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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.

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At least 145 records · Page 8
All sourcesNASA NTRSDOE OSTI
DOE OSTI2020

Atomistic Characterization of Gramicidin Channel Formation

We investigated gramicidin A (gA) subunit dimerization in lipid bilayers using microsecond-long replica-exchange umbrella sampling simulations, millisecond-long unbiased molecular dynamics simulations, and machine learning. Our simulations led to a dimer structure that is indistinguishable from the experimentally determined gA channel structures, with the two gA subunits joined by six hydrogen bonds (6HB). The simulations also uncovered two additional dimer structures, with different gA–gA stacking orientations that were stabilized by four or two hydrogen bonds (4HB or 2HB). When examining the temporal evolution of the dimerization, we found that two bilayer-inserted gA subunits can form the 6HB dimer directly, with no discernible intermediate states, as well as through paths that involve the 2HB and 4HB dimers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

Characterization of Heparin’s Conformational Ensemble by Molecular Dynamics Simulations and Nuclear Magnetic Resonance Spectroscopy

Heparin is a highly charged, polysulfated polysaccharide and serves as an anticoagulant. Heparin binds to multiple proteins throughout the body, suggesting a large range of potential therapeutic applications. Although its function has been characterized in multiple physiological contexts, heparin’s solution conformational dynamics and structure–function relationships are not fully understood. Molecular dynamics (MD) simulations facilitate the analysis of a molecule’s underlying conformational ensemble, which then provides important information necessary for understanding structure–function relationships. However, for MD simulations to afford meaningful results, they must both provide adequate sampling and accurately represent the energy properties of a molecule. The aim of this study is to compare heparin’s conformational ensemble using two well-developed force fields for carbohydrates, known as GLYCAM06 and CHARMM36, using replica exchange molecular dynamics (REMD) simulations, and to validate these results with NMR experiments. In this work, the anticoagulant sequence, an ultra-low-molecular-weight heparin, known as Arixtra (fondaparinux, sodium), was simulated with both parameter sets. The results suggest that GLYCAM06 matches experimental nuclear magnetic resonance three-bond J-coupling values measured for Arixtra better than CHARMM36. In addition, NOESY and ROESY experiments suggest that Arixtra is very flexible in the sub-millisecond time scale and does not adopt a unique structure at 25 C. Moreover, GLYCAM06 affords a much more dynamic conformational ensemble for Arixtra than CHARMM36.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Pulse Radiolysis Investigation of Radicals Derived from Water-Soluble Cyanine Dyes: Implications for Super-resolution Microscopy

Light-induced blinking, an inherent feature of many forms of super-resolution microscopy, has been linked to transient reduction of the fluorescent cyanine dye used as an imaging agent. There is, however, only scant literature information related to one-electron reduced cyanine dyes, especially in an aqueous environment. Here, we examine a small series of cyanine dyes, possessing disparate π-conjugation lengths, under selective reducing or oxidizing conditions. The experiment allows recording of both differential absorption spectra and decay kinetics of the resultant one-electron reduced or oxidized transient species in water. Relative to the ground state, absorption transitions for the various radicals are weak and somewhat broadened but do allow correlation with the π-conjugation length. In all cases, absorption maxima lie to the blue of the main ground-state transition. Under anaerobic conditions, the transient species decay on the microsecond to millisecond time scale, with the mean lifetime depending on molecular structure, radiation dose, and dye concentration. The experimental absorption spectra recorded for the one-electron reduced radicals and the presumed dimer cation radical compare well to spectra obtained from time-dependent density functional theory calculations. Furthermore, the results allow conclusions to be drawn regarding the plausibility of the reduced species being responsible for light-induced blinking in direct stochastic optical reconstruction microscopy.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

Microsized Pore Structure Determination in EPDM Rubbers Using High-Pressure 129 Xe NMR Techniques

