Polyethylene Glycol Crowder's Effect on Enzyme Aggregation, Thermal Stability, and Residual Catalytic Activity.

Protein stability and performance in various natural and artificial systems incorporating many other macromolecules for therapeutic, diagnostic, sensor, and biotechnological applications attract increasing interest with the expansion of these technologies. Here we address the catalytic activity of lysozyme protein (LYZ) in the presence of a polyethylene glycol (PEG) crowder in a broad range of concentrations and temperatures in aqueous solutions of two different molecular mass PEG samples (Mw = 3350 and 10000 g/mol). The phase behavior of PEG-protein solutions is examined by using dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS), while the enzyme denaturing is monitored by using an activity assay (AS) and circular dichroism (CD) spectroscopy. Molecular dynamic (MD) simulations are used to illustrate the effect of PEG concentration on protein stability at high temperatures. The results demonstrate that LYZ residual activity after 1 h incubation at 80 °C is improved from 15% up to 55% with the addition of PEG. The improvement is attributed to two underlying mechanisms. (i) Primarily, the stabilizing effect is due to the suppression of the enzyme aggregation because of the stronger PEG-protein interactions caused by the increased hydrophobicity of PEG and lysozyme at elevated temperatures. (ii) The MD simulations showed that the addition of PEG to some degree stabilizes the secondary structures of the enzyme by delaying unfolding at elevated temperatures. The more pronounced effect is observed with an increase in PEG concentration. This trend is consistent with CD and AS experimental results, where the thermal stability is strengthened with increasing of PEG concentration and molecular mass. The results show that the highest stabilizing effect is approached at the critical overlap concentration of PEG.

Designing Highly Thermostable Lysozyme-Copolymer Conjugates: Focus on Effect of Polymer Concentration.

Designing biomaterials capable of functioning in harsh environments is vital for a range of applications. Using molecular dynamics simulations, we show that conjugating lysozymes with a copolymer [poly(GMA- stat-OEGMA)] comprising glycidyl methacrylate (GMA) and oligo(ethylene glycol) methyl ether methacrylate (OEGMA) results in a dramatic increase of stability of these enzymes at high temperatures provided that the concentration of the copolymer in the close vicinity of the enzyme exceeds a critical value. In our simulations, we use triads containing the same ratio of GMA to OEGMA units as in our recent experiments (N. S. Yadavalli et al., ACS Catalysis, 2017, 7, 8675). We focus on the dynamics of the conjugate at high temperatures and on its structural stability as a function of the copolymer/water content in the vicinity of the enzyme. We show that the dynamics of phase separation in the water-copolymer mixture surrounding the enzyme is critical for the structural stability of the enzyme. Specifically, restricting water access promotes the structural stability of the lysozyme at high temperatures. We identified critical water concentration below which we observe a robust stabilization; the phase separation is no longer observed at this low fraction of water so that the water domains promoting unfolding are no longer formed in the vicinity of the enzyme. This understanding provides a basis for future studies on designing a range of enzyme-copolymer conjugates with improved stability.

Chemo-responsive, self-oscillating gels that undergo biomimetic communication.

Species ranging from single-cell organisms to social insects can undergo auto-chemotaxis, where the entities move towards a chemo-attractant that they themselves emit. Polymer gels undergoing the self-oscillating Belousov-Zhabotinsky (BZ) reaction exhibit autonomous, periodic pulsations, which produce chemical species collectively referred to as the activator. The diffusion of this activator into the surrounding solution affects the dynamic behavior of neighboring BZ gels and hence, the BZ gels not only emit, but also respond to self-generated chemical gradients. This review describes recent experimental and computational studies that reveal how this biomimetic behavior effectively allows neighboring BZ gels to undergo cooperative, self-propelled motion. These distinctive properties of the BZ gels provide a route for creating reconfigurable materials that autonomously communicate with neighboring units and thereby actively participate in constructing the desired structures.

Harnessing interfacially-active nanorods to regenerate severed polymer gels.

