Flow-driven structural and transcriptomic responses in syntrophic electroactive biofilms

Bioelectrochemical systems (BES) are versatile platforms for energy and resource recovery, where electroactive biofilms serve as the key biocatalyst. While performance depends on flow dynamics, the interplay between hydrodynamic conditions, biofilm architecture, and gene-regulation in syntrophic communities remains poorly understood. We investigated defined surface flow velocities (0.2 and 0.8 mm/s) on the structure and electroactivity of Shewanella oneidensis-Geobacter sulfurreducens coculture biofilms. Higher flow produced more compact biofilms with lower biovolume and porosity, yielding higher current densities. Flow transitions induced persistent structural and electrochemical shifts, indicating an adaptive, gene-regulated response to shear. Since G. sulfurreducens monoculture biofilms were flow-insensitive, regulation appears to originate from S. oneidensis. Transcriptomics showed strong shear response, including 10.7-fold upregulation of bpfD. Its deletion reduced current density and increased porosity despite comparable biovolume. Three-dimensional fluorescence in situ hybridization imaging revealed a porous, loosely organized structure within the mutant biofilm. This was consistent with the structural and electrochemical differences observed. These findings reveal that community-level electrochemical performance can be governed by a minor transcriptional contributor rather than by the dominant current-producing species.