Advances In Structural Characterization Research Articles

Molecular insights of nanozymes from design to catalytic mechanism.

Emerging as cost-effective potential alternatives to natural enzymes, nanozymes have attracted increasing interest in broad fields. To exploit the in-depth potential of nanozymes, rational structural engineering and explicit catalytic mechanisms at the molecular scale are required. Recently, impressive progress has been made in mimicking the characteristics of natural enzymes by constructing metal active sites, binding pockets, scaffolds, and delicate allosteric regulation. Ingenious in-depth studies have been conducted with advances in structural characterization and theoretical calculations, unveiling the "black box" of nanozyme-catalytic mechanisms. This review introduces the state-of-art synthesis strategies by learning from the natural enzyme counterparts and summarizes the general overview of the nanozyme mechanism with a particular emphasis on the adsorbed intermediates and descriptors that predict the nanozyme activity The emerging activity assessment methodology that illustrates the relationship between electrochemical oxygen reduction and enzymatic oxygen reduction is discussed with up-to-date advances Future opportunities and challenges are presented in the end to spark more profound work and attract more researchers from various backgrounds to the flourishing field of nanozymes.

Recent advances in structural characterization of biomacromolecules in foods via small-angle X-ray scattering.

Small-angle X-ray scattering (SAXS) is a method for examining the solution structure, oligomeric state, conformational changes, and flexibility of biomacromolecules at a scale ranging from a few Angstroms to hundreds of nanometers. Wide time scales ranging from real time (milliseconds) to minutes can be also covered by SAXS. With many advantages, SAXS has been extensively used, it is widely used in the structural characterization of biomacromolecules in food science and technology. However, the application of SAXS in charactering the structure of food biomacromolecules has not been reviewed so far. In the current review, the principle, theoretical calculations and modeling programs are summarized, technical advances in the experimental setups and corresponding applications of in situ capabilities: combination of chromatography, time-resolved, temperature, pressure, flow-through are elaborated. Recent applications of SAXS for monitoring structural properties of biomacromolecules in food including protein, carbohydrate and lipid are also highlighted, and limitations and prospects for developing SAXS based on facility upgraded and artificial intelligence to study the structural properties of biomacromolecules are finally discussed. Future research should focus on extending machine time, simplifying SAXS data treatment, optimizing modeling methods in order to achieve an integrated structural biology based on SAXS as a practical tool for investigating the structure-function relationship of biomacromolecules in food industry.

PEMFC Catalyst Layers: Recent Advances in Process-Structure-Property Correlations

Proton exchange membrane (PEM) fuel cells are being developed as alternative energy sources for both residential and automotive applications. In order for this technology to become fully commercial, reduction of cost and improvements in performance and durability of PEM fuel cells’ membrane electrode assemblies (MEAs) are still required. To address the requirements for further cost reduction, structure and composition of the cathode catalyst layer (CCL) has to be optimized in order to achieve optimum performance and durability while minimizing the platinum group metals (PGM) loading. In order to rationally design CCL structures that meet the performance and durability requirements, a better understanding of structure versus performance relationships is needed. This requires the capability to fabricate different CCL structures, to characterize the spatial distribution of all the phases within the catalyst layer1-3 (carbon, Pt, ionomer, and void), to measure the physico-chemical properties (both ex-situ and in-situ), and, finally, to use these experimental data as inputs for the development of a model-based understanding of the relationship between the CCL structure and CCL performance and durability. Recently, significant progress has been made in reliable measurement and modeling of key CCL properties such as thermal conductivity, electronic conductivity, and gas diffusivity4-7. Dependence of these properties on the CCL structure and manufacturing processes provides support for rational CCL design. References J. Wu, A.P. Hitchcock, M. Lerotic, D. Shapiro, V. Berejnov, D. Susac, J. Stumper, “4D imaging of polymer electrolyte membrane fuel cell cathodes by scanning X-ray microscopy”, Microsc. Microanal., 23, 1784 (2017).V. Berejnov, M. Saha, D. Susac, J. Stumper, M. West, A.P. Hitchcock, Advances in structural characterization using soft x-ray scanning transmission microscopy (STXM): Mapping and measuring porosity in PEM-FC catalyst layers, ECS Transactions, 80, 241 (2017).L.G.A. Melo, A.P. Hitchcock, J. Jankovic, J. Stumper, D. Susac, V. Berejnov, Quantitative mapping of ionomer in catalyst layers by electron and x-ray spectromicroscopy, ECS Transactions, 80, 275 (2017).M. Ahadi, M. Tam, M.S. Saha, J. Stumper, M. Bahrami, Thermal conductivity of catalyst layer of polymer electrolyte membrane fuel cells: Part 1 – Experimental study, J. Power Sources, 354, 207 (2017).M. Ahadi, A. Putz, J. Stumper, M. Bahrami, Thermal conductivity of catalyst layer of polymer electrolyte membrane fuel cells: Part 2 – Analytical modeling, J. Power Sources, 354, 215 (2017).M. Ahadi, M. Tam, A. Putz, J. Stumper, C. McCague, M. Bahrami, Electrical conductivity of PEM fuel cell catalyst layers: Through-plane vs. in-plane, in: European Hydrogen Energy Conference 2018, Costa del Sol, Spain, 14-16th March, paper no. (124), 206 (2018).S. Salari, C. McCague, M. Tam, M.S. Saha, J. Stumper, M. Bahrami, Accurate ex-situ measurements of PEM fuel cells catalyst layer dry diffusivity, ECS Transactions, 69, 419 (2015).

