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At least 343 records · Page 19

Carbothermal Reduction Demonstration (CaRD) Gas Analysis Subsystem Development

The Carbothermal Reduction Demonstration (CaRD) project is currently developing a subscale system to demonstrate the operation and performance of the carbothermal reduction process and quantify the production of molar equivalent oxygen (carbon monoxide (CO) and carbon dioxide (CO2)). Traditionally, gas chromatography (GC) systems are utilized to qualify gases due to their ability to separate the volatile compounds within the gas mixture, followed by quantification of the isolated component of interest. Despite these capabilities, GCs require consumables for their operational mobile phase and consequently are not often considered a flight-forward option. An alternative analytical technique known as mass spectrometry measures the presentence of gas phase molecules based on their mass-to-charge ratio. Residual gas analyzers (RGAs) which utilize mass spectrometry are not traditionally used to quantify analytes, but proper design and calibrations can result in a flight-forward analytical instrument capable of direct quantification of volatile gases of interest. Kennedy Space Center's role for the project is to design a gas analyzer system to detect and quantify the CO and CO2 gas produced during the carbothermal reaction utilizing a commercial version of the Mass Spectrometer Observing Lunar Operations (MSolo) instrument. MSolo is a modified commercial off-the-shelf mass spectrometer which consists of quadrupole mass filter and space rated electronics. MSolo is currently a TRL 6 technology and manifested on the PRIME-1 and VIPER missions. This paper describes leveraging MSolo based instrumentation to develop a gas analysis system for the work outlined in the 2023 International Conference on Environmental Systems (ICES) paper number 313. The design and testing of the brassboard phase of the project will be described along with the current work underway which will cover the design and preliminary test results of the prototype phase.

Ryan Patrick Gott↗

Carbothermal Reduction Demonstration (CaRD) Gas Analysis Subsystem Development

The Carbothermal Reduction Demonstration (CaRD) project is currently developing a subscale system to demonstrate the operation and performance of the carbothermal reduction process and quantify the production of molar equivalent oxygen via carbon monoxide (CO) and carbon dioxide (CO2). Traditionally, gas chromatography (GC) systems are utilized to quantify gases due to their ability to separate volatile compounds within the gas mixture, followed by detection of the isolated component of interest. Despite their capabilities, GCs require carrier gas consumables for their operational mobile phase and consequently are not often considered a flight-forward option. An alternative analytical technique known as mass spectrometry measures the presence of gas phase molecules without the need of carrier gas. Residual gas analyzers (RGAs), which are smaller and rugged mass spectrometers, are not traditionally used to quantify analytes as they serve more as a monitoring tool. However, proper application and calibration can result in a flight-forward analytical instrument capable of direct quantification of volatile compounds of interest. Kennedy Space Center's role for the JSC led CaRD project is to design a gas analyzer system to detect and quantify the CO and CO2 gases produced during the carbothermal reaction utilizing a commercial version of the Mass Spectrometer observing lunar operations (MSolo) instrument. MSolo is a modified commercial off-the-shelf RGA, which consists of quadrupole mass filter and space rated electronics. MSolo is currently a TRL 6 technology and manifested on the PRIME-1 and VIPER missions. This paper describes leveraging MSolo based instrumentation to develop a gas analysis system for the work outlined in the 2023 International Conference on Environmental Systems (ICES) paper number 313.

Ryan P Gott↗

Effect of rice husk ash-derived activator on the structural build-up of alkali activated materials

Highlights: • Filtered and unfiltered sodium silicate solutions were synthesised from RHA. • The dissolution efficiency of RHA in NaOH solution was 76.3%. • RHA-derived activator showed a structural build-up comparable to the commercial activator in AAM pastes. • The undissolved residues in the unfiltered RHA-derived activator increased the yield stress. • Comparable compressive strength and early age gel formation were observed from both commercial and RHA-derived solutions. Waste-derived sodium silicates (SSs) were synthesised from rice husk ash (RHA) to produce alkali activated materials (AAMs) using fly ash and slag as precursors. The effects of these SSs on the structural build-up were investigated by monitoring the evolution of the viscoelasticity, ultrasonic pulse waves, setting time, and infrared spectra. Two types of RHA derived SSs (filtered SS and unfiltered SS) were prepared to investigate the effect of the undissolved residue on the hardening process. The derived SS contained a large portion of monomer silicate species. The results showed that the structural build-up mechanism was comparable between the AAMs activated by the filtered SS and a commercial SS. The unfiltered SS increased the yield stress of the paste by up to 10 times. However, the residues in the unfiltered SS had insignificant impacts on the gel formation and compressive strength when using identical SiO{sub 2}/Na{sub 2}O and Na{sub 2}O/binder ratios in the mix design.

