Programmable biofunctionalization of bacterial cellulose via a dynamic co-culture platform.

Abstract

Programmable biofunctionalization of bacterial cellulose (BC) is promising for constructing engineered living materials, but current microbial co-culture approaches struggle to combine coherent matrix formation, dynamic cultivation, and efficient matrix-associated protein functionalization. In this study, we report a polydopamine (PDA)-mediated, surface-immobilized dynamic co-culture platform that pairs Komagataeibacter rhaeticus for BC production with engineered Pichia pastoris for recombinant protein secretion. PDA-mediated co-immobilization preserves conformal BC growth under agitation while maintaining the functional partner near the forming matrix. Cellulose-binding-domain fusion further retains secreted protein cargoes within the BC network, enabling localized functionalization. The template-guided strategy supports geometry control, construct-size scaling, and incorporation of multiple engineered yeast populations. Modular genetic payloads enable pollutant degradation and enzymatic cascade-based analyte detection. As an application-relevant demonstration, lysostaphin-functionalized BC hydrogels effectively controlled Staphylococcus aureus infection and accelerated wound closure in a diabetic murine model. This work establishes a functionally extensible framework for bioactive BC-based living materials.