Microsized pore parameters, such as pore size and distance between pores in a series of model EPDM rubbers, were determined in situ under the pressure of 500 psi using 129 Xe nuclear magnetic resonance (NMR) techniques: spin–lattice (T 1 ) and spin–spin (T 2 ) relaxation measurements, pulsed-field gradient (PFG) NMR, and two-dimensional exchange spectroscopy (2D EXSY). The T 1 /T 2 (>>1) ratio for the xenon confined in the pores is larger than that for nonconfined free xenon. This suggests that almost the entire pore surface interacts with xenon atoms like a closed pore. While these pores still connect each other through very narrow diffusion/exchange channels, it is possible to observe the echo decay in PFG-NMR and cross-peaks in 2D EXSY. The results show that both diffusion (D pore ≈ 2.1 × 10 –10 m 2 /s) and exchange (exchange rate, τ exch = a few tens of milliseconds) of xenon between a pore within the material and outer surface are prolonged. The exchange distances (l), which correspond to the xenon gas penetration depth, were estimated to be 70–100 μm based on the measured diffusion coefficients and exchange rate (1/τ exch ). NMR diffraction analysis reveals that pore size (a) and pore distance (b) are on the order of magnitude of micrometers and tens of micrometers, while the diffusion coefficients of xenon gas in the diffusion channels (D eff ) are about 10 –8 m 2 /s. Overall, this study suggests that the pores with a few micrometers connected through very narrow flowing channels with the length of several tens of micrometers are developed 70 to 100 μm below the rubber surface. Furthermore, the overall steady-state diffusion of xenon is slower, approximately 2 orders of magnitudes, than the diffusion in the channel between the pores. Finally, the pore and exchange distances correlated with the composition of rubbers showed that the properties of EPDM rubber as a high-pressure gas barrier could be improved by reducing the size of cracks and the depth of gas penetration by the addition of both carbon black and silica fillers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Electron Transfer Going the Distance: Mn-Doped ZnSe as a Model Photocatalytic System

In this work, time-resolved photoluminescence and transient absorption spectroscopy are used to examine the influence that long excited state lifetimes of Mn-doped nanocrystals have on the mechanisms of photo-induced electron transfer (PET) in photocatalytic systems. Mn-doped ZnSe nanocrystals can undergo PET over their nearly millisecond excited state lifetime, which enables electron transfer to viologens in solution over large encounter distances in comparison to molecular length scales. The long excited state lifetimes also enable diffusion of the excited nanocrystals over hundreds of nanometers in solution, allowing them to react with molecular species at nanomolar concentrations. The ability to capture and sustain optical energy in spin-forbidden transitions among crystal field states of the Mn ions opens opportunities to explore the influence that long excited state lifetimes have on photocatalytic reaction mechanisms involving molecular species such as CO 2 that can be difficult to concentrate or attach to nanocrystal surfaces.

14 SOLAR ENERGY↗
DOE OSTI2025

Strategic Lifetime Tuning of Visible-Light Absorbing Two-Coordinate Metal Complexes

This paper highlights how the singlet and triplet amide ligand centered ( 1,3 LC) and interligand charge transfer ( 1,3 ICT) states in (carbene)M(amide), M = Cu, Au (cMa) complexes influence the excited state properties when the triplet states are close in energy. To that end we prepared a set of five cMa complexes, in which the amide (i.e., 5H-benzo[b]carbazole, H-BnCz) was kept constant giving a 3 LC energy of ca. 2.15 eV. Four different carbene ligands were selected to develop M BnCz Carbene complexes which have energies for the ICT state that vary from being either markedly higher, lower, or close to the energy of the 3 LC state on the amide ligand. Steady-state and time-resolved spectroscopic studies show that the emission spectrum of the cMa complex mirrors the phosphorescence of the parent H-BnCz amide and has a luminescence decay lifetime in the millisecond regime when the lowest energy excited state is 3 LC. When the lowest energy states are 1,3 ICT in nature, the emission band is broad and featureless, giving a lifetime in the nanosecond regime. As the 3 LC and 1,3 ICT states have comparable energies, dynamic equilibrium between the states is observed with the luminescence consisting of a mixture of 3 LC and 1,3 ICT transitions. The measured lifetime of this equilibrating system is between 45 and 350 μs depending on the solvent, for both copper- and gold-based cMa complexes. Furthermore, these complexes demonstrate the ability to “park” the excited state population in the 3 LC state, allowing it to act as a reservoir for thermally activated emission, while still maintaining a very rapid equilibrium between LC and ICT states.

