Abstract
Asymmetry in enzymatically driven nanomotor design, both structural and functional, is widely considered essential for propulsion. However, the interplay between particle geometry, enzyme distribution, and catalytic loading remains poorly defined, largely due to limited control over enzyme positioning that hinders quantitative analysis. Using DNA origami nanorods, we achieve precise spatial placement of urease enzymes with independently tunable coverage and asymmetry. Single-particle tracking reveals that motility arises not solely from enzyme number or spatial arrangement but from a balance between catalytic loading and geometric anisotropy. Unexpectedly, maximal propulsion occurs at ∼25% urease end-coverage, significantly below the conventional 50% end-coverage, where half of the available binding positions on one structural half of the origami are occupied. Boundary Element Method simulations incorporating identical spatial parameters reproduce these findings, confirming that programmable enzyme patterning dictates diffusiophoretic propulsion. These results provide a quantitative framework linking topology, catalytic activity, and motion, revealing that optimal motility does not coincide with maximal asymmetry and advancing the rational design of enzyme-powered DNA nanomotors.