Non-mechanical nano- and microscale pumps that function without the aid of an external power source and provide precise control over the flow rate in response to specific signals are needed for the development of new autonomous nano- and microscale systems. Here we show that surface-immobilized enzymes that are independent of adenosine triphosphate function as self-powered micropumps in the presence of their respective substrates. In the four cases studied (catalase, lipase, urease and glucose oxidase), the flow is driven by a gradient in fluid density generated by the enzymatic reaction. The pumping velocity increases with increasing substrate concentration and reaction rate. These rechargeable pumps can be triggered by the presence of specific analytes, which enables the design of enzyme-based devices that act both as sensor and pump. Finally, we show proof-of-concept enzyme-powered devices that autonomously deliver small molecules and proteins in response to specific chemical stimuli, including the release of insulin in response to glucose.

Surface-bound enzymes can act as pumps that drive large-scale fluid flows in the presence of their substrates or promoters. Thus, enzymatic catalysis can be harnessed for "on demand" pumping in nano-and microfluidic devices powered by an intrinsic energy source. The mechanisms controlling the pumping have not, however, been completely elucidated. Herein, we combine theory and experiments to demonstrate a previously unreported spatiotemporal variation in pumping behavior in urease-based pumps and uncover the mechanisms behind these dynamics. We developed a theoretical model for the transduction of chemical energy into mechanical fluid flow in these systems, capturing buoyancy effects due to the solution containing nonuniform concentrations of substrate and product. We find that the qualitative features of the flow depend on the ratios of diffusivities δ = D P =D S and expansion coefficients β = β P =β S of the reaction substrate (S) and product (P). If δ > 1 and δ > β (or if δ < 1 and δ < β), an unexpected phenomenon arises: the flow direction reverses with time and distance from the pump. Our experimental results are in qualitative agreement with the model and show that both the speed and direction of fluid pumping (i) depend on the enzyme activity and coverage, (ii) vary with the distance from the pump, and (iii) evolve with time. These findings permit the rational design of enzymatic pumps that accurately control the direction and speed of fluid flow without external power sources, enabling effective, self-powered fluidic devices.enzyme micropumps | solutal convection | flow reversal | self-powered systems | catalysis N onmechanical nano/microscale pumps that provide precise control over fluid flow without an external power source and are capable of turning on in response to specific analytes in solution are needed for the next generation of smart micro-and nanoscale devices. Specific applications involve neutralization of toxins (1), on-demand drug or antidote delivery (2), and the directed focusing of targeted analytes for optimal sensing (3). Recent experiments have shown that surface-bound enzymes act as pumps in the presence of their specific substrates or promoters, driving large-scale fluid flows (2, 4). Furthermore, fluid velocity increases with increasing reaction rate. Therefore, enzymatic catalysis provides an intrinsic energy source for fluid movement and thus can be harnessed to overcome a significant obstacle in nano-and microfluidics: the need to use pressuredriven pumps to push fluids through devices. To fully exploit this behavior, it is vital to develop a fundamental understanding of how the chemical energy is transduced into mechanical fluid flow. One possible mechanism involves thermal buoyancy effects: An exothermic reaction catalyzed by the enzyme lowers the solution density, causing fluid to be drawn in and rise above the pump (Fig. 1A). Not all observed pumping, however, can be explained by this mechanism. In several instances (4), the thermal input from the enzymatic reaction is far lo...

Immobilized enzymes generate net fluid flow when exposed to specific reagents in solution. Thus, they function as self-powered platforms that combine sensing and on-demand fluid pumping. To uncover the mechanism of pumping, we examine the effects of solutal and thermal buoyancy on the behavior of phosphatase-based micropumps, using a series of reactants with known thermodynamic and kinetic parameters. By combining modeling and experiments, we perform the first quantitative comparison of thermal and solutal effects in an enzyme micropump system. Despite the significant exothermicity of the catalyzed reactions, we find that thermal effects play a minimal role in the observed fluid flow. Instead, fluid transport in phosphatase micropumps is governed by the density difference between the reactants and the products of the reaction. This surprising conclusion suggests new design principles for catalytic pumps.

