A new strategy about the biological way to modulate intrinsic carbon defects of the biochar framework was developed. Herein, the porous carbon/Fe 3 O 4 /Ag (PC/Fe 3 O 4 /Ag) composites are first prepared using a novel fungi-enabled method, and the mechanism underlying their excellent catalytic and adsorption properties was elucidated. Customization of microstructures and introducing defects in the carbon-based nanomaterial can be achieved through regulation of the fungus growth process on biomass, thereby improving the catalytic performance greatly. This work also opened a new area in materials science based on fungi-enabled defect engineering and pore channel regulation, providing unique insights into environment treatment. Theoretical calculations and simulations were carried out to understand the remarkable catalytic and adsorption performance.