We experimentally investigated the bi-anisotropic properties of Fano resonance in three-dimensional (3D) metamaterials. Fano resonance in 3D metamaterials arises from the interference of in-phase and anti-phase modes that originate from mode hybridization in coupled 3D split ring resonators (SRRs) with detuned resonant wavelengths. At Fano resonance, not only permittivity and permeability but also the bi-anisotropic parameter show doubly dispersive response. Manipulation of the bi-anisotropic response at Fano resonance was demonstrated through controlling the inversion symmetry of the 3D-SRRs. Improvement of inversion symmetry due to rotation of 3D-SRRs results in enhancement of magnetic response and inhibition of electric and bi-anisotropy responses at Fano resonance. Negligible electric and bi-anisotropic responses at Fano resonance were achieved due to the small radiative nature of the anti-phase mode. This bi-anisotropic Fano metamaterials with rich and tunable bi-anisotropy will extend the capabilities of new optical phenomena and broaden the applications of bi-anisotropic metamaterials.