Assessing the Binding Performance of Amyloid–Carbon Membranes toward Heavy Metal Ions

Amyloid–carbon hybrid membranes have exceptional performance in removing heavy metal ions from water because of the presence of multiple binding sites on the amyloid fibrils, but the binding process is still not fully understood. To understand the mechanisms of amyloid–metal ion binding, we perform adsorption isotherms on a model system given by β-lactoglobulin amyloid fibrils and four representative heavy metal ions: chromium (Cr), nickel (Ni), silver (Ag), and platinum (Pt). Furthermore, to get a comprehensive thermodynamic picture of the binding process between amino acid residues and heavy metals, we here use isothermal titration calorimetry on native β-lactoglobulin monomers and amyloid fibrils exposed to the two model metal ions, that is, silver and chromium. A conclusive thermodynamic insight on the binding process emerges by direct measurements of enthalpy and entropy changes, association binding constant, and average number of binding sites of the protein monomer and amyloid fibril. As a result of the strong amyloid binding affinity between amino acids and metal ions, when the protein is converted into amyloid fibrils and assembled into membranes, the resulting amyloid-activated carbon hybrids remove all the tested heavy metals with efficiencies beyond 99%. Importantly, the efficiency remains stable during several consecutive cycles, demonstrating a high adsorption capacity and a long lifetime and reusability of the membranes. The recovery of adsorbed precious metal ions converted into elemental metals is shown to be a general feature of these membranes, with platinum and silver successfully recovered from saturated hybrid membranes by a simple thermal reduction. The separation performance, evaluated on real electroplating industrial wastewater containing chromium and nickel, is found to exceed 99% at a permeability as high as 2.92 × 10–16 m2, that is, at least 4 orders of magnitude higher than typical nanofiltration membranes, conclusively validating the technology under stringent real conditions

Hybrid Amyloid Membranes for Continuous Flow Catalysis

Amyloid fibrils are promising nanomaterials for technological applications such as biosensors, tissue engineering, drug delivery, and optoelectronics. Here we show that amyloid–metal nanoparticle hybrids can be used both as efficient active materials for wet catalysis and as membranes for continuous flow catalysis applications. Initially, amyloid fibrils generated <i>in vitro</i> from the nontoxic β-lactoglobulin protein act as templates for the synthesis of gold and palladium metal nanoparticles from salt precursors. The resulting hybrids possess catalytic features as demonstrated by evaluating their activity in a model catalytic reaction in water, e.g., the reduction of 4-nitrophenol into 4-aminophenol, with the rate constant of the reduction increasing with the concentration of amyloid–nanoparticle hybrids. Importantly, the same nanoparticles adsorbed onto fibrils surface show improved catalytic efficiency compared to the same unattached particles, pointing at the important role played by the amyloid fibril templates. Then, filter membranes are prepared from the metal nanoparticle-decorated amyloid fibrils by vacuum filtration. The resulting membranes serve as efficient flow catalysis active materials, with a complete catalytic conversion achieved within a single flow passage of a feeding solution through the membrane

Twofold Light and Magnetic Responsive Behavior in Nanoparticle–Lyotropic Liquid Crystal Systems

We demonstrate the dual magnetic and light responsive nature of hybrid mesophases constituted by Fe₃O₄ nanoparticles dispersed in lipid-based lyotropic liquid crystals (LC). When subjected to an external magnetic field in the mesophase isotropic state, the nanoparticles aggregate and orient along the magnetic field direction, and upon cooling the system through the disorder–order transition the aggregates drive the orientation of the mesophase via heterogeneous nucleation; furthermore, order–disorder transitions in the lipidic mesophase can be triggered by Fe₃O₄-induced photothermal effect under visible light exposure. Both the orientational order and the photothermal effect of the hybrid mesophase can be tuned by the nanoparticle content, offering a general route for controlled assembly of complex fluids with combined magnetic and light responsiveness

Gelation, Phase Behavior, and Dynamics of β‑Lactoglobulin Amyloid Fibrils at Varying Concentrations and Ionic Strengths

We have investigated the thermodynamic and dynamic behavior of multistranded β-lactoglobulin protein fibrils in water, by combining static, dynamic, and depolarized dynamic light scattering (SLS, DLS, DDLS), small angle neutron scattering (SANS), rheology, and cryogenic transmission electron microscopy (cryo-TEM). We focus on the region of the phase diagram at which ionic strength and concentration changes induce transitions in gelation and lyotropic liquid crystalline behavior. An increase in ionic strength, induced by NaCl salt, progressively causes the phase transitions from nematic (N) to gel (G) phases; a further increase causes the transition to a translucent phase and to a macroscopic phase separation, respectively. An increase in fibril concentration induces first a phase transition from an isotropic (I) to a nematic phase (N); a further increase induces the formation of a gel phase. The protein gel strength is investigated by rheology measurements. SANS and osmotic compressibility calculated by SLS measurements clearly capture the main features of the IN transition of β-lactoglobulin protein fibrils. The form and structure factors measured by scattering experiments are analyzed by the polymer reference interaction site model (PRISM). Dynamics of the protein fibrils at different concentrations, measured by polarized and depolarized dynamic light scattering, show both individual and collective diffusion after the isotropic–nematic transition. Above this transition, cryo-TEM images further demonstrate the alignment of the protein fibrils, which is quantified by a 2D order parameter. This work discusses comprehensively, both experimentally and theoretically, the thermodynamics and dynamic features of β-lactoglobulin amyloid fibrils in a vast region of the concentration–ionic strength phase diagram

Magnetic-Responsive Hybrids of Fe₃O₄ Nanoparticles with β‑Lactoglobulin Amyloid Fibrils and Nanoclusters

We report on the synthesis and magnetic-responsive behavior of hybrids formed by dispersing negatively charged iron oxide (Fe₃O₄) magnetic nanoparticles in positively charged β-lactoglobulin protein solutions at acidic pH, followed by heating at high temperatures. Depending on the pH used, different hybrid aggregates can be obtained, such as nanoparticle-modified amyloid fibrils (pH 3) and spherical nanoclusters (pH 4.5). We investigate the effect of magnetic fields of varying strengths (0–5 T) on the alignment of these Fe₃O₄-modified amyloid fibrils and spherical nanoclusters using a combination of scattering, birefringence and microscopic techniques and we find a strong alignment of the hybrids upon increasing the intensity of the magnetic field, which we quantify <i>via</i> 2D and 3D order parameters. We also demonstrate the possibility of controlling magnetically the sol–gel behavior of these hybrids: addition of salt (NaCl, 150 mM) to a solution containing nanoparticles modified with β-lactoglobulin amyloid fibrils (2 wt % fibrils modified with 0.6 wt % Fe₃O₄ nanoparticles) induces first the formation of a reversible gel, which can then be converted back to solution upon application of a moderate magnetic field of 1.1 T. These hybrids offer a new appealing functional colloidal system in which the aggregation, orientational order and rheological behavior can be efficiently controlled in a purely noninvasive way by external magnetic fields of weak intensity

Switching the Morphologies of Cylindrical Polycation Brushes by Ionic and Supramolecular Inclusion Complexes

Switching the Morphologies of Cylindrical Polycation Brushes by Ionic and Supramolecular Inclusion Complexe

Reversible Aggregation of DNA-Decorated Gold Nanoparticles Controlled by Molecular Recognition

The programmable assembly of functional nanomaterials has been extensively addressed; however, their selective reversible assembly in response to an external stimulus has been more difficult to realize. The specificity and programmable interactions of DNA have been exploited for the rational self-assembly of DNA-conjugated nanoparticles, and here we demonstrate the sequence-controlled disaggregation of DNA-modified gold nanoparticles simply by employing two complementary oligonucleotides. Target oligonucleotides with perfectly matching sequence enabled dissociation of aggregated nanoparticles, whereas oligonucleotides differing by one nucleotide did not cause disassembly of the aggregated nanoparticles. Physical aspects of this process were characterized by UV–vis absorption, light scattering, and transmission electron microscopy. This strategy for programmed disassembly of gold nanoparticles in response to biological stimuli demonstrates a fundamentally important concept anticipated to be useful for diverse applications involving molecular recognition

Self-Assembly of Ovalbumin into Amyloid and Non-Amyloid Fibrils

We study the fibrillation pathway of ovalbumin protein and report the simultaneous formation of several types of fibrils, with clear structural and physical differences. We compare the fibrillation mechanisms at low pH with and without salt, and follow the kinetics of fibrils growth by atomic force microscopy (AFM), static and dynamic light scattering (SLS, DLS), and small-angle X-ray scattering (SAXS). We show that, among the morphologies identified, long semiflexible amyloid fibrils (type I), with persistence length Lp ∼ 3 μm, Young's modulus E ∼ 2.8 GPa, and cross-β structure are formed. We also observe much more flexible fibrils (type III, Lp ∼ 63 nm), that can assemble into multistranded ribbons with time. They show significantly lower intrinsic stiffness (1.1 GPa) and a secondary structure, which is not characteristic of the well-ordered amyloids, as determined by circular dichroism (CD), wide-angle X-ray scattering (WAXS), and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). In between these two main classes of fibrils, a third family, with intermediate flexibility (type II, Lp ∼ 300 nm), is also resolved

Complexation of β-Lactoglobulin Fibrils and Sulfated Polysaccharides

Fibrils of β-lactoglobulin, formed by heating at pH 2, were titrated with a sulfated polysaccharide (κ-carrageenan) to determine the morphology and mechanism of complex formation at low pH. Structural information on the resultant complexes was gathered using transmission electron microscopy, atomic force microscopy, Doppler electrophoresis, and small-angle neutron scattering. Electrophoresis demonstrated that the carrageenan complexed with protein fibrils until reaching a maximum complexation efficiency at a protein/polysaccharide (r) weight ratio of 5:3. Neutron scattering and microscopy indicated an increasing formation of spherical aggregates attached along the protein fibrils with increases in the carrageenan concentration. These globular particles had an average diameter of 30 nm. Small-angle neutron scattering of these complexes could be accurately described by a form factor corresponding to multistranded twisted ribbons with spherical aggregates along their contour length, arranged in a necklace configuration

Salt-Induced Aggregation of Polyelectrolyte−Amphiphilic Dendron Complexes in THF Solutions

Complexes were prepared by complexing amphiphilic dendrons (first, second, and third generations) with an oppositely charged polyelectrolyte through ionic interaction. Their chemical structure can be confirmed by nuclear magnetic resonance spectroscopy (1H NMR), Fourier transform infrared spectroscopy (FT-IR), and elemental analysis (EA). All of the complexes were investigated with respect to their aggregation behavior. Under the inducement of salt addition in THF, they showed interesting self-assembly behavior, forming micelles and vesicles depending on the amount of the salt. The formation of these self-assemblies has been proven by dynamic light scattering (DLS), static light scattering (SLS), and electron microscopy (EM) as well as atomic force microscopy (AFM). Dilution experiments showed that the formed vesicles have good stability against dilution as well as polyelectrolyte behavior. The formation of micelle and vesicle aggregates described in this article is probably based on the entropy effect and the shape transformation of building blocks caused by salt addition