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Hyoscyamine 6β-hydroxylase (H6H) is an iron(II)- and 2-oxoglutarate-dependent (Fe/2OG) oxygenase that produces the prolifically administered antinausea drug, scopolamine. After its namesake hydroxylation reaction, H6H then couples the newly installed C6 oxygen to C7 to produce the drug’s epoxide functionality. Oxoiron(IV) (ferryl) intermediates initiate both reactions by cleaving C–H bonds, but it remains unclear how the enzyme switches the target site and promotes (C6)O–C7 coupling in preference to C7 hydroxylation in the second step. In one possible epoxidation mechanism, the C6 oxygen would─analogously to mechanisms proposed for the Fe/2OG halogenases and, in our more recent study, N-acetylnorloline synthase (LolO)─coordinate as alkoxide to the C7–H-cleaving ferryl intermediate to enable alkoxyl coupling to the ensuing C7 radical. Here, we provide structural and kinetic evidence that H6H does not employ substrate coordination or repositioning for the epoxidation step but instead exploits the distinct spatial dependencies of competitive C–H cleavage (C6 vs C7) and C–O-coupling (oxygen rebound vs cyclization) steps to promote the two-step sequence. Structural comparisons of ferryl-mimicking vanadyl complexes of wild-type H6H and a variant that preferentially 7-hydroxylates instead of epoxidizing 6β-hydroxyhyoscyamine suggest that a modest (∼10°) shift in the Fe–O–H(C7) approach angle is sufficient to change the outcome. The 7-hydroxylation:epoxidation partition ratios of both proteins increase more than 5-fold in 2H2O, reflecting an epoxidation-specific requirement for cleavage of the alcohol O–H bond, which, unlike in the LolO oxacyclization, is not accomplished by iron coordination in advance of C–H cleavage.

Hydrogen fuel cells have drawn increasing attention as one of the most promising next-generation power sources for future automotive transportation. Developing efficient, durable, and low-cost electrocatalysts, to accelerate the sluggish oxygen reduction reaction (ORR) kinetics, is urgently needed to advance fuel cell technologies. Herein, we report on metal–organic frameworks-derived nonprecious dual metal single-atom catalysts (SACs) (Zn/Co–N–C), consisting of Co–N4and Zn–N4local structures. These catalysts exhibited superior ORR activity with a half-wave potential (E1/2) of 0.938 V versus RHE (reversible hydrogen electrode) and robust stability (ΔE1/2= −8.5 mV) after 50k electrochemical cycles. Moreover, this remarkable performance was validated under realistic fuel cell working conditions, achieving a record-high peak power density of ∼1 W cm–2among the reported SACs for alkaline fuel cells. OperandoX-ray absorption spectroscopy was conducted to identify the active sites and reveal catalytic mechanistic insights. The results indicated that the Co atom in the Co–N4structure was the main catalytically active center, where one axial oxygenated species binds to form an Oads–Co–N4moiety during the ORR. In addition, theoretical studies, based on a potential-dependent microkinetic model and core-level shift calculations, showed good agreement with the experimental results and provided insights into the bonding of oxygen species on Co–N4centers during the ORR. This work provides a comprehensive mechanistic understanding of the active sites in the Zn/Co–N–C catalysts and will pave the way for the future design and advancement of high-performance single-site electrocatalysts for fuel cells and other energy applications.

