Structural analysis of spiro beta-lactone proteasome inhibitors

Spiro beta-lactone-based proteasome inhibitors were discovered in the context of an asymmetric catalytic total synthesis of the natural product (+)-lactacystin (1). Lactone 4 was found to be a potent inhibitor of the 26S proteasome, while its C-6 epimer (5) displayed weak activity. Crystallographic studies of the two analogues covalently bound to the 20S proteasome permitted characterization of the important stabilizing interactions between each inhibitor and the proteasome's key catalytic N-terminal threonine residue. This structural data support the hypothesis that the discrepancy in potency between 4 and 5 may be due to differences in the hydrolytic stabilities of the resulting acyl enzyme complexes.     

Engineering Enzymes for Environmental Sustainability

The development and implementation of sustainable catalytic technologies is key to delivering our net-zero targets. Here we review how engineered enzymes, with a focus on those developed using directed evolution, can be deployed to improve the sustainability of numerous processes and help to conserve our environment. Efficient and robust biocatalysts have been engineered to capture carbon dioxide (CO₂) and have been embedded into new efficient metabolic CO₂ fixation pathways. Enzymes have been refined for bioremediation, enhancing their ability to degrade toxic and harmful pollutants. Biocatalytic recycling is gaining momentum, with engineered cutinases and PETases developed for the depolymerization of the abundant plastic, polyethylene terephthalate (PET). Finally, biocatalytic approaches for accessing petroleum-based feedstocks and chemicals are expanding, using optimized enzymes to convert plant biomass into biofuels or other high value products. Through these examples, we hope to illustrate how enzyme engineering and biocatalysis can contribute to the development of cleaner and more efficient chemical industry.     

Biotemplated synthesis of an ordered macroporous superconductor with high critical current density using a cuttlebone template

Periodic superconducting, porous self-supporting monoliths were synthesized using cuttlebone as a morphological template; this produced a lightweight, structurally stable superconductor with a greatly improved critical current density.     

Synthesis and antiplasmodial activity of 3-furyl and 3-thienylquinoxaline-2-carbonitrile 1,4-di-N-oxide derivatives

The aim of this study was to identify new compounds active against Plasmodium falciparum based on our previous research carried out on 3-phenyl-quinoxaline-2-carbonitrile 1,4-di-N-oxide derivatives. Twelve compounds were synthesized and evaluated for antimalarial activity. Eight of them showed an IC(50) less than 1 microM against the 3D7 strain. Derivative 1 demonstrated high potency (IC(50)= 0.63 microM) and good selectivity (SI=10.35), thereby becoming a new lead-compound.     

Separation of inorganic anions on a high capacity porous polymeric monolithic column and application to direct determination of anions in seawater

A commercially available 4.6 mm id x 50 mm polymethacrylate-based monolithic strong anion exchange column (ProSwift SAX-1S) designed for the separation of proteins has been successfully used to separate small inorganic anions in the presence of a seawater sample matrix. Using a hydroxide eluent with suppressed conductivity detection the ion exchange capacity of this column declined over time; however, using KCl as the eluent, the column performance was stable with a capacity of 530 microequiv. for nitrate. The optimum conditions for the separation of iodate, bromate, nitrite, bromide and nitrate were assessed by constructing van Deemter plots using 1.00 and 0.100 M KCl. Efficiencies of up to 26 700 plates/m were recorded using 1.00 M KCl, at a flow rate of 0.20 mL/min but iodate was not baseline resolved from the void peak. By reducing the concentration of the eluent to 0.100 M, efficiencies of up to 39 900 plates/m could be obtained at 0.35 mL/min. By employing a linear gradient ranging from 0.05 to 1.00 M KCl the ions dissolved in distilled water or a salt water matrix could be baseline separated in less than 3 min at a flow rate of 2.50 mL/min.     

An unusual two‐dimensional hydrogen‐bonding network in bis(2,9‐dimethyl‐1,10‐phenanthrolin‐1‐ium) peroxodisulfate dihydrate

The title compound, 2C₁₄H₁₃N₂⁺·S₂O₈²⁻·2H₂O, is a protonated amine salt which is formed from two rather uncommon ionic species, namely a peroxodisulfate (pds²⁻) anion, which lies across a crystallographic inversion centre, and a 2,9‐dimethyl‐1,10‐phenanthrolin‐1‐ium (Hdmph⁺) cation lying in a general position. Each pds²⁻ anion binds to two water molecules through strong water–peroxo O—H...O interactions, giving rise to an unprecedented planar network of hydrogen‐bonded macrocycles which run parallel to (100). The atoms of the large R₈⁸(30) rings are provided by four water molecules bridging in fully extended form (...H—O—H...) and four pds²⁻ anions alternately acting as long (...O—S—O—O—S—O...) and short (...O—S—O...) bridges. The Hdmph⁺ cations, in turn, bind to these units through hydrogen bonds involving their protonated N atoms. In addition, the crystal structure also contains π–π and aromatic–peroxo C—H...O interactions.     