Probing Structural Variation and Dynamics in the HIV-1 Env Fusion Glycoprotein.

Recent advances in structural characterization of the HIV envelope glycoprotein (Env) have provided a high-resolution glimpse of the architecture of this target for neutralizing antibodies and the machinery responsible for mediating receptor binding and membrane fusion. These structures primarily capture the detailed organization of the receptor-naive, prefusion conformation of Env, but under native solution conditions Env is highly dynamic, sampling multiple conformational states as well as exhibiting local protein flexibility. Special emphasis is placed on the use of biophysical methods, including single-molecule fluorescence microscopy and hydrogen/deuterium-exchange mass spectrometry. Using novel biophysical approaches, striking isolate-specific differences in Env's dynamic profile have been revealed that appear to underlie phenotypic differences of the viral isolates such as neutralization sensitivity and CD4 receptor reactivity. Structural studies are complemented by novel biophysical investigations that enable visualization of the dynamics of HIV-1 Env under native conditions. These approaches will also enable us to gain new insights into the mechanisms of action of antibodies and drugs.

Advances in Structural Characterization Using Soft X-ray Scanning Transmission Microscopy (STXM): Mapping and Measuring Porosity in PEM-FC Catalyst Layers

Access of fuel and oxidant gases to the active catalyst sites in dispersive electrodes of polymer electrolyte membrane fuel cells (PEM-FC) is an important characteristic defining the performance of the whole catalyst coated membrane unit (CCM). The electrode porosity – the ratio of empty space to the whole volume of the electrode – is a metric widely used to characterize the ability of a particular electrode to provide gas transport. It is common practice to present electrode porosity as a number averaged over a macroscopic volume of the electrode [1]. In compliment to already established methods, soft X-ray scanning transmission X-ray microscopy (STXM) offers the possibility to visualize the microscopic network of the pore space in the electrode with ~30 nm spatial resolution. Thus, in addition to average porosity, STXM provides porosity distributions in electrodes, with a local value averaged per voxel of ~1003 nm3. Relative to studies of the electrode porosity visualized by the focused ion beam – scanning electron microscopy method (FIB-SEM) [2], STXM provides direct visualization of the pore-space in the electrode without destroying the sample and omitting the 3D reconstruction process. Here we present STXM porosity distribution for 2D projection of the thin cathode lamella. STXM pore space visualization relies on filling the pores with a material that STXM can discriminate spectroscopically relative to the other components (carbon support, ionomer, and metal catalyst-Pt) [3]. In these experiments the cathode catalyst decals are embedded with a custom epoxy (TTE, the product of reacting trimethylolpropane triglycidyl ether and 4,4’-methylenebis(2-methylcyclohexylamine in 1:1 wt ratio, which results in a final chemical formula (C10H10N3)n and density of 1.0 g/cm3). While the TTE epoxy is fluid it wets well the electrode inner interfaces filling most of the available space. 100 nm thick microtomed sections of the cathode embedded in TTE are measured by STXM at the C 1s edge at the Canadian Light Source (Saskatoon) or the Advanced Light Source (Berkeley). The C1s stacks, consisting of images at 50-70 energies over 280-330 eV with 100 nm pixels, are converted to optical density, and then fit using standard, quantitative reference spectra (OD1 scale) of carbon-support (Cs), ionomer (I), TTE and Pt components to construct 2D component maps of each component. The grayscale value at each pixel is the nm thickness of that component. The total volume is the sum of Cs, I, TTE, and Pt values at each pixel. The 2D porosity map is then the ratio of the TTE map to the total-volume map, where each pixel has a unit of part per volume (ppvol). STXM performed using BL 10ID1 at CLS and BL 5.3.2.2 at ALS. Research supported by NSERC. S. Shukla, et.al., 231 ECS 2017, Abst 97034, https://ecs.confex.com/ecs/231/webprogram/Paper97034.htmlM.Sabharwal, et al., Fuel Cell, 16 (2016) No.6, 734–753H. Ade, et al., Polymers, 49 (2008) 643-675 Figure 1