We report the use of iron 1s3p resonant X-ray emission processes to conduct spin-selective, high-energy resolution fluorescence detected X-ray absorption near-edge spectroscopy (HERFD-XANES) on an iron-containing, lithium- and manganese-rich, fully earth-abundant cathode material, Li1.3Mn0.5Fe0.2O2(0.7Li2MnO3·0.3LiFeO2). Coupling this technique with conventional Mn K-edge XANES and detailed extended X-ray absorption fine structure (EXAFS) analysis from both the Mn and Fe vantage points, we gain fundamental insights into the redox processes and migration tendencies of transition metals in this cathode material at the bulk level. We show that during the first charge, Fe3+undergoes oxidation to form Fe4+prior to the activation plateau. Toward the end of activation, a significant fraction of the iron is present as tetrahedral Fe3+. This observation reveals that iron migration from octahedral to tetrahedral sites and iron reduction are initiated during activation. Upon first discharge from the activated state, a continuous and overlapping reduction of both Fe and Mn is observed, with Fe largely restored back as an octahedrally coordinated Fe3+. The manganese local environment gradually changes to a distorted cooperative Jahn–Teller Mn3+structure during discharge, with the clear presence of two Mn–O as well as two Mn–Mn correlation distances at 2.0 V. The significant reduction of manganese in the very first discharge is distinctly different from that seen in typical nickel-based lithium-manganese-rich materials but is similar to that observed for pure Li2MnO3. These findings shed light on key structure–property correlations in the cathode material and point to a causative relationship between the redox mechanisms as well as structural changes endured by the material and relatively poor performance during extended electrochemical cycling.

Transition metal nitride (TMN-) based materials have recently emerged as promising non-precious-metal-containing electrocatalysts for the oxygen reduction reaction (ORR) in alkaline media. However, the lack of fundamental understanding of the oxide surface has limited insights into structure–(re)activity relationships and rational catalyst design. Here we demonstrate how a well-defined TMN can dictate/control the as-formed oxide surface and the resulting ORR electrocatalytic activity. Structural characterization of MnN nanocuboids revealed that an electrocatalytically active Mn3O4shell grew epitaxially on the MnN core, with an expansive strain along the [010] direction to the surface Mn3O4. The strained Mn3O4shell on the MnN core exhibited an intrinsic activity that was over 300% higher than that of pure Mn3O4. A combined electrochemical and computational investigation indicated/suggested that the enhancement probably originates from a more hydroxylated oxide surface resulting from the expansive strain. This work establishes a clear and definitive atomistic picture of the nitride/oxide interface and provides a comprehensive mechanistic understanding of the structure–reactivity relationship in TMNs, critical for other catalytic interfaces for different electrochemical processes.

Advances in electrocatalysis research rely heavily on building a thorough mechanistic understanding of catalyst active sites under realistic operating conditions. Only recently have techniques emerged that enable sensitive spectroscopic data collection to distinguish catalytically relevant surface sites from the underlying bulk material under applied potential in the presence of an electrolyte layer. Here, we demonstrate that operandohigh-energy-resolution fluorescence detected X-ray absorption spectroscopy (HERFD-XAS) is a powerful spectroscopic method which offers critical surface chemistry insights in CO2electroreduction with sub-electronvolt energy resolution using hard X-rays. Combined with the high surface area-to-volume ratio of 5 nm copper nanoparticles, operandoHERFD-XAS allows us to observe with clear evidence the breaking of chemical bonds between the ligands and the Cu surface as part of the ligand desorption process occurring under electrochemical potentials relevant for the CO2reduction reaction (CO2RR). In addition, the dynamic evolution of oxidation state and coordination number throughout the operation of the nanocatalyst was continuously tracked. With these results in hand, undercoordinated metallic copper nanograins are proposed to be the real active sites in the CO2RR. This work emphasizes the importance of HERFD-XAS compared to routine XAS in catalyst characterization and mechanism exploration, especially in the complicated electrochemical CO2RR.

