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Cogeneration Technology Alternatives Study (CTAS). Volume 6: Computer data. Part 1: Coal-fired nocogeneration process boiler, section B

About fifty industrial processes from the largest energy consuming sectors were used as a basis for matching a similar number of energy conversion systems that are considered as candidate which can be made available by the 1985 to 2000 time period. The sectors considered included food, textiles, lumber, paper, chemicals, petroleum, glass, and primary metals. The energy conversion systems included steam and gas turbines, diesels, thermionics, stirling, closed cycle and steam injected gas turbines, and fuel cells. Fuels considered were coal, both coal and petroleum based residual and distillate liquid fuels, and low Btu gas obtained through the on site gasification of coal. Computer generated reports of the fuel consumption and savings, capital costs, economics and emissions of the cogeneration energy conversion systems (ECS's) heat and power matched to the individual industrial processes are presented. National fuel and emissions savings are also reported for each ECS assuming it alone is implemented. Two nocogeneration base cases are included: coal fired and residual fired process boilers.

Knightly, W. F.↗

Photo-Induced Deuterium Enrichment in Residues Produced from the UV Irradiation of Pyrimidine in H2O and H2O+NH3 Ices

Organic compounds found in meteorites often show isotopic signatures of their interstellar/protosolar heritage as enrichments in D and 15N. Meteoritic organics found to be enriched in D include amino acids, hydroxy and dicarboxylic acids, as well as polycyclic aromatic hydrocarbons (PAHs). Processes that can produce isotopic enrichments in presolar/protosolar materials include gas-phase ion-molecule reactions, gas-grain surface reactions, and unimolecular photo-dissociation reactions involving PAHs. Because many molecules in interstellar clouds are enriched in D, the presence of D anomalies in meteorites is thought to originate from preserved or slightly altered interstellar/protostellar materials. However, the link between isotopic enrichments seen in space and those in meteoritic compounds and their relationship remain unclear. In this work, we present results of hydrogen isotopic fractionation for compounds in organic residues produced from the UV irradiation using an H2- discharge UV lamp of H2O:pyrimidine = 20:1 and H2O:NH3:pyrimidine = 20:2:1 ice mixtures at low temperature (is less than 20 K). After irradiation, the resulting residues are dissolved in H2O and analyzed with gas chromatography-mass spectrometry coupled with isotope ratio mass spectrometry (GC-MS/IRMS) [1], following a protocol similar to that used for previous analyses of comparable samples [2,3]. We used this technique to measure compound-specific D/H isotopic ratios for the initial pyrimidine and for two photo-products present in the residues, namely, 2,2'-bipyrimidine and an unidentified bipyrimidine isomer [2-4]. Measuring D enrichments in bipyrimidines has the advantage that the H atoms on these molecules are not easily exchangeable with other compounds, in particular the H2O and NH3 present in the ices or the solvents used to extract the samples for GC-MS/IRMS measurements. The δD value for the initial pyrimidine, measured with a high-temperature conversion elemental analyzer connected to the IRMS, was found to be -30% per mille. Preliminary measurements made on a residue produced from the UV irradiation of an H2O:NH3:pyrimidine = 20:2:1 ice mixture indicate δD values of +118% per mille for 2,2'- bipyrimidine and +92% per mille for the other bipyrimidine isomer, and therefore show a significant D enrichment during the photo-processing and warm-up that lead to their formation [5]. New measurements are currently being performed on a number of residues produced from simpler H2O:pyrimidine = 20:1 ice mixtures under different experimental conditions and will be presented here.

Laboratory Astrochemistry↗

Biopower: Impact of Biofuels Deployment to Replace Petroleum Liquids in Stationary Power Applications

