Staff member publications
Arqué, Xavier, Romero-Rivera, Adrian, Feixas, Ferran, Patiño, Tania, Osuna, Sílvia, SÃ¡nchez, Samuel, (2019). Intrinsic enzymatic properties modulate the self-propulsion of micromotors Nature Communications 10, (1), 2826
Bio-catalytic micro- and nanomotors self-propel by the enzymatic conversion of substrates into products. Despite the advances in the field, the fundamental aspects underlying enzyme-powered self-propulsion have rarely been studied. In this work, we select four enzymes (urease, acetylcholinesterase, glucose oxidase, and aldolase) to be attached on silica microcapsules and study how their turnover number and conformational dynamics affect the self-propulsion, combining both an experimental and molecular dynamics simulations approach. Urease and acetylcholinesterase, the enzymes with higher catalytic rates, are the only enzymes capable of producing active motion. Molecular dynamics simulations reveal that urease and acetylcholinesterase display the highest degree of flexibility near the active site, which could play a role on the catalytic process. We experimentally assess this hypothesis for urease micromotors through competitive inhibition (acetohydroxamic acid) and increasing enzyme rigidity (β-mercaptoethanol). We conclude that the conformational changes are a precondition of urease catalysis, which is essential to generate self-propulsion.
Keywords: Biocatalysis, Immobilized enzymes, Molecular machines and motors
Patiño, Tania, Arqué, Xavier, Mestre, Rafael, Palacios, Lucas, Sánchez, Samuel, (2018). Fundamental aspects of enzyme-powered micro- and nanoswimmers Accounts of Chemical Research 51, (11), 2662–2671
ConspectusSelf-propulsion at the nanoscale constitutes a challenge due to the need for overcoming viscous forces and Brownian motion. Inspired by nature, artificial micro- and nanomachines powered by catalytic reactions have been developed. Due to the toxicity of the most commonly used fuels, enzyme catalysis has emerged as a versatile and biocompatible alternative to generate self-propulsion. Different swimmer sizes, ranging from the nanoscale to the microscale, and geometries, including tubular and spherical shapes, have been explored. However, there is still a lack of understanding of the mechanisms underlying enzyme-mediated propulsion. Size, shape, enzyme quantity and distribution, as well as the intrinsic enzymatic properties, may play crucial roles in motion dynamics.In this Account, we present the efforts carried out by our group and others by the community on the use of enzymes to power micro- and nanoswimmers. We examine the different structures, materials, and enzymes reported so far to fabricate biocatalytic micro- and nanoswimmers with special emphasis on their effect in motion dynamics. We discuss the development of tubular micro- and nanojets, focusing on the different fabrication methods and the effect of length and enzyme localization on their motion behavior. In the case of spherical swimmers, we highlight the role of asymmetry in enzyme coverage and how it can affect their motion dynamics. Different approaches have been described to generate asymmetric distribution of enzymes, namely, Janus particles, polymeric vesicles, and non-Janus particles with patch-like enzyme distribution that we recently reported. We also examine the correlation between enzyme kinetics and active motion. Enzyme activity, and consequently speed, can be modulated by modifying substrate concentration or adding specific inhibitors. Finally, we review the theory of active Brownian motion and how the size of the particles can influence the analysis of the results. Fundamentally, nanoscaled swimmers are more affected by Brownian fluctuations than microsized swimmers, and therefore, their motion is presented as an enhanced diffusion with respect to the passive case. Microswimmers, however, can overcome these fluctuations and show propulsive or ballistic trajectories. We provide some considerations on how to analyze the motion of these swimmers from an experimental point of view. Despite the rapid progress in enzyme-based micro- and nanoswimmers, deeper understanding of the mechanisms of motion is needed, and further efforts should be aimed to study their lifetime, long-term stability, and ability to navigate in complex media.
Patiño, Tania, Feiner-Gracia, Natalia, Arqué, Xavier, Miguel-López, Albert, Jannasch, Anita, Stumpp, Tom, Schäffer, Erik, Albertazzi, Lorenzo, Sánchez, Samuel, (2018). Influence of enzyme quantity and distribution on the self-propulsion of non-Janus urease-powered micromotors Journal of the American Chemical Society 140, (25), 7896-7903
The use of enzyme catalysis to power micro- and nanomachines offers unique features such as biocompatibility, versatility, and fuel bioavailability. Yet, the key parameters underlying the motion behavior of enzyme-powered motors are not completely understood. Here, we investigate the role of enzyme distribution and quantity on the generation of active motion. Two different micromotor architectures based on either polystyrene (PS) or polystyrene coated with a rough silicon dioxide shell (PS@SiO2) were explored. A directional propulsion with higher speed was observed for PS@SiO2 motors when compared to their PS counterparts. We made use of stochastically optical reconstruction microscopy (STORM) to precisely detect single urease molecules conjugated to the micromotors surface with a high spatial resolution. An asymmetric distribution of enzymes around the micromotor surface was observed for both PS and PS@SiO2 architectures, indicating that the enzyme distribution was not the only parameter affecting the motion behavior. We quantified the number of enzymes present on the micromotor surface and observed a 10-fold increase in the number of urease molecules for PS@SiO2 motors compared to PS-based micromotors. To further investigate the number of enzymes required to generate a self-propulsion, PS@SiO2 particles were functionalized with varying amounts of urease molecules and the resulting speed and propulsive force were measured by optical tracking and optical tweezers, respectively. Surprisingly, both speed and force depended in a nonlinear fashion on the enzyme coverage. To break symmetry for active propulsion, we found that a certain threshold number of enzymes molecules per micromotor was necessary, indicating that activity may be due to a critical phenomenon. Taken together, these results provide new insights into the design features of micro/nanomotors to ensure an efficient development.