14 SOLAR ENERGY↗
DOE OSTI2022

Excited State Exchange Control of Photoinduced Electron Spin Polarization in Electronic Ground States

Ground-state electron spin polarization (ESP) is generated in radical elaborated (bpy)Pt(CAT-NN) and (bpy)Pt(CAT- p -Me 2 PhMe 2 -NN) (bpy = 5,5'-di- tert -butyl-2,2'-bipyridine, CAT = 3- tert -butylcatecholate, p -Ph = para -phenylene, NN = nitronylnitroxide). Photoexcitation produces an exchange-coupled, three-spin, charge-separated doublet 2 S 1 (S = chromophore excited spin singlet configuration) excited state that rapidly decays to a 2 T 1 (T = chromophore excited spin triplet configuration) excited state. The SQ-bridge-NN bond torsions affect the magnitude of the excited state exchange interaction ( J SQ-NN ), which determines the 2 T 1 – 4 T 1 energy gap. Ground state ESP is dependent on the magnitude of J SQ-NN , and we postulate that this results from differences in 2 T 1 and 4 T 1 state mixing. Mechanisms that lead to the rapid transfer of the excited state ESP to the ground state are discussed. Although subnanosecond 2 T 1 state lifetimes are measured optically in solution, the ground state ESP decays very slowly at 20 K and is observable for more than a millisecond.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2020

Effects of Photonic Curing Processing Conditions on MAPbI 3 Film Properties and Solar Cell Performance

Thermal annealing is the most used postdeposition materials processing method in laboratory research, but due to its slow speed and high energy cost, it is not compatible with the upscaling and commercialization of perovskite solar cell (PSC) manufacturing. Here, we adapt photonic curing (PC), which uses millisecond light pulses to deliver energy to the sample, to replace thermal annealing for crystallization of methylammonium lead iodide (MAPbI 3 ) films and rapid fabrication of PSCs. We study how PC conditions affect the outcome of MAPbI 3 conversion from the precursor to the crystalline perovskite phase by evaluating the films’ optical, crystalline, and morphological properties, as well as PSC performance. The results are understood using simulated film temperature profiles. We show that MAPbI 3 is readily converted under a wide range of PC conditions. While previous reports all used short pulses (<3 ms), we find that longer pulses produce more dense films and higher-performing PSCs. We achieve a champion power conversion efficiency in a PC-processed MAPbI 3 PSC of 11.26% under forward scan and 10.34% under reverse scan, with the processing time for the MAPbI 3 layer reduced by 30,000-fold, from 10 min to 20 ms. Using a 6 in. lamp, spatial uniformity tests show a cross-web efficiency variation of 5%. Our results indicate that using longer pulse lengths, >10 ms, is the best PC strategy for perovskite conversion, and PC is a promising annealing method for large-area, high-throughput PSC manufacturing.

14 SOLAR ENERGY↗
DOE OSTI2020

Intense Pulse Light Annealing of Perovskite Photovoltaics Using Gradient Flashes

Perovskite solar cells (PSCs) have been fabricated through high-speed and low-cost depositions but often require long annealing. Intense pulse light (IPL) can anneal thin films in seconds after deposition by inducing very high temperatures lasting for milliseconds, and multiple flashes can be used to tune the temperature profile. In this study, a gradient flash annealing (GFA) approach is introduced and compared to uniform flash annealing (UFA) by investigating the crystallinity, morphology, and phase evolution of the CH 3 NH 3 PbI 3 perovskite films and their impact on PSC performance. Unlike UFA, low-intensity pulsed irradiation during the pre-annealing stage of GFA played a significant role on enhancing the PSC performance by forming pure-phase CH 3 NH 3 PbI 3 perovskite thin films with superior morphology and high crystallinity. To understand the kinetics, a transient thermal simulation using ANSYS was developed and confirmed with an experimental setup. The results combined with microscopy and spectrophotometry were used to visualize how duration, delay time, photon flux, and flash count parameters participated in crystallization, phase, and morphology evolution. The IPL annealing induced rapid surface temperature increase, reaching as high as 800 °C, while produced well-developed and bound perovskite grains without any surface defects in an uncontrolled ambient environment with high humidity (>60%). The study resulted in PSCs with maximum efficiency and fill factor of 9.27 and 69.92% for the UFA and 11.75 and 68% when annealed through the GFA approach, respectively. Lastly, this work utilized IPL as the sole thermal source for post-deposition annealing to rapidly fabricate efficient PSCs in only 10 s, which opens the pathway for high-speed, low-cost, and large-scale automated fabrication of perovskite photovoltaics and semiconductors.