Petroleum-based liquids are used in a portion of power generation applications in the United States, predominantly in the New England, Middle Atlantic, South Atlantic, and Pacific-Noncontiguous regions. Power plants that burn petroleum liquids, such as distillate or residual fuel oils, are generally used for short periods to accommodate peak electricity demands. The Energy Information Administration (EIA) estimated the U.S. consumption of petroleum liquids for electricity generation at 27 million barrels in 2018, representing a cost of $2.4 billion annually. This study assesses the potential to displace all or part of the petroleum liquids in U.S. power generation with biofuels. The biofuels for this application are assumed to be derived from terrestrial feedstocks, with conversion routes of both fast pyrolysis (bio-oil) and hydrothermal liquefaction (bio-crude). Regional models were used to assess the availability and cost of three different base materials: clean wood, forest residues, and corn stover; each was evaluated in the laboratory at small or experimental scales for conversion to bio-oil or bio-crude. The estimated biofuel production quantities depend on equivalent heating versus the current heavy fuel. In this report, the availability of each type of biomass for each section of the U.S. Census division is estimated using a conservative broker price (in each case) of $ 80 per dry tonne. The results show that the petroleum-liquid power generation in each of the Census Divisions could be supplied by one or more of the feedstocks evaluated. For all regions, clean wood supplies (only) could provide ample supply. For all but two regions (Middle Atlantic and New England), forest residues alone are sufficient. Finally, for all regions but three (Middle Atlantic, New England, and South Atlantic), corn stover alone is adequate. The Minimum Fuel Selling Price (MFSP) of bio-oil and bio-crude were also estimated for each feedstock type and Census Division. This analysis showed that fast pyrolysis bio-oil projections to be lower (14% on average) than current wholesale petroleum-based heating oil prices in each of the regions, assuming 100 dry tonnes/day processing capacity. However, bio-crude predictions were significantly higher (2X) in all cases. The effect of biorefinery size was also quantified. Based on the preliminary results in this study, it is apparent the biofuels could be an economical alternative for current petroleum liquids in U.S. power generation. However, additional research is needed to determine the necessary biofuel characteristics to support existing generation equipment. It is recommended that both power generation and biofuel production stakeholders to be engaged to outline the research and testing needed to identify the technical hurdles to enable the opportunity.

02 PETROLEUM↗

Genetically engineered poplar wood effectively enhances the efficiency of deep eutectic solvent-mediated one-pot processing

Although lignocellulosic biomass is a renewable resource with the potential to replace fossil-derived fuels and chemicals, its recalcitrance, largely due to lignin, limits its utilization. Recent advancements in genetic engineering have produced transgenic trees with reduced lignin content and/or modified lignin structure without compromising growth traits. Here, three engineered poplar varieties are evaluated as feedstocks using a biocompatible one-pot deep eutectic solvent-mediated process that integrates biomass fractionation and enzymatic saccharification within a single reactor, eliminating water washing and reconditioning. All transgenic poplars exhibit higher fermentable sugar yields than wild-type (WT) trees. Notably, QsuB poplar, incorporating 3,4-dihydroxybenzoate in lignin, achieves the highest glucose conversion yield of 91.3% (vs. 73.0% from WT). AT5 and MdCHS3 poplars, incorporating ferulate esters and naringenin, also demonstrate improved glucose yields (86.7 and 84.7%, respectively), confirming reduced biomass recalcitrance. Additionally, residual lignins are valorized via hydrogenolysis into phenolic compounds, with comparable alkylphenol production across all lines. These findings demonstrate that the transgenic poplar lines not only serve as superior feedstocks for sugar conversion but also provide a rich resource for phenolic compound production, enhancing the operational and economic viability of integrated biorefinery processes.

biomasses↗

Generative Framework Approach to Match Landsat and Sentinel-2 Data

Linear regression and histogram matching based techniques have been widely used to minimize the surface reflectance difference between two similar satellite observations such as Landsat-8/9 and Sentinel-2A/B products [1]. However, regionally or globally derived conversion factors may not be suitable for all land cover types and locations, resulting in noticeable residual differences between the sensors. Generative Adversarial Network (GAN) has shown promise in the field of image processing for domain or style transfer[2]. In this work we aim to minimize the surface reflectance difference between Landsat and Sentinel-2 products based on GAN.

Sujit Roy↗

Molecular information theory meets protein folding

We propose an application of molecular information theory to analyze the folding of single domain proteins. We analyze results from various areas of protein science, such as sequence-based potentials, reduced amino acid alphabets, backbone configurational entropy, secondary structure content, residue burial layers, and mutational studies of protein stability changes. We found that the average information contained in the sequences of evolved proteins is very close to the average information needed to specify a fold ~2.2 ± 0.3 bits/(site operation). The effective alphabet size in evolved proteins equals the effective number of conformations of a residue in the compact unfolded state at around 5. We calculated an energy-to-information conversion efficiency upon folding of around 50%, lower than the theoretical limit of 70%, but much higher than human built macroscopic machines. We propose a simple mapping between molecular information theory and energy landscape theory and explore the connections between sequence evolution, configurational entropy and the energetics of protein folding.

