In this work, we described a green method for creating TiO2 nanoparticles by employing Wisteria floribunda (WF) as a capping and reducing agent. SEM, FTIR, UV–VIS spectroscopy, morphology, particle size, energy dispersive spectroscopy (EDS), elemental composition, crystalline structure, X-ray diffraction (XRD), and stability were among the many distinctive factors that were investigated. In the instance of UV–VIS spectroscopy, a distinctive TiO2 peak was seen at 340 nm, confirming the formation of TiO2 nanoparticles. Subsequent morphological analyses revealed that the shape was spherical and that the elemental composition of titanium was 32.07%, whereas that of oxygen was 31.06%. TiO2 particles showed distinct stretching and bending peaks in the FTIR. Antimicrobial activity was then conducted alongside a kind of gram-positive microorganisms and gram-negative microorganisms.