14 SOLAR ENERGY↗
DOE OSTI2025

Photoabsorption and Stability in Triple-Cation Perovskites Influenced by Interfacial Engineering of the Collector

Charge carrier dynamics in three-dimensional (3D) perovskites is critical to understand for enhancing device performance since the photoabsorption mechanism in perovskites influences key functional devices such as photodetectors and solar cells. Temperature-dependent optoelectronic transport measurements were conducted on our triple cation formulation for the first time from 4 K to 300 K to investigate the role of an interfacial Ti underlayer, beneath the conventional Au collector electrode. The photocurrent was 10X larger with the use of a Ti interfacial layer compared to only Au, where device measurements were made using a broadband white light source at room temperature. As temperature increased from 4 K, the photocurrent increased in both cases, consistent with the semiconducting nature of the triple-cation absorber. Besides computing the responsivity as a function of power and temperature, time-domain measurements with ON/OFF pulses of incident white light, showed the switching time constants to be in the tens to few hundred milliseconds range, and largely temperature-invariant for the two contacts examined. Finally, we constructed solar cells with the same triple cation absorber, in an n-i-p architecture with a Spiro-OMeTAD hole transport layer, but the collector was composed of both types of contacts. Exposing our devices to moisture-rich conditions of up to 70% relative humidity showed the Au/Ti contacted devices to be more robust. Here, our experimental results demonstrate that the addition of a Ti interlayer improves collector efficiency through the photoabsorption process while also potentially stabilizing the solar cells, compared to the bare Au, in moisture-rich environments

optoelectronics↗
DOE OSTI2020

Visualizing Charge Transport and Nanoscale Electrochemistry by Hyperspectral Kelvin Probe Force Microscopy

Charge-transport and electrochemical processes are heavily influenced by the local microstructure. Kelvin probe force microscopy (KPFM) is a widely used technique to map electrochemical potentials at the nanometer scale; however, it offers little information on local charge dynamics. In this study, we implement a hyperspectral KPFM approach for spatially mapping bias-dependent charge dynamics in timescales ranging from the sub-millisecond to the second regime. As a proof of principle, we investigate the role mobile surface charges play in a three-unit-cell LaAlO 3 /SrTiO 3 oxide heterostructure. We explore machine learning approaches to assist with visualization, pattern recognition, and interpretation of the information-rich data sets. Linear unmixing methods reveal hidden bias-dependent interfacial processes, most likely water splitting, which are essentially unnoticed by functional fitting of the dynamic response alone. Hyperspectral KPFM will be beneficial for investigating nanoscale charge transport and local reactivity in systems involving a possible combination of electronic, ionic, and electrochemical phenomena.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Electrolyte Immersion Increases Photoconductivity in a Model Polymer Photocathode

Immersing polymer solar cells in aqueous electrolyte for photoelectrochemical (PEC) hydrogen production is likely to cause photophysical changes that could present both challenges and opportunities for engineering functional and durable devices. Herein we study the bulk heterojunction blend poly(4,8-bis(5-(2-ethylhexyl)thiophen-2-yl)benzo[1,2-b:4,5-b']dithiophene-2,6-diyl)-alt-(2-(((2-ethylhexyl)oxy)carbonyl)-3-fluorothieno[3,4-b]thiophene-4,6-diyl):poly(N,N'-di(2-octyldodecyl)naphthalene-1,8:4,5-bis(dicarboximide)-2,6-diyl)-alt-(2,2-bithiophene-5,5'-diyl) (PTB7-Th:N2200) excited-state dynamics in electrolyte from femtosecond to millisecond time scales using pump-probe microwave conductivity and absorption spectroscopy. While the blend swells very little, electrolyte exposure increases the microwave-frequency mobility and possibly the yield of photogenerated charges while also decreasing crystallinity. These results indicate an enhancement in key performance metrics, implying that any limitations on the performance of PEC test devices do not arise from active layer-electrolyte interactions. For the PTB7-Th:N2200 blend or similar photocathode systems, our results indicate that improving the interfacial kinetics and/or the carrier lifetime should be prioritized, not protecting the active layer from the electrolyte. Since this observation may not be universal to all polymer systems, future research should focus on identifying their limiting photophysical processes. electrolyte.