Ignacio E. Sánchez↗

Cogeneration Technology Alternatives Study (CTAS). Volume 6: Computer data. Part 1: Coal-fired nocogeneration process boiler, section A

Various advanced energy conversion systems (ECS) are compared with each other and with current technology systems for their savings in fuel energy, costs, and emissions in individual plants and on a national level. About fifty industrial processes from the largest energy consuming sectors were used as a basis for matching a similar number of energy conversion systems that are considered as candidates which can be made available by the 1985 to 2000 time period. The sectors considered included food, textiles, lumber, paper, chemicals, petroleum, glass, and primary metals. The energy conversion systems included steam and gas turbines, diesels, thermionics, stirling, closed cycle and steam injected gas turbines, and fuel cells. Fuels considered were coal, both coal and petroleum based residual and distillate liquid fuels, and low Btu gas obtained through the on-site gasification of coal. Computer generated reports of the fuel consumption and savings, capital costs, economics and emissions of the cogeneration energy conversion systems (ECS's) heat and power matched to the individual industrial processes are presented for coal fired process boilers. National fuel and emissions savings are also reported for each ECS assuming it alone is implemented.

Knightly, W. F.↗

A broad specificity β-propeller enzyme from Rhodopseudomonas palustris that hydrolyzes many lactones including γ-valerolactone

Lactones are prevalent in biological and industrial settings, yet there is a lack of information regarding enzymes used to metabolize these compounds. One compound, γ-valerolactone (GVL), is used as a solvent to dissolve plant cell walls into sugars and aromatic molecules for subsequent microbial conversion to fuels and chemicals. Despite the promise of GVL as a renewable solvent for biomass deconstruction, residual GVL can be toxic to microbial fermentation. Here, we identified a Ca 2+ -dependent enzyme from Rhodopseudomonas palustris (Rpa3624) and showed that it can hydrolyze aliphatic and aromatic lactones and esters, including GVL. Maximum-likelihood phylogenetic analysis of other related lactonases with experimentally determined substrate preferences shows that Rpa3624 separates by sequence motifs into a subclade with preference for hydrophobic substrates. Additionally, we solved crystal structures of this β-propeller enzyme separately with either phosphate, an inhibitor, or a mixture of GVL and products to define an active site where calcium-bound water and calcium-bound aspartic and glutamic acid residues make close contact with substrate and product. Our kinetic characterization of WT and mutant enzymes combined with structural insights inform a reaction mechanism that centers around activation of a calcium-bound water molecule promoted by general base catalysis and close contacts with substrate and a potential intermediate. Similarity of Rpa3624 with other β-propeller lactonases suggests this mechanism may be relevant for other members of this emerging class of versatile catalysts.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Low NO/x/ and fuel flexible gas turbine combustors

The feasibility of various low NO(x) emission gas turbine combustor configurations was evaluated. The configurations selected for fabrication and testing at full pressure and temperature involved rich-lean staged combustion utilizing diffusion flames, rich-lean prevaporized/premix flames, and staged catalytic combustion. The test rig consisted of a rich burner module, a quench module, and a lean combustion module. Test results are obtained for the combustor while burning petroleum distillate fuel, a coal derived liquid, and a petroleum residual fuel. The results indicate that rich-lean diffusion flames with low fuel-bound nitrogen conversion are achievable with very high combustion efficiencies.

Lew, H. G.↗

Evaluation of Enzymatic Hydrolysis of CELSS Wheat Residue Cellulose at a Scale Environment to NASA's KSC Breadboard Project

Biomass processing at the Kennedy Space Center CELSS breadboard project has focused on the evaluation of breadboard-scale enzymatic hydrolysis of wheat residue cellulose (25%, w/w). Five replicate runs of cellulase production by Trichoderma reesei (QM9414) and enzymatic hydrolysis of residue cellulose were completed. Enzymes were produced in 1 0 days (5 L, 25 g (dry weight) residue). Cellulose hydrolysis (12 L, 50 g (dry weight) residue) using these enzymes produced 5.5 to 6.0 g glucose liter(exp -1) in 7 days. Cellulose conversion efficiency was 29%. These processes are feasible technically on a breadboard scale, but would only increase the edible wheat yield 10%.

