The self-assembly of block copolymers into 1D, 2D and 3D nano- and microstructures is of great interest for a wide range of applications. A key challenge in this field is obtaining independent ...control over molecular structure and hierarchical structure in all dimensions using scalable one-pot chemistry. Here we report on the ring opening polymerization-induced crystallization-driven self-assembly (ROPI-CDSA) of poly-L-lactide-block-polyethylene glycol block copolymers into 1D, 2D and 3D nanostructures. A key feature of ROPI-CDSA is that the polymerization time is much shorter than the self-assembly relaxation time, resulting in a non-equilibrium self-assembly process. The self-assembly mechanism is analyzed by cryo-transmission electron microscopy, wide-angle x-ray scattering, Fourier transform infrared spectroscopy, and turbidity studies. The analysis revealed that the self-assembly mechanism is dependent on both the polymer molecular structure and concentration. Knowledge of the self-assembly mechanism enabled the kinetic trapping of multiple hierarchical structures from a single block copolymer.
The virus like particle (VLP) derived from bacteriophage P22 presents a unique platform for constructing catalytically functional nanomaterials by encapsulation of enzymes into its interior. ...Encapsulation has been engineered to be genetically programmed allowing “one pot” synthesis and incorporation of desired enzymes. The unique characteristic that separates P22 from other VLP systems is the ability to modulate the overall volume and porosity of the VLP structure, thus controlling substrate access to the encapsulated enzyme. The present study demonstrates incorporation of an enzyme, alcohol dehydrogenase D, with the highest internal loading for an active enzyme by any VLP described thus far. In addition, we show that not only does encapsulating AdhD inside P22 affect its kinetic parameters in comparison with the “free” enzyme, but transformation of P22 to different morphological states, which changes the internal volume of the VLP, yields changes in the overall activity of the encapsulated enzyme as well. The findings reported here clearly illustrate that P22 holds potential for synthetic approaches to create nanoreactors, by design, using the power of highly evolved enzymes for chemical transformations.
Protein–metal–organic frameworks (p-MOFs) are a prototypical example of how synthetic biological hybrid systems can be used to develop next-generation materials. Controlling p-MOF formation enables ...the design of hybrid materials with enhanced biological activity and high stability. However, such control is yet to be fully realized due to an insufficient understanding of the governing nucleation and growth mechanisms in p-MOF systems. The structural evolution of p-MOFs was probed by cryo-transmission electron microscopy, revealing nonclassical pathways via dissolution–recrystallization of highly hydrated amorphous particles and solid-state transformation of a protein-rich amorphous phase. On the basis of these data, we propose a general description of p-MOF crystallization which is best characterized by particle aggregation and colloidal theory for future synthetic strategies.
There has been much interest in the construction of soft nanomaterials in solution due to a desire to emulate the exquisite structure and function of Nature's equivalents (e.g. enzymes, viruses, ...proteins and DNA). Nature's soft nanomaterials are capable of selectivity, precision and efficiency in areas such as information storage and replication, transportation and delivery, and synthesis and catalysis. To this end, the use of small molecules, amphiphiles, colloids, and polymers have been investigated for the development of advanced materials in myriad fields of biomedicine and synthetic chemistry. Two major challenges are faced in this area of research: the reproducible, scalable and precise synthesis of such constructs and the reliable, accurate and in-depth analysis of these materials. This tutorial review will focus on this second aspect and provide a guide for the characterisation and analysis of soft nanomaterials in solution using scattering and microscopic techniques.
Innovations in liquid‐phase electron microscopy (LP‐EM) have made it possible to perform experiments at the optimized conditions needed to examine soft matter. The main obstacle is conducting ...experiments in such a way that electron beam radiation can be used to obtain answers for scientific questions without changing the structure and (bio)chemical processes in the sample due to the influence of the radiation. By overcoming these experimental difficulties at least partially, LP‐EM has evolved into a new microscopy method with nanometer spatial resolution and sub‐second temporal resolution for analysis of soft matter in materials science and biology. Both experimental design and applications of LP‐EM for soft matter materials science and biological research are reviewed, and a perspective of possible future directions is given.
Liquid‐phase electron microscopy (LP‐EM) provides nanometer spatial resolution and sub‐second temporal resolution for analysis of soft matter. The experimental design has to optimize resolution versus radiation damage. Both techniques and applications of LP‐EM for soft matter materials science and biological research are reviewed, and a perspective of possible future directions is given.