With newly developed computational approaches, we design a nanocomposite that enables self-regeneration of the gel matrix when a significant portion of the material is severed. The cut instigates the dynamic cascade of cooperative events leading to the regrowth. Specifically, functionalized nanorods localize at the new interface and initiate atom transfer radical polymerization with monomers and cross-linkers in the outer solution. The reaction propagates to form a new cross-linked gel, which can be tuned to resemble the uncut material.

Controlling Degradation and Erosion of Polymer Networks: Insights from Mesoscale Modeling.

Understanding and controlling degradation of polymer networks on the mesoscale is critical for a range of applications. We utilize dissipative particle dynamics to capture photocontrolled degradation and erosion processes in hydrogels formed by end-linking of four-arm polyethylene glycol precursors. We demonstrate that the polydispersity and the fraction of broken-off fragments scale with the relative extent of reaction. The reverse gel point measured is close to the value predicted by the bond percolation theory on a diamond lattice. We characterize the erosion process via tracking the mass loss that accounts for the fragments remaining in contact with the percolated network. We quantify the dependence of the mass loss on the extent of reaction and on the properties of the film prior to degradation. These results elucidate the main features of degradation and erosion on the mesoscale and could provide guidelines for future design of degrading materials with dynamically controlled properties.

Designing polymer gels and composites that undergo bio-inspired phototactic reconfiguration and motion.

Inspired by the adaptive behavior of photo-responsive biological organisms, we develop analytical and computational models to design polymer gels and composites that can be dynamically reconfigured and driven to move with the application of light. We focus on gels formed from poly(N-isopropylacrylamide) and functionalized with spirobenzopyran (SP) chromophores, which become hydrophobic under blue light in acidic aqueous solution. Using our modeling approaches, we irradiate the gels through photomasks and demonstrate that the shapes of the samples can be reversibly and remotely 'remolded' by varying the apertures in the masks. By simulating the effect of repeatedly moving the light across the sample, we also show that the gel can undergo directed motion. We then examine gels that contain both SP chromophores and the ruthenium catalysts that drive the oscillatory Belousov-Zhabotinsky reaction. These dual-functionalized gels undergo spontaneous, self-sustained motion even when the lights are held stationary. We also simulate the behavior of composites formed from SP-functionalized fibers embedded in the poly(N-isopropylacrylamide) gel. With the SP-functionalization confined to the fibers, light and heat act as orthogonal stimuli and thus the composites display distinctly different modes of movement when the different cues are applied to the samples. Overall, our findings provide guidelines for using light to controllably reconfigure the shape and drive the movement of gel-based materials and thus, tailor the material to display different functionalities.

Designing Dual-functionalized Gels for Self-reconfiguration and Autonomous Motion.

Human motion is enabled by the concerted expansion and contraction of interconnected muscles that are powered by inherent biochemical reactions. One of the challenges in the field of biomimicry is eliciting this form of motion from purely synthetic materials, which typically do not generate internalized reactions to drive mechanical action. Moreover, for practical applications, this bio-inspired motion must be readily controllable. Herein, we develop a computational model to design a new class of polymer gels where structural reconfigurations and internalized reactions are intimately linked to produce autonomous motion, which can be directed with light. These gels contain both spirobenzopyran (SP) chromophores and the ruthenium catalysts that drive the oscillatory Belousov-Zhabotinsky (BZ) reaction. Importantly, both the SP moieties and the BZ reaction are photosensitive. When these dual-functionalized gels are exposed to non-uniform illumination, the localized contraction of the gel (due to the SP moieties) in the presence of traveling chemical waves (due to the BZ reaction) leads to new forms of spontaneous, self-sustained movement, which cannot be achieved by either of the mono-functionalized networks.

Modeling chemoresponsive polymer gels.

