic platform that combines thin-film lithium niobate with diamond to enable acoustic control of single SiV − spins. Additionally, leveraging large SAW-induced strain at the location of SiV − , we achieve coherent acoustic control of an electron spin with more than twofold improvement in Rabi frequency compared to state-of-the-art devices based on aluminum nitride on diamond. Furthermore, this work represents a crucial step towards realizing phonon-based quantum information processing systems.

Optically detected magnetic resonance↗

Structural analysis of the NifL-NifA complex reveals the molecular basis of anti-activation of nitrogen fixation gene expression in Azotobacter vinelandii

Understanding the molecular basis of regulated nitrogen (N 2 ) fixation is essential for engineering N 2 -fixing bacteria that fulfill the demand of crop plants for fixed nitrogen, reducing our reliance on synthetic nitrogen fertilizers. In Azotobacter vinelandii and many other members of Proteobacteria, the two-component system comprising the anti-activator protein (NifL) and the Nif-specific transcriptional activator (NifA)controls the expression of nif genes, encoding the nitrogen fixation machinery. The NifL-NifA system evolved the ability to integrate several environmental cues, such as oxygen, nitrogen, and carbon availability. The nitrogen fixation machinery is thereby only activated under strictly favorable conditions, enabling diazotrophs to thrive in competitive environments. While genetic and biochemical studies have enlightened our understanding of how NifL represses NifA, the molecular basis of NifA sequestration by NifL depends on structural information on their interaction. Here, we present mechanistic insights into how nitrogen fixation is regulated by combining biochemical and genetic approaches with a low-resolution cryo-electron microscopy (cryo-EM) map of the oxidized NifL-NifA complex. Our findings define the interaction surface between NifL and NifA and reveal how this interaction can be manipulated to generate bacterial strains with increased nitrogen fixation rates able to secrete surplus nitrogen outside the cell, a crucial step in engineering improved nitrogen delivery to crop plants.

59 BASIC BIOLOGICAL SCIENCES↗