Simultaneous measurements of enzyme propagation and biopolymer mobility elucidate the directional degradation of a dense biopolymer matrix

Abstract

Enzymatic degradation of biopolymers underpins critical processes in biotechnological applications and natural processes, such as food digestion and cancer development, yet the interplay between enzyme propagation in a dense substrate and substrate degradation remains unresolved. It remains especially unclear whether degradation of a dense matrix facilitates or impedes enzyme propagation. We simultaneously track with spatiotemporal resolution unidirectional enzyme diffusion and biopolymer degradation in a model system. We demonstrate that enzyme diffusion is decoupled from catalytic activity, while the degradation front progression is dictated by enzyme diffusion, reaction kinetics and slow enzyme deactivation. These findings establish a quantitative framework to optimize enzymatic processes for applications in biomedicine, biomass valorization, and nanotechnology.