Valence-to-core X-ray emission spectroscopy (VtC XES) is an emerging technique that uses hard X-rays to probe the valence electronic structure of an absorbing atom. Despite finding varied applications for light elements and first row transition metals, little work has been done on heavier elements such as second and third row transition metals. This lack of application is at least partially due to the relatively low resolution of the data at the high energies required to measure these elements, which obscures the useful chemical information that can be extracted from the lower energy, higher resolution spectra of lighter elements. Herein, we collect data on a set of platinum-containing compounds and demonstrate that the VtC XES resolution can be dramatically enhanced by exciting the platinum atom in resonance with its L3-edge white line absorption. Whereas spectra excited using standard nonresonant absorption well above the Pt L3-edge display broad, unfeatured VtC regions, resonant XES (RXES) spectra have more than twofold improved resolution and are revealed to be rich in chemical information with the ability to distinguish between even closely related species. We further demonstrate that these RXES spectra may be used to selectively probe individual components of a mixture of Pt-containing compounds, establishing this technique as a viable probe for chemically complex samples. Lastly, it is shown that the spectra are interpretable using a molecular orbital framework and may be calculated using density functional theory, thus suggesting resonant excitation as a general strategy for extracting chemically useful information from heavy element VtC spectra.

Carbon dioxide electroreduction facilitates the sustainable synthesis of fuels and chemicals1. Although Cu enables CO2-to-multicarbon product (C2+) conversion, the nature of the active sites under operating conditions remains elusive2. Importantly, identifying active sites of high-performance Cu nanocatalysts necessitates nanoscale, time-resolved operando techniques3–5. Here, we present a comprehensive investigation of the structural dynamics during the life cycle of Cu nanocatalysts. A 7 nm Cu nanoparticle ensemble evolves into metallic Cu nanograins during electrolysis before complete oxidation to single-crystal Cu2O nanocubes following post-electrolysis air exposure. Operando analytical and four-dimensional electrochemical liquid-cell scanning transmission electron microscopy shows the presence of metallic Cu nanograins under CO2reduction conditions. Correlated high-energy-resolution time-resolved X-ray spectroscopy suggests that metallic Cu, rich in nanograin boundaries, supports undercoordinated active sites for C–C coupling. Quantitative structure–activity correlation shows that a higher fraction of metallic Cu nanograins leads to higher C2+selectivity. A 7 nm Cu nanoparticle ensemble, with a unity fraction of active Cu nanograins, exhibits sixfold higher C2+selectivity than the 18 nm counterpart with one-third of active Cu nanograins. The correlation of multimodal operando techniques serves as a powerful platform to advance our fundamental understanding of the complex structural evolution of nanocatalysts under electrochemical conditions.

The iron(II)- and 2-(oxo)glutarate-dependent (Fe/2OG) oxygenases catalyze an array of challenging transformations via a common iron(IV)-oxo (ferryl) intermediate, which in most cases abstracts hydrogen (H•) from an aliphatic carbon of the substrate. Although it has been shown that the relative disposition of the Fe–O and C–H bonds can control the rate of H• abstraction and fate of the resultant substrate radical, there remains a paucity of structural information on the actual ferryl states, owing to their high reactivity. We demonstrate here that the stable vanadyl ion [(VIV-oxo)2+] binds along with 2OG or its decarboxylation product, succinate, in the active site of two different Fe/2OG enzymes to faithfully mimic their transient ferryl states. Both ferryl and vanadyl complexes of the Fe/2OG halogenase, SyrB2, remain stably bound to its carrier protein substrate (l-aminoacyl-S-SyrB1), whereas the corresponding complexes harboring transition metals (Fe, Mn) in lower oxidation states dissociate. In the well-studied taurine:2OG dioxygenase (TauD), the disposition of the substrate C–H bond relative to the vanadyl ion defined by pulse electron paramagnetic resonance methods is consistent with the crystal structure of the reactant complex and computational models of the ferryl state. Vanadyl substitution may thus afford access to structural details of the key ferryl intermediates in this important enzyme class.

