A Bioinspired “Plug-and-Play” Platform for the Multienzyme Cascade Synthesis of Purpurogallin

The precise organization of multienzymes on biomaterial surfaces remains a fundamental challenge in designing high-performance biohybrid systems. In this work, a bioinspired, hypothesis-driven platform, multitemplate artificial antibody-antigen directed immobilization (MT-3ADI), is introduced that enables programmable modular assembly of multiple enzymes. This "plug-and-play" architecture dictates structure-function relationships while facilitating enzyme exchange and system regeneration. Applied to purpurogallin synthesis, the engineered glucose oxidase (GOx)-horseradish peroxidase (HRP) and cellulase (CEL)-GOx-HRP cascades achieved exceptional yields (96.31 ± 0.13% and 90.25 ± 0.21%) and robust operational stability (retaining >80% activity after six cycles). By establishing a generalizable framework for controlled protein organization, this work addresses core priorities in biomaterial surface science and offers broad applicability from green synthesis to therapeutic delivery.