Optical Imaging of Surface Chemistry and Dynamics in Confinement

The interfacial structure and dynamics of water in a microscopically confined geometry is imaged in three dimensions and on millisecond time scales. We developed a 3D wide-field second harmonic microscope that employs structured illumination. We image pH induced chemical changes on the curved and confined inner and outer surfaces of a cylindrical glass micro-capillary immersed in aqueous solution. The image contrast reports on the orientational order of interfacial water, induced by charge-dipole interactions between water molecules and surface charges. The images constitute surface potential maps. Spatially resolved surface pKa,s values are determined for the silica deprotonation reaction. Values range from 2.3<pKa,s<10.7, highlighting the importance of surface heterogeneities. Water molecules that rotate along an oscillating external electric field are also imaged. With this approach, real time movies of surface processes that involve flow, heterogeneities and potentials can be made, which will further developments in electrochemistry, geology, catalysis, biology, and microtechnology

Imaging the Heterogeneity of the Oxygen Evolution Reaction on Gold Electrodes Operando: Activity is Highly Local

Understanding the mechanism of the oxygen evolution reaction (OER), the oxidative half of electrolytic water splitting, has proven challenging. Perhaps the largest hurdle has been gaining experimental insight into the active site of the electrocatalyst used to facilitate this chemistry. Decades of study have clarified that a range of transition-metal oxides have particularly high catalytic activity for the OER. Unfortunately, for virtually all of these materials, metal oxidation and the OER occur at similar potentials. As a result, catalyst surface topography and electronic structure are expected to continuously evolve under reactive conditions. Gaining experimental insight into the OER mechanism on such materials thus requires a tool that allows spatially resolved characterization of the OER activity. In this study, we overcome this formidable experimental challenge using second harmonic microscopy and electrochemical methods to characterize the spatial heterogeneity of OER activity on polycrystalline Au working electrodes. At moderately anodic potentials, we find that the OER activity of the electrode is dominated by <1% of the surface area and that there are two types of active sites. The first is observed at potentials positive of the OER onset and is stable under potential cycling (and thus presumably extends multiple layers into the bulk gold electrode). The second occurs at potentials negative of the OER onset and is removed by potential cycling (suggesting that it involves a structural motif only 1–2 Au layers deep). This type of active site is most easily understood as the catalytically active species (hydrous oxide) in the so-called incipient hydrous oxide/adatom mediator model of electrocatalysis. Combining the ability we demonstrate here to characterize the spatial heterogeneity of OER activity with a systematic program of electrode surface structural modification offers the possibility of creating a generation of OER electrocatalysts with unusually high activity

One-pot bio-synthesis: N-acetyl-d-neuraminic acid production by a powerful engineered whole-cell catalyst

Whole cell biocatalysis is an important tool for pharmaceutical intermediates synthesis, although it is hindered by some shortcomings, such as high cost and toxicity of inducer, mass transfer resistance caused by cell membrane and side reactions. Whole-cell catalysis using N-acetyl-d-glucosamine 2-epimerase (EC 5.1.3.8) and N-acetyl-d-neuraminic acid (Neu5Ac) aldolase (EC 4.1.3.3) is a promising approach for the production of Neu5Ac, a potential precursor of many anti-viral drugs. A powerful catalyst was developed by packaging the enzymes in an engineered bacterium and using a safe temperature-induced vector. Since the mass transfer resistance and the side reactions were substantially reduced, a high Neu5Ac amount (191 mM) was achieved. An efficient method was also presented, which allows one-pot synthesis of Neu5Ac with a safe and economic manner. The results highlight the promise of large-scale Neu5Ac synthesis and point at a potential of our approach as a general strategy to improve whole-cell biocatalysis

Mapping Electrochemical Heterogeneity at Gold Surfaces: A Second Harmonic Imaging Study

Designing efficient catalysts requires correlating surface structure and local chemical composition with reactivity on length scales from nanometers to tens of microns. While much work has been done on this structure/function correlation on single crystals, comparatively little has been done for catalysts of relevance in applications. Such materials are typically highly heterogeneous and thus require methods that allow mapping of the structure/function relationship during electrochemical conversion. Here, we use optical second harmonic imaging combined with cyclic voltammetry to map the surface of gold nanocrystalline and polycrystalline electrodes during electrooxidation and to quantify the spatial extent of surface reconstruction during potential cycling. The wide-field configuration of our microscope allows for real-time imaging of an area ~100 μm in diameter with submicron resolution. By analyzing the voltage dependence of each pixel, we uncover the heterogeneity of the second harmonic signal and quantify the fraction of domains where it follows a positive quadratic dependence with increasing bias. There, the second harmonic intensity is mainly ascribed to electronic polarization contributions at the metal/electrolyte interface. Additionally, we locate areas where the second harmonic signal follows a negative quadratic dependence with increasing bias, which also show the largest changes during successive cyclic voltammetry sweeps as determined by an additional correla-tion coefficient analysis. We assign these areas to domains of higher roughness that are prone to potential-induced surface restructuring, and where anion adsorption occurs at lower potentials than expected based on the CV

Catalytically-active inclusion bodies for biotechnology—general concepts, optimization, and application

Bacterial inclusion bodies (IBs) have long been considered as inactive, unfolded waste material produced by heterologous overexpression of recombinant genes. In industrial applications, they are occasionally used as an alternative in cases where a protein cannot be expressed in soluble form and in high enough amounts. Then, however, refolding approaches are needed to transform inactive IBs into active soluble protein. While anecdotal reports about IBs themselves showing catalytic functionality/activity (CatIB) are found throughout literature, only recently, the use of protein engineering methods has facilitated the on-demand production of CatIBs. CatIB formation is induced usually by fusing short peptide tags or aggregation-inducing protein domains to a target protein. The resulting proteinaceous particles formed by heterologous expression of the respective genes can be regarded as a biologically produced bionanomaterial or, if enzymes are used as target protein, carrier-free enzyme immobilizates. In the present contribution, we review general concepts important for CatIB production, processing, and application