14 SOLAR ENERGY↗
DOE OSTI2021

Ultrafast and Controllable Phase Evolution by Flash Joule Heating

Flash Joule heating (FJH), an advanced material synthesis technique, has been used for the production of high-quality carbon materials. Direct current discharge through the precursors by large capacitors has successfully converted carbon-based starting materials into bulk quantities of turbostratic graphene by the FJH process. However, the formation of other carbon allotropes, such as nanodiamonds and concentric carbon materials, as well as the covalent functionalization of different carbon allotropes by the FJH process, remains challenging. Here, we report the solvent-free FJH synthesis of three different fluorinated carbon allotropes: fluorinated nanodiamonds, fluorinated turbostratic graphene and fluorinated concentric carbon. This is done by millisecond flashing of organic fluorine compounds and fluoride precursors. Spectroscopic analysis confirms the modification of the electronic states and the existence of various short-range and long-range orders in the different fluorinated carbon allotropes. As a result, the flash time-dependent relationship is further demonstrated to control the phase evolution and product compositions.

01 COAL, LIGNITE, AND PEAT↗
DOE OSTI2022

Heteroatom-Doped Flash Graphene

Heteroatom doping can effectively tailor the local structures and electronic states of intrinsic two-dimensional materials, and endow them with modified optical, electrical, and mechanical properties. Recent studies have shown the feasibility of preparing doped graphene from graphene oxide and its derivatives via some post-treatments, including solid-state and solvothermal methods, but they require reactive and harsh reagents. However, direct synthesis of various heteroatom-doped graphene in larger quantities and high purity through bottom-up methods remains challenging. Here, we report catalyst-free and solvent-free direct synthesis of graphene doped with various heteroatoms in bulk via flash Joule heating (FJH). Seven types of heteroatom-doped flash graphene (FG) are synthesized through millisecond flashing, including single-element-doped FG (boron, nitrogen, oxygen, phosphorus, sulfur), two-element-co-doped FG (boron and nitrogen), as well as three-element-co-doped FG (boron, nitrogen, and sulfur). Here, a variety of low-cost dopants, such as elements, oxides, and organic compounds are used. The graphene quality of heteroatom-doped FG is high, and similar to intrinsic FG, the material exhibits turbostraticity, increased interlayer spacing, and superior dispersibility. Electrochemical oxygen reduction reaction of different heteroatom-doped FG is tested, and sulfur-doped FG shows the best performance. Lithium metal battery tests demonstrate that nitrogen-doped FG exhibits a smaller nucleation overpotential compared to Cu or undoped FG. The electrical energy cost for the synthesis of heteroatom-doped FG synthesis is only 1.2 to 10.7 kJ g –1 , which could render the FJH method suitable for low-cost mass production of heteroatom-doped graphene.

01 COAL, LIGNITE, AND PEAT↗
DOE OSTI2023

Ultrafast Switching of Interfacial Thermal Conductance

Dynamical control of thermal transport at the nanoscale provides a time-domain strategy for optimizing thermal management in nanoelectronics, magnetic devices, and thermoelectric devices. However, the rate of change available for thermal switches and regulators is limited to millisecond time scales, calling for a faster modulation speed. Here, in this work, time-resolved X-ray diffraction measurements and thermal transport modeling reveal an ultrafast modulation of the interfacial thermal conductance of an FeRh/MgO heterostructure as a result of a structural phase transition driven by optical excitation. Within 90 ps after optical excitation, the interfacial thermal conductance is reduced by a factor of 5 and lasts for a few nanoseconds, in comparison to the value at the equilibrium FeRh/MgO interface. The experimental results combined with thermal transport calculations suggest that the reduced interfacial thermal conductance results from enhanced phonon scattering at the interface where the lattice experiences transient in-plane biaxial stress due to the structural phase transition of FeRh. Our results suggest that optically driven phase transitions can be utilized for ultrafast nanoscale thermal switches for device application.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Ultrafast Photoflash Synthesis of High-Entropy Oxide Nanoparticles

High-entropy metal oxides (HEOs) have recently received growing attention for broad energy conversion and storage applications due to their tunable properties. HEOs typically involve the combination of multiple metal cations in a single oxide lattice, thus bringing distinctive structures, controllable elemental composition, and tunable functional properties. Many synthesis methods for HEOs have been reported, such as solid-state reactions and carbon thermal shock methods. These methods frequently are energy-intensive or require relatively expensive heating equipment. In this work, we report an ultrafast photoflash synthesis method for HEO nanoparticles on diverse substrates. The energy input is provided by a commercial Xe photoflash unit, which triggers exothermic reactions to convert metal salt precursors to HEO nanoparticles within tens of milliseconds. The formation of HEO nanoparticles is attributed to the ultrafast heating (∼106 K/s) and cooling (∼105 K/s) rates of the photoflash and overall high temperature (>1000 K) during the ultrafast synthesis process. When the synthesized CoNiFeCrMn oxide (HEO) is tested as an oxygen evolution reaction electrocatalyst, it shows similar activity to similar materials prepared by other methods. We believe this photoflash synthesis provides a simple method for many others to synthesize diverse HEOs and explore their properties and potential applications.