Advances in Structural Characterization Using Soft X-ray Scanning Transmission Microscopy (STXM): Mapping and Measuring Porosity in PEM-FC Catalyst Layers

A new method for characterizing porosity in polymer electrolyte membrane fuel cell (PEM-FC) catalyst layers (CL) is presented. The CL porosity is derived from the distribution of TTE, a non-swelling embedding epoxy, which is discriminated by STXM. Analysis of carbon 1s image sequences of TTE-embedded PEM FC microtome sections measured by STXM generates thickness maps of the CL major components: carbon support, ionomer, TTE, and Pt. The sum of all maps yields a 2D model of the CL total thickness. The TTE-porosity map is the ratio of the TTE thickness map to the total thickness map. We show that TTE-porosity maps are sensitive to CL processing. Examples illustrating how TTE-porosity can help optimize CL processing are presented.

Invited: Advances in Structural Characterization of PEM Fuel Cell Catalyst Layers By Soft X-Ray Scanning Transmission Microscopy

Proton exchange membrane fuel cells (PEMFCs) are among the most attractive alternative energy sources for automotive application due to their high power density, high efficiency, and potential to significantly reduce adverse environmental impact. However, high material costs and durability issues remain major challenges for large scale commercialization [1, 2]. The catalyst coated membrane (CCM), as a main component in PEMFCs, that employs expensive Pt based porous electrode layers responsible for the generation of pathways for reactant transport enabling efficient electron and proton conductivity and managing product water removal. Improved understanding of the relationship between material properties, electrode structure and overall fuel cell performance is essential for material and process down-selection in order to establish the most cost effective PEMFC manufacturing process. For that reason, the development of new, non destructive characterization methods to allow evaluation of catalyst layer composition and microstructure with high spatial resolution and individual component mapping is needed. In recent years, a novel synchrotron based technique: soft X-ray scanning transmission microscopy (STXM) has emerged and extended the number of standard available microscopic characterization tools and significantly improving our understanding of how materials are distributed from micro to nano-scales in fuel cell electrodes. STXM is able to image a fuel cell catalyst layer and differentiate each chemically specific material component due to its near edge X-ray specific absorption (NEXAFS) signature [3-11]. The spatial resolution of this method is 30 nm. We have developed several STXM applications to characterize a) materials in the dry CCM in 2D cross sections, b) in 3D, and c) while applying environmental changes in 2D. Using two-energy mapping at F 1s and C 1s edges, a 2D material reconstruction methodology for catalyst layer micro-structural characterization and ionomer mapping was developed [7]. Both conventional and nanostructured thin film (NSTF) based catalyst layers are being investigated [9, 10]. Applying a tomographic approach, a STXM spectro-tomography method was established and proven to provide a 3D reconstruction of the cathode catalyst layer with spatially resolved carbon and ionomer species [8]. Due to its ability to spectroscopically differentiate gas, liquid and solid water using the O 1s absorption edge, STXM is being applied to study hydrated CCMs [7]. An environmental wet cell for in situSTXM studies of CCM components under controllable temperature and relative humidity conditions has been developed and used to map all three phases of water in the electrode under various conditions including those relevant to cold start. In this paper and the coming talk, we will review progress in development of STXM methodology for characterization of catalyst layer structures. Efforts in deriving parameters to aid structure-properties-performance correlations will be discussed.STXM measurements are carried out at the Canadian Light Source and at the Advanced Light source.