The directed transport of microparticles in microfluidic devices is vital for efficient bioassays and fabrication of complex microstructures. There remains, however, a need for methods to propel and steer microscopic cargo that do not require modifying these particles. Using theory and experiments, we show that catalytic surface reactions can be used to deliver microparticle cargo to specified regions in microchambers. Here reagents diffuse from a gel reservoir and react with the catalyst-coated surface. Fluid density gradients due to the spatially varying reagent concentration induce a convective flow, which carries the suspended particles until the reagents are consumed. Consequently, the cargo is deposited around a specific position on the surface. The velocity and final peak location of the cargo can be tuned independently. By increasing the local particle concentration, highly sensitive assays can be performed efficiently and rapidly. Moreover, the process can be repeated by introducing fresh reagent into the microchamber.

Rapid, easy‐to‐use, and portable devices that can provide a read‐out without the need for expensive equipment represent the future of sensing technology, with applications in areas like environmental, food, chemical, and biological safety. Enzymes immobilized on a surface function as micropumps in the presence of species (e.g., substrate, cofactor, or biomarker) that trigger the enzymatic reaction. The flow speed in these devices increases with increasing reaction rate. This allows the detection of substances that inhibit the enzymatic reaction. Using this principle, sensors for toxic substances, like mercury, cadmium, cyanide, and azide, were designed using urease and catalase‐powered pumps, respectively, with limits of detection well below the concentrations permitted by the Environmental Protection Agency. The study was also extended to other inhibitors for these enzymes. The sensing range of fluid flow‐based inhibitor assays depends on the type of inhibition, the enzyme concentration on the sensing platform, and, for competitive inhibition, the concentration of substrate used.

ConspectusAutonomous micro- and nanoscale systems have revolutionized the way scientists look into the future, opening up new frontiers to approach and solve problems via a more bioinspired route. However, to achieve systems with higher complexity, superior output control, and multifunctionality, an in-depth study of the different factors that affect micro- and nanomotor behavior is crucial. From a fundamental perspective, the mechanical response of micro- and nanomotors still requires further study in order to have a better understanding of how exactly these systems operate and the different mechanisms of motion that can be combined into one system to achieve an optimal response. From a design engineering point of view, compatibility, degradability, specificity, sensitivity, responsiveness, and efficiency of the active systems fabricated to this point have to be addressed, with respect to the potential of these devices for biomedical applications. Nonetheless, optimizing the system with regards to all these areas is a challenging task with the micro- and nanomotors studied to date, as most of them consist of materials or designs that are unfavorable for further chemical or physical manipulation.As this new field of self-powered systems moves forward, the need for motor prototypes with different sizes, shapes, chemical functionalities, and architectures becomes increasingly important and will define not only the way active systems are powered, but also the methods for motor fabrication. Bottom-up supramolecular approaches have recently emerged as great candidates for the development of active structures that allow for chemical or physical functionalization, shape transformation, and compartmentalization, in a structure that provides a soft interface to improve molecular recognition and cell uptake.Our group pioneers the use of supramolecular structures as catalytically propelled systems via the fabrication of stomatocyte or tubular-shaped motors capable of displaying active motion in a substrate concentration-dependent fashion. This behavior demonstrates the potential of bottom-up assemblies for powering motion at the micro- or nanoscale, with a system that can be readily tuned and controlled at the molecular level. In this Account, we highlight the steps we have taken in order to understand and optimize the design of catalytically powered polymersome-based motors. Our research has been focused on addressing the importance of motor architecture, motion activation, direction control, and biological integration. While our work supports the feasibility of supramolecular structures for the design of active systems, we strongly believe that we are still in the initial stages of unveiling the full potential of supramolecular chemistry in the micro- and nanomotor field. We look forward to using this approach for the development of multifunctional and stimuli-responsive systems in the near future.