36 MATERIALS SCIENCE↗

The collaborative effects of intrinsic and extrinsic impurities in low RRR SRF cavities

The superconducting radio-frequency (SRF) community has shown that introducing certain impurities into high-purity niobium can improve quality factors and accelerating gradients. We question why some impurities improve RF performance while others hinder it. The purpose of this study is to characterize the impurity profile of niobium with a low residual resistance ratio (RRR) and correlate these impurities with the RF performance of low RRR cavities so that the mechanism of impurity-based improvements can be better understood and improved upon. The combination of RF testing and material analysis reveals a microscopic picture of why low RRR cavities experience low temperature-dependent BCS resistance behavior more prominently than their high RRR counterparts. We performed surface treatments, low temperature baking and nitrogen-doping, on low RRR cavities to evaluate how the intentional addition of oxygen and nitrogen to the RF layer further improves performance through changes in the mean free path and impurity profile. The results of this study have the potential to unlock a new understanding on SRF materials and enable the next generation of SRF surface treatments.

43 PARTICLE ACCELERATORS↗

The Collaborative Effects of Intrinsic and Extrinsic Impurities in Low RRR SRF Cavities

The superconducting radio-frequency (SRF) community has shown that introducing certain impurities into high-purity niobium can improve quality factors and accelerating gradients. We question why some impurities improve RF performance while others hinder it. The purpose of this study is to characterize the impurity profile of niobium with a low residual resistance ratio (RRR) and correlate these impurities with the RF performance of low RRR cavities so that the mechanism of impurity-based improvements can be better understood and improved upon. The combination of RF testing and material analysis reveals a microscopic picture of why low RRR cavities experience low temperature-dependent BCS resistance behavior more prominently than their high RRR counterparts. We performed surface treatments, low temperature baking and nitrogen-doping, on low RRR cavities to evaluate how the intentional addition of oxygen and nitrogen to the RF layer further improves performance through changes in the mean free path and impurity profile. The results of this study have the potential to unlock a new understanding on SRF materials and enable the next generation of SRF surface treatments.

43 PARTICLE ACCELERATORS↗

The effect of the isotopic composition of oxygen on the non-mass-dependent isotopic fractionation in the formation of ozone by discharge of O2

Isotopic fractionation processes in the formation of O3 by 20-kV 60-kHz discharge of O2 in a fused silica container at 77 K are investigated experimentally, with a focus on the effects of changes in the isotopic composition of O2, the O2 pressure, the degree of O3 formation, and the discharge geometry on the isotope abundances in the postdischarge O2 and O3. The results are presented in tables and graphs and compared with the predictions of theoretical models involving vibrational anharmonicity and symmetry effects on O3 predissociation. In experiments using O2 enriched with O-17 and O-18, the heavy isotope is enriched in the residual O2 and not in the O3, whereas the opposite is true when atmospheric isotope abundances are used. It is inferred that the predissociation rate depends on the symmetry and mass of the O3 molecules and not on self-shielding or symmetry effects in the reactant O2. The potential value of the present findings for studies of anomalous isotope abundances in meteorites is indicated.

Yang, Jongmann↗

Nepheline Crystallization Studies and the Structural Integrity of the Residual (SIR) - FY 2022 Study Glasses

In FY22 a set of 367 HLW glasses with compositions representative of the Waste Treatment and Immobilization Plant (WTP) processing region were aggregated and analyzed with the Structural Integrity of the Residual (SIR) model developed by the Savannah River National Laboratory (SRNL). An additional 12 glasses targeting 27 – 37 wt % alumina (in glass) and at waste loadings up to ~70 wt % were fabricated and added to the database. Collectively, this data set was analyzed with regression statistics and partitioning functions to reveal correlations between boron release and the chemical composition. FY22 results indicate that the SIR theoretical alkali-silicate precipitation term, Al 2 O 3 and B 2 O 3 concentration, and SIR computed non-bridging oxygen are significant predictors of chemical durability. A statistical model using the SIR parameters was shown to have greater overall accuracy than the nepheline discriminator model while maintaining false predictions of pour durability below 1%. In FY23, we will continue this work to focus on refining the SIR with glass compositions that challenge the SIR correlation terms, namely the theoretical crystalline precipitation and non-bridging oxygen terms.