Two ATPases

In this article, I reflect on research on two ATPases. The first is F(1)F(0)-ATPase, also known as ATP synthase. It is the terminal enzyme in oxidative phosphorylation and famous as a nanomotor. Early work on mitochondrial enzyme involved purification in large amount, followed by deduction of subunit composition and stoichiometry and determination of molecular sizes of holoenzyme and individual subunits. Later work on Escherichia coli enzyme utilized mutagenesis and optical probes to reveal the molecular mechanism of ATP hydrolysis and detailed facets of catalysis. The second ATPase is P-glycoprotein, which confers multidrug resistance, notably to anticancer drugs, in mammalian cells. Purification of the protein in large quantity allowed detailed characterization of catalysis, formulation of an alternating sites mechanism, and recently, advances in structural characterization.

Recent Advances in Structural Characterization of Natural Rubber

Abstract Structural characterization of rubber from Hevea brasiliensis (NR) has been carried out to elucidate the biosynthesis mechanism of rubber molecule as well as to find the relationship between physical properties characteristic of NR and its structure. Recent advances of structural studies have provided a series of new information on the structure of long-chain branching based on the result of selective decomposition of branch-points by chemical and enzymatic treatments as well as physical treatment such as polar solvent treatments and washing of NR latex by centrifugation in the presence of a surfactant. The measurement of the resulting rubber with NMR, FTIR, SEC and dilute solution viscosity provided confirming evidence that the initiating terminal (ω-terminal) with an unidentified functional group and phosphate terminal (α-terminal) form branch-points by hydrogen bond, ionic bond or micelle formation of phospholipids linked to both terminal groups. Based on these results, the origin of green properties characteristic of NR has been explained and a new mechanism of storage hardening has been proposed. The purification method by saponification of NR latex developed for the structural characterization has been applied to produce purified NR latex free from Type I allergic reaction. Instantaneous coagulation of saponified latex by the use of a flocculant and formic acid has provided solid saponified NR having good green and cured rubber properties.

Advances in Structural Characterization of Thin Film Nanocrystalline Silicon for Photovoltaic Applications

The knowledge and control of the structural and morphological properties of nanocrystalline silicon is a fundamental requisite for its proper application in photovoltaics. To this purpose, nanocrystalline silicon films grown by Low Energy Plasma Enhanced Chemical Vapour Deposition (LEPECVD) technique on different kinds of substrates were submitted to a systematic characterization using Raman spectroscopy, X-ray diffraction (XRD) and high-resolution transmission electron microscopy (HRTEM). The results showed that the nature of the film substrate induces deep changes in the structural properties of the deposited films. The importance of a Raman in–depth analysis for an accurate determination of the sample structure has been also demonstrated.

Phase contrast imaging and diffraction-space mapping of periodic inversion domains in LiNbO3 with advanced x-rays

This paper presents some recent advances in structural characterization of periodic inversion domains in ferroelectric nonlinear optical crystals using novel techniques such as diffraction-space mapping, diffuse scattering imaging and phase-contrast imaging. The great advantages of using both highly coherent hard x-ray photons delivered by the third-generation synchrotron source and the well-defined x-ray beam provided by state-of-the-art laboratory diffractometer are addressed in terms of investigation of periodically domain-inverted LiNbO3 crystals. The microstructural characteristics of the periodic inversion domains in LiNbO3 are discussed, with an indication of their differences from naturally-occurring inversion domains.

