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.

Recent experimental results have shown that active enzymes can diffuse faster when they are in the presence of their substrates. Fluorescence correlation spectroscopy (FCS), which relies on analyzing the fluctuations in fluorescence intensity signal to measure the diffusion coefficient of particles, has typically been employed in most of the prior studies. However, flaws in the FCS method, due to its high sensitivity to the environment, have recently been evaluated, calling the prior diffusion results into question. It behooves us to adopt complementary and direct methods to measure the mobility of enzymes in solution. Herein, we use a novel technique of direct single-molecule imaging to observe the diffusion of single enzymes. This technique is less sensitive to intensity fluctuations and gives the diffusion coefficient directly based on the trajectory of the enzymes. Our measurements recapitulate that enzyme diffusion is enhanced in the presence of its substrate and find that the relative increase in diffusion of a single enzyme is even higher than those previously reported using FCS. We also use this complimentary method to test if the total enzyme concentration affects the relative increase in diffusion and if enzyme oligomerization state changes during catalytic turnover. We find that the diffusion increase is independent of the total background concentration of enzyme and the catalysis of substrate does not change the oligomerization state of enzymes.

The environment is affected by the actions of mankind in multitudinous ways, many of which are detrimental, giving rise to pollution and toxic waste, ultimately making our planet less inhabitable. While remediation and new regulations help to prevent pollution and toxic waste, there is also a need to change the behavior of future generations of consumers and producers of new products. Future chemists and innovators are charged with the responsibility of developing new chemical processes and products that not only meet the needs of our growing population (in terms of energy, clean water and food), but also protect human health and the environment. Green Chemistry is a revolution in the design of molecules that provides new opportunities for economic development while considering the impact on health and the environment. Green Chemistry utilizes a set of guiding principles, originally provided by Anastas and Warner (Green Chemistry: Theory and Practice, 1998), aimed at decreasing/removing the use/generation of hazardous substances in the design, manufacture and application of products. To help improve the creative and innovative thinking behind Green Chemistry, it is important to expose chemistry students to these principles at the undergraduate level. While suitable Green Chemistry experiments are known, successful implementation requires running test trials and performing additional basic research. We have embarked upon the testing and further design of experiments for implementation into an undergraduate laboratory course and report our results in this endeavor.

No abstract