36 MATERIALS SCIENCE↗

Influence of gasoline fuel formulation on lean autoignition in a mixed-mode-combustion (deflagration/autoignition) engine

We report stoichiometric spark-ignition engines suffer efficiency penalties due to throttling losses at low loads, a low specific-heat ratio of the stoichiometric working fluid, and limits on compression ratio due to end-gas autoignition leading to undesirable knocking. Mixed-Mode Combustion (MMC) mitigates these shortcomings by using a lean working fluid where a spark-initiated pilot-stabilized deflagrative flame front is followed by controlled end-gas autoignition. This MMC study investigates the effects of initial conditions (intake air temperature, intake pressure, equivalence ratio, and intake oxygen fraction) on autoignition tendency of four gasoline-range fuels with varying properties and composition. The use of fuels with varying octane sensitivity (S) allowed exploring the importance of low-temperature heat release in triggering autoignition. Fuels with high S were less reactive for conditions that promote low-temperature chemistry (operation at high intake air pressure or without N2 dilution). Conversely, an Alkylate fuel with low S showed a greater autoignition resistance at operating conditions that were unfavorable for low-temperature chemistry. Next, the effect of residual gas composition on autoignition tendency of fuels was examined with a chemical-kinetics model. Among the various molecules in the residual gas, nitric oxide (NO) enhanced the low-temperature chemistry and increased the autoignition tendency most significantly. The fuels’ autoignition response to increasing NO amount corroborates the experimental observations. Next, the sequential autoignition of the end-gas was assessed to be less impacted by thermal stratification because of lean mixtures showing relatively less low-temperature chemistry, when compared to stoichiometric mixtures. Next, the effect of changing equivalence ratio on the autoignition was found to be similar for all fuels, regardless of their S. With changing intake air temperature, the response of fuels’ autoignition tendency depended on the dilution level used. At high dilution (i.e. low intake [O2]), fuels’ reactivity increased with increasing intake air temperature. In contrast, for operation without dilution, the autoignition tendency of the low-S Alkylate fuel decreased with increasing intake air temperature, while that of high-S High Cycloalkane fuel still increased with increasing intake air temperature. In conclusion, conventional octane metrics (RON and MON) have utility in assessing the autoignition tendency under lean MMC operation. Moreover, the fuel requirements for MMC align with that of stoichiometric operation: i.e., high RON and high S fuels are desirable for stable non-knocking operation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Chlorophyllase from Arabidopsis thaliana Reveals an Emerging Model for Controlling Chlorophyll Hydrolysis

Chlorophyll (Chl) is one of Nature’s most complex pigments to biosynthesize and derivatize. This pigment is vital for survival and also paradoxically toxic if overproduced or released from a protective protein scaffold. Therefore, along with the mass production of Chl, organisms also invest in mechanisms to control its degradation and recycling. One important enzyme that is involved in these latter processes is chlorophyllase. This enzyme is employed by numerous photosynthetic organisms to hydrolyze the phytol tail of Chl. Although traditionally thought to catalyze the first step of Chl degradation, recent work suggests that chlorophyllase is instead employed during times of abiotic stress or conditions that produce reactive oxygen species. However, the molecular details regarding how chlorophyllases are regulated to function under such conditions remain enigmatic. Here, we investigate the Arabidopsis thaliana chlorophyllase isoform AtCLH2 using site-directed mutagenesis, mass spectrometry, dynamic light scattering, size-exclusion multiangle light scattering, and both steady-state enzyme kinetic and thermal stability measurements. Through these experiments, we show that AtCLH2 exists as a monomer in solution and contains two disulfide bonds. One disulfide bond putatively maps to the active site, whereas the other links two N-terminal Cys residues together. These disulfide bonds are cleaved by chemical or chemical and protein-based reductants, respectively, and are integral to maintaining the activity, stability, and substrate scope of the enzyme. This work suggests that Cys residue oxidation in chlorophyllases is an emerging regulatory strategy for controlling the hydrolysis of Chl pigments.

