Nanomaterials‐based artificial enzymes (AEs) have flourished for more than a decade. However, it is still challenging to further enhance their biocatalytic performances due to the limited strategies to tune the electronic structures of active centers. Here, a new path is reported for the de novo design of the d electrons of active centers by modulating the electron transfer in vanadium‐based AEs (VOx‐AE) via a unique Zn–O–V bridge for efficient reactive oxygen species (ROS)‐catalysis. Benefiting from the electron transfer from Zn to V, the V site in VOx‐AE exhibits a lower valence state than that in V2O5, which results in charge‐filled V‐dyz orbital near the Fermi level to interfere with the formation of sigma bonds between the V‐ dz2 and O‐pz orbitals in H2O2. The VOx‐AE exhibits a twofold Vmax and threefold turnover number than V2O5 when catalyzing H2O2. Meanwhile, the VOx‐AE shows enhanced catalytic eradication of drug‐resistant bacteria and achieves comparable wound‐treatment indexes to vancomycin. This modulating charge‐filling of d electrons provides a new direction for the de novo design of nanomaterials‐based AEs and deepens the understanding of ROS‐catalysis.