Liquid cell transmission electron microscopy (LCTEM) can provide direct observations of solution-phase nanoscale materials, and holds great promise as a tool for monitoring dynamic self-assembled ...nanomaterials. Control over particle behavior within the liquid cell, and under electron beam irradiation, is of paramount importance for this technique to contribute to our understanding of chemistry and materials science at the nanoscale. However, this type of control has not been demonstrated for complex, organic macromolecular materials, which form the basis for all biological systems and all of polymer science, and encompass important classes of advanced porous materials. Here we show that by controlling the liquid cell membrane surface chemistry and electron beam conditions, the dynamics and growth of metal–organic frameworks (MOFs) can be observed. Our results demonstrate that hybrid organic/inorganic beam-sensitive materials can be analyzed with LCTEM and, at least in the case of ZIF-8 dynamics, the results correlate with observations from bulk growth or other standard synthetic conditions. Furthermore, we show that LCTEM can be used to better understand how changes to synthetic conditions result in changes to particle size. We anticipate that direct, nanoscale imaging by LCTEM of MOF nucleation and growth mechanisms may provide insight into controlled MOF crystal morphology, domain composition, and processes influencing defect formation.
Developing methods for investigating coupled enzyme systems under conditions that mimic the cellular environment remains a significant challenge. Here we describe a biomimetic approach for ...constructing densely packed and confined multienzyme systems through the co-encapsulation of 2 and 3 enzymes within a virus-like particle (VLP) that perform a coupled cascade of reactions, creating a synthetic metabolon. Enzymes are efficiently encapsulated in vivo with known stoichiometries, and the kinetic parameters of the individual and coupled activities are characterized. From the results we develop and validate a mathematical model for predicting the expected kinetics for coupled reactions under co-localized conditions.
The chemistry of highly evolved protein-based compartments has inspired the design of new catalytically active materials that self-assemble from biological components. A frontier of this biodesign is ...the potential to contribute new catalytic systems for the production of sustainable fuels, such as hydrogen. Here, we show the encapsulation and protection of an active hydrogen-producing and oxygen-tolerant NiFe-hydrogenase, sequestered within the capsid of the bacteriophage P22 through directed self-assembly. We co-opted Escherichia coli for biomolecular synthesis and assembly of this nanomaterial by expressing and maturing the EcHyd-1 hydrogenase prior to expression of the P22 coat protein, which subsequently self assembles. By probing the infrared spectroscopic signatures and catalytic activity of the engineered material, we demonstrate that the capsid provides stability and protection to the hydrogenase cargo. These results illustrate how combining biological function with directed supramolecular self-assembly can be used to create new materials for sustainable catalysis.
Amphiphilic small molecules and polymers form commonplace nanoscale macromolecular compartments and bilayers, and as such are truly essential components in all cells and in many cellular processes. ...The nature of these architectures, including their formation, phase changes, and stimuli-response behaviors, is necessary for the most basic functions of life, and over the past half-century, these natural micellar structures have inspired a vast diversity of industrial products, from biomedicines to detergents, lubricants, and coatings. The importance of these materials and their ubiquity have made them the subject of intense investigation regarding their nanoscale dynamics with increasing interest in obtaining sufficient temporal and spatial resolution to directly observe nanoscale processes. However, the vast majority of experimental methods involve either bulk-averaging techniques including light, neutron, and X-ray scattering, or are static in nature including even the most advanced cryogenic transmission electron microscopy techniques. Here, we employ in situ liquid-cell transmission electron microscopy (LCTEM) to directly observe the evolution of individual amphiphilic block copolymer micellar nanoparticles in solution, in real time with nanometer spatial resolution. These observations, made on a proof-of-concept bioconjugate polymer amphiphile, revealed growth and evolution occurring by unimer addition processes and by particle–particle collision-and-fusion events. The experimental approach, combining direct LCTEM observation, quantitative analysis of LCTEM data, and correlated in silico simulations, provides a unique view of solvated soft matter nanoassemblies as they morph and evolve in time and space, enabling us to capture these phenomena in solution.
Cetacean morbillivirus (CeMV) has caused several epizootics in multiple species of cetaceans globally and is an emerging disease among cetaceans in Australia. We detected CeMV in 2 stranded coastal ...Indo-Pacific bottlenose dolphins (Tursiops aduncus) in Western Australia. Preliminary phylogenetic data suggest that this virus variant is divergent from known strains.
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