59 BASIC BIOLOGICAL SCIENCES↗

Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000

Pseudomonas syringaepv.tomatoDC3000 (PstDC3000) is a plant pathogenic bacterium that possesses complicated motility regulation pathways including a typical chemotaxis system. A significant portion of our understanding about the genes functioning inPstDC3000 motility is based on comparison to other bacteria. This leaves uncertainty about whether gene functions are conserved, especially since specific regulatory modules can have opposite functions in sets ofPseudomonas. In this study, we used a competitive selection to enrich for mutants with altered swimming motility and used random barcode transposon-site sequencing (RB-TnSeq) to identify genes with significant roles in swimming motility. Besides many of the known or predicted chemotaxis and motility genes, our method identified PSPTO_0406 (dipA), PSPTO_1042 (chrR) and PSPTO_4229 (hypothetical protein) as novel motility regulators. PSPTO_0406 is a homolog ofdipA, a known cyclic di-GMP degrading enzyme inP. aeruginosa. PSPTO_1042 is part of an extracytoplasmic sensing system that controls gene expression in response to reactive oxygen species, suggesting that PSPTO_1042 may function as part of a mechanism that enablesPstDC3000 to alter motility when encountering oxidative stressors. PSPTO_4229 encodes a protein containing an HD-related output domain (HDOD), but with no previously identified functions. We found that deletion and overexpression of PSPTO_4229 both reduce swimming motility, suggesting that its function is sensitive to expression level. We used the overexpression phenotype to screen for nonsense and missense mutants of PSPTO_4229 that no longer reduce swimming motility and found a pair of conserved arginine residues that are necessary for motility suppression. Together these results provide a global perspective on regulatory and structural genes controlling flagellar motility inPstDC3000.

Microbiology↗

Low thrust chemical orbit to orbit propulsion system propellant management study

Low thrust chemical propulsion systems were sized for transfer of large space systems from LEO to GEO. The influence of propellant combination, tankage and insulation requirements, and propellant management techniques on the LTPS mass and volume were studied. Liquid oxygen combined with hydrogen, methane or kerosene were the propellant combinations. Thrust levels of 445, 2230, and 4450 N were combined with 1, 4 and 8 perigee burn strategies. This matrix of systems was evaluated using multilayer insulation and spray-on-foam insulation systems. Various combinations of toroidal, cylindrical with ellipsoidal domes, and ellipsoidal tank shapes were investigated. Results indicate that low thrust (445 N) and single perigee burn approaches are considerably less efficient than the higher thrust level and multiple burn strategies. A modified propellant settling approach minimized propellant residuals and decreased system complexity, in addition, the toroid/ellipsoidal tank combination was predicted to be shortest.

Dergance, R. H.↗

Accuracy Quantification of the Loci-CHEM Code for Chamber Wall Heat Transfer in a GO2/GH2 Single Element Injector Model Problem

A robust rocket engine combustor design and development process must include tools which can accurately predict the multi-dimensional thermal environments imposed on solid surfaces by the hot combustion products. Currently, empirical methods used in the design process are typically one dimensional and do not adequately account for the heat flux rise rate in the near-injector region of the chamber. Computational Fluid Dynamics holds promise to meet the design tool requirement, but requires accuracy quantification, or validation, before it can be confidently applied in the design process. This effort presents the beginning of such a validation process for the Loci-CHEM CFD code. The model problem examined here is a gaseous oxygen (GO2)/gaseous hydrogen (GH2) shear coaxial single element injector operating at a chamber pressure of 5.42 MPa. The GO2/GH2 propellant combination in this geometry represents one the simplest rocket model problems and is thus foundational to subsequent validation efforts for more complex injectors. Multiple steady state solutions have been produced with Loci-CHEM employing different hybrid grids and two-equation turbulence models. Iterative convergence for each solution is demonstrated via mass conservation, flow variable monitoring at discrete flow field locations as a function of solution iteration and overall residual performance. A baseline hybrid was used and then locally refined to demonstrate grid convergence. Solutions were obtained with three variations of the k-omega turbulence model.