Iron(II)- and 2-oxoglutarate-dependent (Fe/2OG) oxygenases generate iron(IV)-oxo (ferryl) intermediates that can abstract hydrogen from aliphatic carbons (R–H). Hydroxylation proceeds by coupling of the resultant substrate radical (R•) and oxygen of the Fe(III)-OH complex (“oxygen rebound”). Nonhydroxylation outcomes result from different fates of the Fe(III)-OH/R• state; for example, halogenation results from R• coupling to a halogen ligand cis to the hydroxide. We previously suggested that halogenases control substrate-cofactor disposition to disfavor oxygen rebound and permit halogen coupling to prevail. Here, we explored the general implication that, when a ferryl intermediate can ambiguously target two substrate carbons for different outcomes, rebound to the site capable of the alternative outcome should be slower than to the adjacent, solely hydroxylated site. We evaluated this prediction for (i) the halogenase SyrB2, which exclusively hydroxylates C5 of norvaline appended to its carrier protein but can either chlorinate or hydroxylate C4 and (ii) two bifunctional enzymes that normally hydroxylate one carbon before coupling that oxygen to a second carbon (producing an oxacycle) but can, upon encountering deuterium at the first site, hydroxylate the second site instead. In all three cases, substrate hydroxylation incorporates a greater fraction of solvent-derived oxygen at the site that can also undergo the alternative outcome than at the other site, most likely reflecting an increased exchange of the initially O2-derived oxygen ligand in the longer-lived Fe(III)-OH/R• states. Suppression of rebound may thus be generally important for nonhydroxylation outcomes by these enzymes.

Understanding the mechanisms of charge transport in ternary and higher order spinels would enable design principles for enhancing their electronic and electrochemical properties for energy applications. Here we investigate the ternary CoxMn3–xO4spinel system and determine the relationship between electronic properties and cation site occupation, based on the octahedral site, donor–acceptor formalization introduced in the polaron hopping model. We employ synchrotron X-ray emission spectroscopy to characterize the donor–acceptor cation pairs in the lattice. We find that the stochiometric volcano trend in electronic conductivity correlates well with the concentration of Mn4+/Mn3+hopping pairs, while the concentration of Mn3+/Mn2+pairs is unchanged with stoichiometry. We also find that Co does not directly contribute to conductivity; however, the Co does create configurational disorder that leads to the generation of hopping pairs for Mn at octahedral sites. From these results we conclude that Mn4+/Mn3+pairs are the dominant active species and the origin of the stoichiometry-dependent behavior for polaron charge transport. This work provides a starting point for understanding and optimizing charge transport for higher order spinels.

A ribonucleotide reductase (RNR) from Flavobacterium johnsoniae(Fj) differs fundamentally from known (subclass a–c) class I RNRs, warranting its assignment to a new subclass, Id. Its β subunit shares with Ib counterparts the requirements for manganese(II) and superoxide (O2–) for activation, but it does not require the O2–-supplying flavoprotein (NrdI) needed in Ib systems, instead scavenging the oxidant from solution. Although Fjβ has tyrosine at the appropriate sequence position (Tyr 104), this residue is not oxidized to a radical upon activation, as occurs in the Ia/b proteins. Rather, Fjβ directly deploys an oxidized dimanganese cofactor for radical initiation. In treatment with one-electron reductants, the cofactor can undergo cooperative three-electron reduction to the II/II state, in contrast to the quantitative univalent reduction to inactive “met” (III/III) forms seen with I(a–c) βs. This tendency makes Fjβ unusually robust, as the II/II form can readily be reactivated. The structure of the protein rationalizes its distinctive traits. A distortion in a core helix of the ferritin-like architecture renders the active site unusually open, introduces a cavity near the cofactor, and positions a subclass-d-specific Lys residue to shepherd O2–to the Mn2II/IIcluster. Relative to the positions of the radical tyrosines in the Ia/b proteins, the unreactive Tyr 104 of Fjβ is held away from the cofactor by a hydrogen bond with a subclass-d-specific Thr residue. Structural comparisons, considered with its uniquely simple mode of activation, suggest that the Id protein might most closely resemble the primordial RNR-β.