Stimuli-responsive gels are vital components in the next generation of smart devices, which can sense and dynamically respond to changes in the local environment and thereby exhibit more autonomous functionality. We describe recently developed computational methods for simulating the properties of such stimuli-responsive gels in the presence of optical, chemical, and thermal gradients. Using these models, we determine how to harness light to drive shape changes and directed motion in spirobenzopyran-containing gels. Focusing on oscillating gels undergoing the Belousov-Zhabotinksy reaction, we demonstrate that these materials can spontaneously form self-rotating assemblies, or pinwheels. Finally, we model temperature-sensitive gels that encompass chemically reactive filaments to optimize the performance of this system as a homeostatic device for regulating temperature. These studies could facilitate the development of soft robots that autonomously interconvert chemical and mechanical energy and thus perform vital functions without the continuous need of external power sources.

Modeling the self-assembly of lipids and nanotubes in solution: forming vesicles and bicelles with transmembrane nanotube channels.

Via dissipative particle dynamics (DPD), we simulate the self-assembly of end-functionalized, amphiphilic nanotubes and lipids in a hydrophilic solvent. Each nanotube encompasses a hydrophobic stalk and two hydrophilic ends, which are functionalized with end-tethered chains. With a relatively low number of the nanotubes in solution, the components self-assemble into stable lipid-nanotube vesicles. As the number of nanotubes is increased, the system exhibits a vesicle-to-bicelle transition, resulting in stable hybrid bicelle. Moreover, our results reveal that the nanotubes cluster into distinct tripod-like structures within the vesicles and aggregate into a ring-like assembly within the bicelles. For both the vesicles and bicelles, the nanotubes assume trans-membrane orientations, with the tethered hairs extending into the surrounding solution or the encapsulated fluid. Thus, the hairs provide a means of regulating the transport of species through the self-assembled structures. Our findings provide guidelines for creating nanotube clusters with distinctive morphologies that might be difficult to achieve through more conventional means. The results also yield design rules for creating synthetic cell-like objects or microreactors that can exhibit biomimetic functionality.

Using light to guide the self-sustained motion of active gels.

We undertake the first computational study to determine the effect of light on polymer gels undergoing the Belousov-Zhabotinsky (BZ) reaction. The BZ gels are unique materials because they can undergo rhythmic mechanical oscillations in the absence of external stimuli. The BZ reaction, however, is photosensitive. Via simulations, we demonstrate that the interplay between the chemoresponsive gels and the photosensitive reaction can cause millimeter sized BZ gels to exhibit autonomous, directed motion or reorientation away from 4 the light. In effect, we show that these synthetic BZ "worms" display a fundamental biomimetic behavior: movement away from an adverse environmental condition, which in the context of the BZ reaction is the presence of light.

Spatial confinement controls self-oscillations in polymer gels undergoing the Belousov-Zhabotinsky reaction.

Chemoresponsive gels undergoing the Belousov-Zhabotinsky (BZ) reaction exhibit self-sustained pulsations, which can be harnessed to perform mechanical work. In technological applications, the gels would typically be confined between hard surfaces and thus, it is essential to establish how confinement affects these distinctive oscillations. Using theory and simulation, we pinpoint regions in phase space where the dynamic behavior of BZ gels critically depends on the presence of confining walls. We then illustrate how the wave propagation within thin samples can be tailored by selectively introducing "cut outs" in the bounding surfaces. The oscillations in the latter films are localized in specified areas, so the system contains well-defined oscillatory and nonoscillatory regions. The cut outs provide an effective means of tuning the mechanical action within the film and provide a route for tailoring the functionality of the material.

Gradient sensing in reactive, ternary membranes.

Using computer simulations, we investigate the behavior of reactive ternary ABC membranes that are subjected to an external, spatially nonuniform stimulus, which controls the rate of interconversion between the A and B components. We assume that A and B have different spontaneous curvatures. Furthermore, the C component is taken to be nonreactive and incompatible with both A and B. We find that a gradient in the applied stimulus causes the dynamic reconstruction of the membrane, with a preferential reorientation of the reactive AB domains along the gradient. In addition, the external gradient effectively controls the transport of the nonreactive C component within the membrane. The latter effect could potentially be exploited for cleaning the membrane of the nonreactive C "impurities" or for the targeted delivery of the C component to specific locations.