exothermic reaction synthesis↗
DOE OSTI2020

Strong Electronic and Magnetic Coupling in M 4 (M = Ni, Cu) Clusters via Direct Orbital Interactions between Low-Coordinate Metal Centers

In this report we present an extensive study of tetranuclear transition-metal cluster compounds M 4 (NP t Bu 3 ) 4 and [M 4 (NP t Bu 3 ) 4 ][B(C 6 F 5 ) 4 ] (M = Ni, Cu; t Bu = tert -butyl), which feature low-coordinate metal centers and direct metal-metal orbital overlap. X-ray diffraction, electrochemical, magnetic, spectroscopic, and computational analysis elucidate the nature of the bonding interactions in these clusters and the impact of these interactions on the electronic and magnetic properties. Direct orbital overlap results in strongly coupled, large-spin ground states in the [Ni 4 (NP t Bu 3 ) 4 ] +/0 clusters and fully delocalized, spin-correlated electrons. Correlated electronic structure calculations confirm the presence of ferromagnetic ground states that arise from direct exchange between magnetic orbitals, and, in the case of the neutral cluster, itinerant electron magnetism similar to that in metallic ferromagnets. The cationic nickel cluster also possesses large magnetic anisotropy exemplified by a large, positive axial zero-field splitting parameter of D = +7.95 or +9.2 cm -1 , as determined by magnetometry or electron paramagnetic resonance spectroscopy, respectively. The [Ni 4 (NP t Bu 3 ) 4 ] + cluster is also the first molecule with easy-plane magnetic anisotropy to exhibit zero-field slow magnetic relaxation, and under a small applied field, it exhibits relaxation exclusively through an Orbach mechanism with a spin relaxation barrier of 16 cm -1 . The S = 1 / 2 complex [Cu 4 (NP t Bu 3 ) 4 ] + exhibits slow magnetic relaxation via a Raman process on the millisecond time scale, supporting the presence of slow relaxation via an Orbach process in the nickel analogue. Overall, this work highlights the unique electronic and magnetic properties that can be realized in metal clusters featuring direct metal-metal orbital interactions between low-coordinate metal centers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2023

In Situ Structural Observation of a Substrate- and Peroxide-Bound High-Spin Ferric-Hydroperoxo Intermediate in the P450 Enzyme CYP121

The P450 enzyme CYP121 from Mycobacterium tuberculosis catalyzes a carbon–carbon (C–C) bond coupling cyclization of the dityrosine substrate containing a diketopiperazine ring, cyclo(l-tyrosine-l-tyrosine) (cYY). An unusual high-spin (S = 5/2) ferric intermediate maximizes its population in less than 5 ms in the rapid freeze-quenching study of CYP121 during the shunt reaction with peracetic acid or hydrogen peroxide in acetic acid solution. Here, we show that this intermediate can also be observed in the crystalline state by EPR spectroscopy. By developing an on-demand-rapid-mixing method for time-resolved serial femtosecond crystallography with X-ray free-electron laser (tr-SFX-XFEL) technology covering the millisecond time domain and without freezing, we structurally monitored the reaction in situ at room temperature. After a 200 ms peracetic acid reaction with the cocrystallized enzyme–substrate microcrystal slurry, a ferric-hydroperoxo intermediate is observed, and its structure is determined at 1.85 Å resolution. The structure shows a hydroperoxyl ligand between the heme and the native substrate, cYY. The oxygen atoms of the hydroperoxo are 2.5 and 3.2 Å from the iron ion. The end-on binding ligand adopts a near-side-on geometry and is weakly associated with the iron ion, causing the unusual high-spin state. This compound 0 intermediate, spectroscopically and structurally observed during the catalytic shunt pathway, reveals a unique binding mode that deviates from the end-on compound 0 intermediates in other heme enzymes. The hydroperoxyl ligand is only 2.9 Å from the bound cYY, suggesting an active oxidant role of the intermediate for direct substrate oxidation in the nonhydroxylation C–C bond coupling chemistry.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