X-ray crystallography at X-ray free-electron laser sources is a powerful method for studying macromolecules at biologically relevant temperatures. Moreover, when combined with complementary techniques like X-ray emission spectroscopy, both global structures and chemical properties of metalloenzymes can be obtained concurrently, providing insights into the interplay between the protein structure and dynamics and the chemistry at an active site. The implementation of such a multimodal approach can be compromised by conflicting requirements to optimize each individual method. In particular, the method used for sample delivery greatly affects the data quality. We present here a robust way of delivering controlled sample amounts on demand using acoustic droplet ejection coupled with a conveyor belt drive that is optimized for crystallography and spectroscopy measurements of photochemical and chemical reactions over a wide range of time scales. Studies with photosystem II, the phytochrome photoreceptor, and ribonucleotide reductase R2 illustrate the power and versatility of this method.

This review considers the likely advances in plant and animal biotechnology over the short and medium term. It attempts to set these in context with technology-driven advances in management. Consideration is given to the extent to which current political and social considerations will mitigate against the widespread adoption of these advances.

This paper originates from a presentation at the International Conference on Assimilate Transport and Partitioning, Newcastle, NSW, August 1999 In order to investigate the roles of different cell types, metabolite compartmentation in barley (Hordeum vulgare L.) leaf tissue was mapped at the single-cell level, using single-cell sampling and analysis (SiCSA) techniques. The partitioning of recently fixed photoassimilate was investigated for the first time at single-cell resolution, using BAMS (biological accelerator mass spectroscopy) for precise measurement of 14C in femtomole quantities. The data obtained by BAMS qualitatively reflect concentrations of sugars in different cell types measured by SiCSA. Calculation of 14C-specific activities showed that the radioactive label saturated the mesophyll and parenchymatous bundle sheath (PBS) pools within the 45-min labelling period. During the photoperiod, sucrose concentration increased to 200 mM in mesophyll cells. The concentration of malate also increased during the photoperiod in mesophyll and PBS cells. Epidermal cells contained very low concentrations of sugar but high concentrations of malate (120–180 mM) and did not show significant diurnal changes. Accumulation of sugars and fructan synthesis could be induced in mesophyll and PBS cells by reduced export of sugars from leaves or, alternatively, when sugars were supplied from excised leaf blade bases immersed in a sucrose solution in the dark. The epidermis accumulated additional malate in step with the accumulation of sugar by the mesophyll/PBS cells during the long-term reduction of export. Immunolocalisation of Rubisco and cytochrome oxidase proteins was used to analyse the distribution of enzymes of photoassimilation and respiration between functionally different cells in mature leaves of barley.

The article discusses technological developments in workplace design, particularly in the U.S., as of mid-February 2014. Based on the Post-Occupancy Evaluation (POE) Survey research by the Center for the Built Environment, each staff is subjective in his/her own experiences and reactions to a busy environment. The study of top firms by design firms like Gensler showed that employees need spaces to accommodate four work modes, including socialization, learning, and focus.

The article focuses on efforts to achieve acoustical privacy by decreasing noise pollution in open plan offices, and refers to impact of noise distraction on productivity, need for collaboration and separation in office layout, and list of suggestions which includes masking of sound, filling gaps in glass partitions, and educating the staff.

Soluble acid invertase from wheat coleoptiles was purified to electrophoretic homogeneity. A comparison of molecular weight by SDS-PAGE and gel filtration suggested that the enzyme was a monomer of Mr50 000. The enzyme was a glycoprotein and, after chemical deglycosylation, possessed a Mrof 48000. A polyclonal antiserum was raised against the deglycosylated protein. This cross-reacted specifically with acid invertase. A putative precursor of invertase synthesized in a cell-free translation system was detected by SDS-PAGE and fluorography of the immunoprecipitated polypeptides. The distribution of acid invertase in wheat seedling shoots was investigated both by visualizing invertase activity after starch gel electrophoresis and by immunoblotting. Both techniques identified two forms of invertase in extracts of the primary leaf and only one form in extracts of coleoptiles. The low pH optimum and the glycoprotein nature of wheat coleoptile invertase are consistent with a vacuolar location. Fructose inhibited its activity, suggesting that enzyme activity could be modulated by end-product inhibition.