Strayer, Richard F.↗

Global and Seasonal Aerosol Optical Depths Derived From Ultraviolet Observations by Satellites (TOMS)

It has been shown that absorbing aerosols (dust, smoke, volcanic ash) can be detected in the ultraviolet wavelengths (331 nm to 380 nm) from satellite observations (TOMS, Total Ozone Mapping Spectrometer) over both land and water. The theoretical basis for these observations and their conversions to optical depths is discussed in terms of an aerosol index AI or N-value residue (assigned positive for absorbing aerosols). The theoretical considerations show that negative values of the AI frequently represent the presence of non-absorbing aerosols (NA) in the troposphere (mostly pollution in the form of sulfates, hydrocarbons, etc., and some natural sulfate aerosols) with particle sizes near 0.1 to 0.2 microns or less. The detection of small-particle non-absorbing aerosols from the measured backscattered radiances is based on the observed wavelength dependence from Mie scattering after the background Rayleigh scattering is subtracted. The Mie scattering from larger particles, 1 micron or more (e.g., cloud water droplets) has too small a wavelength dependence to be detected by this method. In regions that are mostly cloud free, aerosols of all sizes can be seen in the single channel 380 nm or 360 nm radiance data. The most prominent Al feature observed is the strong asymmetry in aerosol amount between the Northern and Southern Hemispheres, with the large majority of NA occurring above 20degN latitude. The maximum values of non-absorbing aerosols are observed over the eastern U.S. and most of western Europe corresponding to the areas of highest industrial pollution. Annual cycles in the amount of NA are observed over Europe and North America with maxima occurring in the summer corresponding to times of minimum wind transport. Similarly, the maxima in the winter over the Atlantic Ocean occurs because of wind borne transport from the land. Most regions of the world have the maximum amount of non-absorbing aerosol in the December to January period except for the eastern North America and Europe. Comparisons of the estimated TOMS aerosol optical depths show good agreement in magnitude and seasonal dependence with sun-photometer optical depths obtained at Goddard Space Flight Center (39degN 76.88degW) in the U.S. and in Lille (50.63degN 3.07degE) in France. The study of these aerosols is important for detecting the sources of industrial pollution and its redistribution by winds on a global basis, as well as its effect on reducing the UV irradiance at the Earth's surface.

Herman, J. R.↗

BETO 2021 Peer Review - Rational Design of Robust Reactor Feeding Systems for Heterogeneous Cellulosic and Agricultural Wastes Based on Biomass Quality Characteristics

Consistent and reliable preprocessing, conveyance, and reactor in-feed systems, particularly for low-cost waste feedstocks, remains a major technical challenge for the emerging Bioeconomy. By identifying critical biomass attributes and connecting them to flow and conversion behavior, science-driven system designs can address these often-overlooked solids handling challenges. The Wonderful Company (TWC) is the world's largest almond and pistachio grower, generating 250,000 dry tons/year of waste material including hulls, shells, and wood (>5 million tons/year industry wide in the U.S.). This project seeks to turn this environmental and economic liability into a sustainable and profitable resource, targeting conversion via gasification to syngas for electricity and bio-char, by addressing related material handling and feeding challenges. Based on optimized preprocessing strategies, bulk material flow, and thermal conversion properties, an overall system design will be developed. The methodology will be tested with FCIC's benchmark loblolly pine residues, to demonstrate the robustness of the overall approach and provide insight and guidance for future systems. The project will culminate in extended field trials to demonstrate an improved continuous feeding system with a commercial biomass-to-electricity gasifier vendor, and an economic analysis demonstrating a reduction in electricity production costs by maximizing on-stream time while minimizing preprocessing and CapEx costs.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Biomass Attributes and Attribute Modifications Affecting Systems and Methods to Separate and Fractionate

Chemical and physical heterogeneity in biomass feedstocks such as agricultural or forestry residues is due to substantial differences in plant tissue types. These differences can contribute significant challenges to handling, preprocessing, and conversion in biorefining processes. An understanding of this chemical and physical heterogeneity can be used to inform fractionation technologies that could facilitate more streamlined processing and potentially be employed to yield multiple co-product streams for a single feedstock. In this chapter, the motivation and scope of biomass fractionation is first outlined. Physical and chemical properties of biomass feedstocks, along with their distribution and diversity within plants, are next discussed with respect to how these differences can be exploited in a fractionation process. A summary of some of the key physical principles that allow for fractionation is next covered along with how these physical principles are exploited in equipment designs. Examples from the literature are briefly discussed that highlight how these approaches can be employed to achieve processing objectives. Several case studies on physical fractionation of corn stover and forestry residues are presented that illustrate how integrated fractionation processes could be employed. Lastly, prospects and potential economic drivers for adoption of biomass fractionation technologies are discussed.