West, Jeff↗

Accuracy Quantification of the Loci-CHEM Code for Chamber Wall Heat Fluxes in a G02/GH2 Single Element Injector Model Problem

A robust rocket engine combustor design and development process must include tools which can accurately predict the multi-dimensional thermal environments imposed on solid surfaces by the hot combustion products. Currently, empirical methods used in the design process are typically one dimensional and do not adequately account for the heat flux rise rate in the near-injector region of the chamber. Computational Fluid Dynamics holds promise to meet the design tool requirement, but requires accuracy quantification, or validation, before it can be confidently applied in the design process. This effort presents the beginning of such a validation process for the Loci- CHEM CPD code. The model problem examined here is a gaseous oxygen (GO2)/gaseous hydrogen (GH2) shear coaxial single element injector operating at a chamber pressure of 5.42 MPa. The GO2/GH2 propellant combination in this geometry represents one the simplest rocket model problems and is thus foundational to subsequent validation efforts for more complex injectors. Multiple steady state solutions have been produced with Loci-CHEM employing different hybrid grids and two-equation turbulence models. Iterative convergence for each solution is demonstrated via mass conservation, flow variable monitoring at discrete flow field locations as a function of solution iteration and overall residual performance. A baseline hybrid grid was used and then locally refined to demonstrate grid convergence. Solutions were also obtained with three variations of the k-omega turbulence model.

West, Jeff↗

Revealing the atomic and electronic mechanism of human manganese superoxide dismutase product inhibition

Human manganese superoxide dismutase (MnSOD) is a crucial oxidoreductase that maintains the vitality of mitochondria by converting superoxide (O 2 •– ) to molecular oxygen (O 2 ) and hydrogen peroxide (H 2 O 2 ) with proton-coupled electron transfers (PCETs). Human MnSOD has evolved to be highly product inhibited to limit the formation of H 2 O 2 , a freely diffusible oxidant and signaling molecule. The product-inhibited complex is thought to be composed of a peroxide (O 2 2– ) or hydroperoxide (HO 2 – ) species bound to Mn ion and formed from an unknown PCET mechanism. PCET mechanisms of proteins are typically not known due to difficulties in detecting the protonation states of specific residues that coincide with the electronic state of the redox center. To shed light on the mechanism, we combine neutron diffraction and X-ray absorption spectroscopy of the product-bound, trivalent, and divalent states of the enzyme to reveal the positions of all the atoms, including hydrogen, and the electronic configuration of the metal ion. The data identifies the product-inhibited complex, and a PCET mechanism of inhibition is constructed.

59 BASIC BIOLOGICAL SCIENCES↗

Early photosynthetic microorganisms and environmental evolution

Microfossils which are preserved as shrivelled kerogenous residues provide little information about cellular organization and almost none about the metabolic properties of the organisms. The distinction between prokaryotic vs eukaryotic, and phototrophic vs chemo- and organotrophic fossil microorganisms rests entirely on morphological comparisons with recent counterparts. The residual nature of the microbial fossil record promotes the conclusion that it must be biased toward (a) most abundant organisms, (b) those most resistant to degradation, and (c) those inhabiting environments with high preservation potential e.g., stromatolites. These criteria support the cyanophyte identity of most Precambrian microbial fossils on the following grounds: (1) as primary producers they dominate prokaryotic communities in modern extreme environments, e.g., intertidal zone; (2) several morphological counterparts of modern cyanophytes and microbial fossils have been established based on structure, cell division patterns and degradation sequences. The impact of anaerobic and oxygenic microbial photosynthesis on the evolution of Precambrian environments is discussed.

Golubic, S.↗

Launch Complex 39B, SWMU 009 Air Sparging System Expansion Implementation Work Plan Kennedy Space Center, Florida