Extremal <Emphasis Type="Italic">G</Emphasis>-invariant eigenvalues of the Laplacian of <Emphasis Type="Italic">G</Emphasis>-invariant metrics

The study of extremal properties of the spectrum often involves restricting the metrics under consideration. Motivated by the work of Abreu and Freitas in the case of the sphere S ² endowed with S ¹-invariant metrics, we consider the subsequence of the spectrum of a Riemannian manifold M which corresponds to metrics and functions invariant under the action of a compact Lie group G. If G has dimension at least 1, we show that the functional λ _k ^G admits no extremal metric under volume-preserving G-invariant deformations. If, moreover, M has dimension at least three, then the functional is unbounded when restricted to any conformal class of G-invariant metrics of fixed volume. As a special case of this, we can consider the standard O(n)-action on S ⁿ ; however, if we also require the metric to be induced by an embedding of S ⁿ in , we get an optimal upper bound on .      

Directed self-assembly in laponite/CdSe/polyaniline nanocomposites

Laponite films provide versatile inorganic scaffolds with materials architectures that direct the self-assembly of CdSe quantum dots (QDs or EviTags) and catalytic surfaces that promote the in situ polymerization of polyaniline (PANI) to yield novel nanocomposites for light emitting diodes (LEDs) and solar cell applications. Water-soluble CdSe EviTags with varying, overlapping emission wavelengths in the visible spectrum were incorporated using soft chemistry routes within Na-Laponite host film platforms to achieve broadband emission in the visible spectrum. QD concentrations, composition and synthesis approach were varied to optimize photophysical properties of the films and to mediate self-assembly, optical cascading and energy transfer. In addition, aniline tetramers coupled to CdSe (QD-AT) surfaces using a dithioate linker were embedded within Cu-Laponite nanoscaffolds and electronically coupled to PANI via vapor phase exposure. Nanotethering and specific host-guest and guest-guest interactions that mediate nanocomposite photophysical behavior were probed using electronic absorption and fluorescence spectroscopies, optical microscopy, AFM, SEM, powder XRD, NMR and ATR-FTIR. Morphology studies indicated that Lap/QD-AT films synthesized using mixed solvent, layer by layer (LbL) methods exhibited anisotropic supramolecular structures with unique mesoscopic ordering that affords bifunctional networks to optimize charge transport.     

The effect of filamentous bacteria on foam production and stability

Bacteria have been implicated in the formation of viscous brown foams that can appear suddenly on wastewater treatment plants. Three strains of the filamentous bacterium Gordonia amarae, isolated from wastewater treatment plants, were investigated to determine their effect on foam formation and stabilisation. During the exponential phase of the bacterial growth a biosurfactant was formed, causing a significant drop in the surface tension of the filtered medium and the formation of persistent foam. Foaming tests in the presence and absence of bacteria showed that bacteria increased foam persistence, most probably by reducing the drainage from the lamellae between bubbles. Experiments showed that > or =55% of the three bacterial strains partitioned into the foam produced by the biosurfactant, indicating that their surfaces were hydrophobic. The extent of partitioning was independent of the growth stage, suggesting that the cell surface hydrophobicity did not change with age, or with cell viability. This work shows that, although the G. amarae cells themselves do not cause foaming, they do produce biosurfactant, which aids foam formation, and they stabilise the foam by reducing the rate of drainage from the foam lamellae.     

Reaction of fluorogenic reagents with proteins I. Mass spectrometric characterization of the reaction with 3-(2-furoyl)quinoline-2-carboxaldehyde, Chromeo P465, and Chromeo P503

3-(2-Furoyl)quinoline-2-carboxaldehyde (FQ), Chromeo P465, and Chromeo P503 are weakly fluorescent reagents that react with primary amines to produce fluorescent products. We studied the reaction of these reagents with alpha-lactalbumin by mass spectrometry. The reaction generated a set of products by the addition of one or more labels to the protein. At room temperature, the reaction was an order of magnitude faster with the Chromeo reagents than with FQ; however, the steady-state labeling efficiency was a factor of two higher for FQ compared with the Chromeo reagents. The relative abundance of the products with FQ usually followed a binomial distribution, which suggests that the labeling sites were uniformly accessible to this reagent. In contrast, the distribution of reaction products with the Chromeo reagents did not follow a binomial distribution for reactions performed in the absence of sodium dodecyl sulfate (SDS); it appears that the protein labeled with the Chromeo reagents refolded into a relatively stable secondary structure that hid some reactive sites. The reaction with the Chromeo reagent did follow the binomial distribution if the protein underwent treatment with 1% SDS at 95 degrees C for 5 min, which apparently disrupts the protein's secondary structure and allowed uniform access to all labeling sites. Chromeo 503 labeled seven of the 13 primary amines in denatured alpha-lactalbumin.