09 BIOMASS FUELS↗

Biomass Attributes and Attribute Modifications Affecting Systems and Methods to Separate and Fractionate. In: Handbook of Biorefinery Research and Technology

Chemical and physical heterogeneity in biomass feedstocks such as agricultural or forestry residues is due to substantial differences in plant tissue types. These differences can contribute significant challenges to handling, preprocessing, and conversion in biorefining processes. An understanding of this chemical and physical heterogeneity can be used to inform fractionation technologies that could facilitate more streamlined processing and potentially be employed to yield multiple co-product streams for a single feedstock. In this chapter, the motivation and scope of biomass fractionation is first outlined. Physical and chemical properties of biomass feedstocks, along with their distribution and diversity within plants is next discussed with respect to how these differences can be exploited in a fractionation process. A summary of some of the key physical principles that allow for fractionation is next covered along with how these physical principles are exploited in equipment designs. Examples from the literature are briefly discussed that highlight how these approaches can be employed to achieve processing objectives. Several case studies on physical fractionation of corn stover and forestry residues are presented that illustrate how integrated fractionation processes could be employed. Finally, prospects and potential economic drivers for adoption of biomass fractionation technologies are discussed.

Biomass chemical properties↗

Real Time Observation of Lithium Insertion into Pre-Cycled Conversion-Type Materials

Conversion-type electrode materials for lithium-ion batteries experience significant structural changes during the first discharge–charge cycle, where a single particle is taken apart into a number of nanoparticles. This structural evolution may affect the following lithium insertion reactions; however, how lithiation occurs in pre-cycled electrode materials is elusive. In this work, in situ transmission electron microscopy was employed to see the lithium-induced structural and chemical evolutions in pre-cycled nickel oxide as a model system. The introduction of lithium ions induced the evolution of metallic nickel, with volume expansion as a result of a conversion reaction. After pre-cycling, the phase evolutions occurred in two separate areas almost at the same time. This is different from the first lithiation, where the phase change takes place successively, with a boundary dividing the reacted and unreacted areas. Structural changes were restricted at the areas having large amount of fluorine, implying the residuals from the decomposition of electrolytes may have hindered the electrochemical reactions. This work provides insights into phase and chemical evolutions in pre-cycled conversion-type materials, which govern electrochemical properties during operation.

25 ENERGY STORAGE↗

Synthesis‐Related Nanoscale Defects in Mo‐Based Janus Monolayers Revealed by Cross‐Correlated AFM and TERS Imaging

2D Janus transition metal dichalcogenides (TMDs) are promising candidates for various applications including non-linear optics, energy harvesting, and catalysis. These materials are usually synthesized via chemical conversion of pristine TMDs. Nanometer-scale characterization of the obtained Janus materials’ morphology and local composition is crucial for both the synthesis optimization and the future device applications. In this work, we present the results of cross-correlated atomic force microscopy (AFM) and tip-enhanced Raman spectroscopy (TERS) study of Janus monolayers synthesized by the hydrogen plasma-assisted chemical conversion of MoSe 2 and MoS 2 . We demonstrate that the choice of both the growth substrate and the starting TMD influences the residual strain, thereby shaping the nanoscale morphology of the resulting Janus material. Furthermore, by employing TERS imaging, we show the presence of nanoscale islands (≈20 nm across) of MoSe 2 - ${\mathrm{Mo}}_{{\mathrm{Se}}}^{\mathrm{S}}$ (MoS 2 -${\mathrm{Mo}}_{\mathrm{S}}^{{\mathrm{Se}}}$) vertical heterostructures originating from the bilayer nanoislands in the precursor monolayer crystals. The understanding of the origins of nanoscale defects in Janus TMDs revealed in this study can help with further optimization of the Janus conversion process towards uniform and wrinkle-/crack-free Janus materials. Moreover, this work shows that cross-correlated AFM and TERS imaging is a powerful and accessible method for studying nanoscale composition and defects in Janus TMD monolayers.