This Implementation Work Plan (IWP) for expansion of the Air Sparging (AS) system at Launch Complex 39B (LC39B), presents the design and coordination specifics to expand the existing AS Interim Measure (IM) at LC39B located at Kennedy Space Center (KSC), Florida. The existing AS system at LC39B was installed in 2017 and included 279 AS wells. LC39B has been designated as Solid Waste Management Unit (SWMU) 009 under the KSC Resource Conservation and Recovery Act (RCRA) Corrective Action Program. Expansion of the AS IM will include installation of an additional five AS wells to depths of approximately 13 and 17 feet below land surface (bls) to remediate contaminated groundwater identified northwest of the launch pad, near the liquid oxygen (LOX) tank. The expansion will target concentrations of cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC) greater than Florida Department of Environmental Protection (FDEP) Natural Attenuation Default Concentrations (NADCs) that were identified during direct push technology (DPT) sampling events between 2019-2022 in shallow intervals between 8 and 16 feet bls. The objective for this expansion is to remediate contaminated groundwater within the treatment zone to below NADCs to support transition to monitored natural attenuation (MNA) through a long-term monitoring (LTM) program. This expansion will treat approximately 0.07-acres of residual groundwater contamination. The IWP design for this expansion will utilize the existing AS system trailer that is already onsite and operating. The five new AS expansion wells will be integrated into the existing Zone Z4 distribution manifold to deliver compressed air to the new AS wells. To deliver airflow, three existing lateral lines will be split in the field at a lateral split junction point (using 2-way or 3-way splits). Each split lateral will supply air to AS wells at the same depth interval to minimize preferential pathways. The trenching layout for the new AS wells was developed to minimize ground disturbance due to the amount of utilities in the area. One new monitoring well will also be installed and added to LC39B’s performance monitoring program to evaluate the effectiveness of the AS system expansion. This well will be sampled on a semi-annual basis for volatile organic compounds, with results evaluated to determine if system adjustments may be required. The main components of this IWP were presented to the KSC Remediation Team (KSCRT) at the October 2022 Meeting (Meeting 2210-M05). DPT delineation data from the LOX area were also presented, which included some inferred boundaries due to physical restrictions of the LOX tank and associated concrete pad and infrastructure. Planned monitoring well sampling in this area will help determine if back-diffusion may be occurring from beneath the pad. Additionally, based on potential petroleum odors and staining identified at two DPT locations during the January 2022 DPT event in the 10-16 feet bls range, a KSCRT action item was assigned during the October 2022 meeting to collect soil samples for petroleum compounds if similar odors or staining are identified during IM implementation. Construction completion and performance monitoring will be incorporated into LC39B’s groundwater monitoring reports, which are currently submitted on an annual basis to FDEP to document operation and maintenance activities and results from the AS IM.

Deborah M. Wilson↗

Air Classification of Forestry Residues for Fast Pyrolysis

Understanding critical biomass attributes through efficient fractionation is crucial for advancing sustainable pyrolysis for renewable energy and chemical production. This study investigates the intricate relationship between biomass preprocessing and pyrolysis product yields, employing the air classification technique for the treatment of loblolly pine residues with varying moisture content. A comprehensive exploration of the physicochemical properties of air-classified loblolly pine informs a sophisticated pyrolysis simulation model. Given the complex and multifaceted nature of biomass pyrolysis, operating across diverse temporal and spatial scales, a pyrolysis kinetics-based CFD–DEM simulation method is employed to predict product yields. Results showed that the elevated moisture content amplifies particle adhesiveness, necessitating augmented air velocities for effective separation, thereby influencing the efficiency of the separation process. While carbon and hydrogen contents exhibit relative stability across diverse moisture contents and blower frequencies, the oxygen content undergoes noticeable changes. For example, the oxygen contents were measured as 29.2 and 38.6 wt% in the light fraction of 30% moisture content sample at blower frequencies of 10 and 20 Hz, respectively. An intriguing finding emerges from pyrolysis simulation, indicating that a lower blower frequency in air classification moderately enhances bio-oil yield and significantly improves its quality, particularly in terms of water content. For instance, the water content in the bio-oil was about 1.5% and 10% in the heavy and light fractions, respectively from 10% moisture sample under 15 Hz blower frequency.

09 - BIOMASS FUELS↗

Structure of active methyl-CoM reductase, Earth’s main methane producer

Summary Our work reveals the structure of the active state of Methyl-Coenzyme M Reductase (MCR), the key and rate-limiting enzyme in biological methane formation. We find large differences between the active Ni(I) and inactive Ni(II) proteins and provide insight into how nature makes and breaks the C-H bond of methane. The Ni(II)-F430 center in inactive MCR contains four planar nitrogen ligands, a lower axial glutamine oxo, and an upper axial thiolate. The Ni(I)-enzyme replaces the axial ligands with a single water. The one-electron redox change results in movement of the Ni ion and upward swing of the β-lactam ring in the tetrapyrrole coupled to a domino-like protein quake through second sphere residues, inter-subunit interactions, a substrate tunnel, affecting even the dimensions of the unit cell. These structural changes lead Ni(I)-MCR to release a charge clamp that, in the Ni(II) state, locks down substrate Coenzyme B. Determining the Ni(I)-MCR structure required development of rigorous anaerobic crystallographic techniques. Validation of the MCR redox state was accomplished by in-line and parallel spectroscopic and unit cell analyses. This structure has large implications for developing technologies to limit methane emissions and efficiently produce biofuels. Methodology described here will enhance structural biology for other oxygen-sensitive enzymes.

09 BIOMASS FUELS↗