Nucleation and crystallization of globular proteins — what we know and what is missing

Recently, much progress has been made in understanding the nucleation and crystallization of globular proteins, including the formation of compositional and structural crystal defects. Insight into the interactions of (screened) protein macro-ions in solution, obtained from light scattering, small angle X-ray scattering and osmotic pressure studies, can guide the search for crystallization conditions. These studies show that the nucleation of globular proteins is governed by the same principles as that of small molecules. However, failure to account for direct and indirect (hydrodynamic) protein interactions in the solutions results in unrealistic aggregation scenarios. Microscopic studies of numerous proteins reveal that crystals grow by the attachment of growth units through the same layer-spreading mechanisms as inorganic crystals. Investigations of the growth kinetics of hen-egg-white lysozyme (HEWL) reveal non-steady behavior under steady external conditions. Long-term variations in growth rates are due to changes in step-originating dislocation groups. Fluctuations on a shorter timescale reflect the non-linear dynamics of layer growth that results from the interplay between interfacial kinetics and bulk transport. Systematic gel electrophoretic analyses suggest that most HEWL crystallization studies have been performed with material containing other proteins at percent levels. Yet, sub-percent levels of protein impurities impede growth step propagation and play a role in the formation of structural/compositional inhomogeneities. In crystal growth from highly purified HEWL solutions, however, such inhomogeneities are much weaker and form only in response to unusually large changes in growth conditions. Equally important for connecting growth conditions to crystal perfection and diffraction resolution are recent advances in structural characterization through high-resolution Bragg reflection profiling and X-ray topography.

Recent Advances in Structural Characterization of Elastomers

Abstract One of the main targets of the structural characterization of elastomers so far has been the correlation of the polymerization conditions with the properties of the resulting polymers. The first step is the analysis of polymer structure, such as the chemical composition of copolymers, isomeric structure of diene polymers, degree of branching, extent of modification, functionality of end groups, amounts of abnormal groups, tacticity, and so on. Progress in nuclear magnetic resonance spectroscopy (NMR) makes possible the second step, which is the structural characterization of polymer chains, such as the sequence distribution of comonomer units in copolymer, isomeric units in diene polymers, configurational sequences in vinyl polymers, head and tail arrangement of monomer units. Recent development of FT-NMR spectroscopy, high-field spectroscopy from 300 MHz to 600 MHz at 1H-NMR, solid-state 13C-NMR, and two-dimensional NMR has facilitated a more detailed analysis of these structural features. The complexity of the structure of elastomers, which is derived from highly controlled copolymerization processes, leads to the widespread application of modern FT-NMR spectroscopy. It may reasonably be said that a fair number of structural problems in elastomers has been solved by NMR analysis. The high sensitivity of Fourier-transform infrared spectroscopy (FT-IR) has enabled one to determine trace structural changes in elastomers. Coupled on-line techniques such as gas-chromatography-mass-spectrometry combined with pyrolysis, liquid chromatography-NMR, and gel permeation chromatography-FT-IR will be powerful tools for the characterization of elastomers. Progress in analytical instruments has stimulated the development of high-performance elastomers, the synthesis of new speciality elastomers, and the elucidation of mechanisms for property enhancements. The use of modern instruments and a combination of ordinary methods of structural analysis have satisfied needs to some extent. However, a newer method of structural characterization is always desired in order to achieve higher orders of information. for example, the characterization of inhomogeneity along the polymer chain and between the polymer chains has become an important problem, especially in polymer blend systems. As to the former problem, spectroscopic methods provide only limited information. Although the NMR and FT-IR spectroscopies are very powerful tools for the analysis of short sequence distributions, it is difficult to characterize the distribution of long sequences and hybrid systems containing random and blocked sequences along the polymer chain. Gel permeation chromatography (GPC) is one of the most popular techniques for the analysis of molecular-weight distribution. However, it provides complicated information including molecular-weight distribution and chemical-composition distribution in the case of copolymers. Recent progress of high-performance liquid chromatography (HPLC) has provided a new powerful tool for the structural characterization of copolymers. It is appropriate to review the recent advances in structural characterization of elastomers, especially the characterization of microstructure of polymer chains, from the viewpoints of methodology and applicability of new methods. As to the NMR analysis of elastomers, reviews are available. Here, considerable attention is focused on the procedures for the assignment of signals, because the applicability of a NMR method is based on the reliability of the signal assignments. The other topics are selected to provide direct information for polymer synthesis or polymer properties.