2D materials↗

Woody Feedstock 2022 State of Technology Report

The U.S. Department of Energy promotes production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the state of technology (SOT). As part of its involvement in this mission, Idaho National Laboratory completes an annual SOT report for n th -plant and 1 st -plant woody biomass feedstock logistics. The purpose of the SOT is to provide the status of feedstock supply system technology development for woody biomass to biofuels relative to technical targets and cost goals from specific design cases, based on data and experimental results. Conventional feedstock supply systems need to be modified to meet the demands of conversion pathways, specifically to have the ability to adjust the quality of the raw biomass materials. Advanced systems incorporate innovative methods of material handling, preprocessing and supply chain configuration. In advanced designs, variability of the raw biomass can be reduced to produce feedstocks of a uniform format, moving toward biomass commoditization. Against this backdrop, the 2022 Woody SOT for low-ash woody feedstocks utilizes feedstock fractionation by incorporating technologies that can separate the biomass into its anatomical fractions (wood, bark, needle, and extrinsic ash) to reduce impurities and attempt to maximize the retention of usable fractions that satisfy downstream quality considerations. By using a series of air classification steps, this strategy can reduce the extrinsic ash in forest residues, separate out a majority of the incoming needles (which can be supplied to alternate markets), and maximize the retention of whitewood in the usable fraction. The fractionated forest residues are then mixed with clean-pine chips in a 50-50 blend to prepare the feedstock for the desired conversion pathway. The n th -plant analysis estimated the delivered cost for the feedstock at $\$$69.23/dry ton (2016$\$$) which represents a $\$$6.64/dry ton decrease compared to the cost estimate of the 2021 Woody SOT supply system for low-ash woody feedstocks. The quality requirements in the 2022 Woody SOT were identical to those of the 2021 Woody SOT at = 1.00 wt % ash and = 50.51 wt% carbon. The cost savings derive primarily from reductions in dry matter losses during air classification. The GHG emissions for the n th -plant analysis were estimated at 178.39 kg CO2e/dry ton compared to 178.71 kg CO 2 e/dry ton in the 2021 Woody SOT, a decrease of 0.32 kg CO2e/dry ton. The small change stems from an increase in emissions attributed to preprocessing and slightly larger savings in emissions from transportation. In the 1 st -plant analysis of the 2022 Woody SOT system, the average throughput was estimated to be approximately 2,128 dry tons/day or 96.51% of the name plate capacity. During the simulation the daily throughput ranged from 1,090 dry tons/day to 2,200 dry tons/day, or 49.43% to 99.75% of the daily nameplate capacity. After the year of operation 722,403 tons of processed feedstock were produced in total without regard to quality considerations (99.64% of the annual nameplate capacity). The variability in throughput was primarily caused by equipment failures in the system. Regular failures, downtime caused by routine maintenance per manufacturer guidelines, contributed to a majority 62.50% of failures and 62.60% of downtime. Failures due to wear were the other cause of disruption within the system, impacting the rotary shear and orbital screen and accounting for 37.50% of the failures and 37.40% of the total downtime. Ultimately the system was on stream for 87.84% during the simulation period, which is only 2.16 percentage points below the nth-plant assumption for on-stream time. The production cost of the system averaged $\$$71.66/dry ton. The costs ranged from a minimum of $\$$71.23/dry ton to a maximum of $\$$2,115.30/dry ton. When dry matter losses (disposed low-quality fractions as well as other losses such as in grinders) were considered the costs increased to an average of $\$$75.11/dry ton with a minimum of $\$$74.69/dry ton and a maximum of $\$$2,136.86/dry ton...

09 BIOMASS FUELS↗

Trafficking of a nitrogenase FeMo-cofactor assembly intermediate

The maturation of the unique FeMo-cofactor of molybdenum nitrogenase is a multistep process requiring the sequential action of a series of maturase complexes. As a final step, the NifEN complex forms FeMo-cofactor from the precursor NifB-co, also called L-cluster, through replacement of an apical iron ion by molybdenum and the attachment of an organic homocitrate ligand. NifB-co is delivered by a small cofactor chaperone, NifX, and initially bound near the surface of the maturase NifEN. Here, we report high-resolution cryo-electron microscopy structures of NifEN in complex with NifX, showing NifB-co binding to NifEN in full detail, capturing both interacting partners in the act of cluster transfer. In a dynamic transfer complex, the large metal cluster is coordinated by single residues from both NifEN and NifX. In silico studies concur with these structures but suggest a third, internal conversion site where cluster maturation likely takes place.

metalloproteins↗