Enzyme-Instructed Interfacial Jamming of Pillar[5]arenes for Macroscopic Signal Amplification.

Enzyme-instructed signal generation at liquid-liquid interfaces presents a novel strategy for controlling and detecting biochemical processes on macroscopic scales. Here, we explore the self-assembly and jamming of pillar[5]arene (P[5]A) derivatives at the oil-water interface via a copper-mediated "click" reaction, providing a versatile platform for generating observable signals. The formation of a pillar[5]arenes network at the droplet interface reduces interfacial tension, allowing droplets to adopt various nonequilibrium shapes based on the interfacial jamming process. By varying concentrations of P[5]A derivatives and ascorbic acid (AA), we fine-tune the surface coverage of droplets, offering control over the jamming dynamics. Additionally, we introduce a signal amplification mechanism where the dephosphorylation of a dormant reductant by alkaline phosphatase (ALP) triggers the "click" reaction at the interface. This system enables the quantification of ALP activity through macroscopic surface changes with inhibition of ALP by heavy metals and metal chelators reducing surface coverage. This approach represents a promising method for amplifying molecular signals into detectable macroscopic outputs with potential applications in biochemical sensing and materials science.