In the current study, we have reported a novel approach for synthesizing zinc oxide Nanoparticles (ZnO Nps) from (−)-Hydroxy Citric Acid [(−)-HCA] extracted from the dried rinds of the fruit Garcinia gummi-gutta. The prepared nanoparticles were characterized by UV-Vis spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Scanning electron microscopy (SEM), Energy dispersive X-ray analysis (EDAX) and High-resolution transmission electron microscopy (HR-TEM). Hydroxy citric acid (HCA) was used as a template to synthesize the ZnO Nps. The presence of (−)-HCA was confirmed by High-performance liquid chromatography (HPLC). Spherical clusters built of nanoparticle aggregates with the desired composition of ZnO were confirmed from the SEM-EDAX and HRTEM analysis. The synthesized ZnO Nps showed a positive response for antibacterial, antifungal, antioxidant and cytotoxicity activities. © 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim