Supramolecular chirality and crystallization from biocatalytic self-assembly in lipidic cubic mesophases

Biocatalytic self-assembly in a nanoconfined environment is widely used in nature to construct complex structures that endow special characteristics to life. There is tremendous interest in mimicking such bottom-up processes to fabricate functional materials. In this study, we have investigated a novel biomimetic scaffold based on lipidic cubic mesophases (LCMs), which provide a special nanoconfined environment for biocatalytic self-assembly and subsequent formation of organic crystals. (R)-Benzoin generated in situ from benzaldehyde in a reaction catalyzed by the enzyme benzaldehyde lyase (BAL) exhibits – when confined within LCMs – enhanced chirality compared to (R)-benzoin in solution or (R)-benzoin-doped LCMs. We infer that a metastable state is formed under kinetic control that displays enhanced supramolecular chirality. As they age, these metastable structures can further grow into thermodynamically stable crystals. The biomimetic, nanoconfined environment provided by the LCMs plays a key role in the development of supramolecular chirality and subsequent crystallization.ISSN:2040-3364ISSN:2040-337

Enzyme Kinetics in Liquid Crystalline Mesophases: Size Matters, But Also Topology

Lyotropic liquid crystalline systems (LLCs) are excellent immobilizing carriers for enzymes, due to their biocompatibility and well-defined pore nanostructure. Here we show that the liquid crystalline mesophase topology can greatly influence the enzymatic activity in a typical peroxidase (Horseradish peroxidase, HRP) enzymatic reaction. Enzyme kinetics was investigated in different LLC mesophases based on monolinolein, with varying symmetries and dimensions such as the 1D cylindrical inverse hexagonal phase (H_II), the 2D planar lamellar phase (L_α), and two 3D bicontinuous cubic phases of double diamond (Pn3m) and gyroid (Ia3d) space groups. As expected, the mesophase with largest water channel size shows highest activity, regardless of the topology. Interestingly, however, when mesophases with different topologies have the same water channel size, then the topology plays the dominant role, and the enzyme showed the highest activity in the 3D tetra-fold connected Pn3m, followed by the Ia3d with trifold connectivity, and finally the 1D H_II phase. This study demonstrates that the enzymatic activity in LLC mesophases depends on both the water channel size and the topology of the mesophase

Controlling Anisotropic Drug Diffusion in Lipid-Fe₃O₄ Nanoparticle Hybrid Mesophases by Magnetic Alignment

We present a new strategy to control the anisotropic diffusion of hydrophilic drugs in lyotropic liquid crystals via the dispersion of magnetic nanoparticles in the mesophase, followed by reorientation of the mesophase domains via an external magnetic field. We select a lipid reverse hexagonal phase doped with magnetic iron oxide nanoparticles and glucose and caffeine as model hybrid mesophase and hydrophilic drugs, respectively. Upon cooling through the disorder–order phase transition of the hexagonal phase and under exposure to an external moderate magnetic field (1.1 T), both the nanoparticles and the hexagonal domains align with their columnar axes along the field direction. As a result, the water nanochannels of the inverted hexagonal domains also align parallel to the field direction, leading to a drug diffusion coefficient parallel to the field direction much larger than what was measured perpendicularly: in the case of glucose, for example, this difference in diffusion coefficients approaches 1 order of magnitude. Drug diffusion of the unaligned reverse hexagonal phase, which consists of randomly distributed domains, shows values in between the parallel and transversal diffusion values. This study shows that modifying the overall alignment of anisotropic mesophases via moderate external fields is a valuable means to control the corresponding transport tensor of the mesophase and demonstrates that the orientation of the domains plays an important role in the diffusion process of foreign hydrophilic molecules

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

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

Lyotropic Liquid Crystalline Cubic Phases as Versatile Host Matrices for Membrane-Bound Enzymes

Lyotropic liquid crystalline cubic mesophases can function as host matrices for enzymes because of their biomimetic structural characteristics, optical transparency, and capability to coexist with water. This study demonstrates that the in meso immobilized membrane-bound enzyme d-fructose dehydrogenase (FDH) preserves its full activity, follows ideal Michaelis–Menten kinetics, and shows improved stability compared to its behavior in solution. Even after 5 days, the immobilized FDH retained its full activity in meso, whereas a model hydrophilic enzyme, horseradish peroxidase, maintained only 21% of its original activity. We reason that the lipidic bilayers in the three-dimensional structures of cubic mesophases provide an ideal environment for the reconstitution of a membrane-bound enzyme. The preserved activity, long-term stability, and reusability demonstrate that these hybrid nanomaterials are ideal matrices for biosensing and biocatalytic fuel cell applications