Synergistic Co-Assembly of Natural Oxidase and Peptide-Heme Nanozyme for In Situ H₂O₂ Removal with Enhanced Catalytic Efficiency and Stability.

Abstract

Hydrogen peroxide, a byproduct of the oxidase catalytic process, significantly compromises the enzymatic activity and stability of natural oxidases, thereby restricting their industrial applications. To address this challenge, we developed a coassembly system combining a natural oxidase with a peptide-heme (PepHeme) nanozyme possessing catalase-like activity on a ferritin platform. This system, demonstrated by using natural choline oxidase and galactose oxidase as models, forms an oxidase-catalase cascade reaction. Leveraging the self-assembly properties of ferritin and the directional association enabled by sortase ligation, we successfully constructed clusters of oxidases and PepHeme nanozymes. This design facilitated the directional immobilization of natural enzymes and nanozymes and increased the local concentration of these cascade clusters, thus significantly enhancing both the catalytic efficiency and stability of the enzyme, especially the hydrogen peroxide tolerance and long-term stability. This cascade system offers a promising solution to the hydrogen peroxide byproduct issue in the industrial applications of oxidases.