Synthesis of metal oxide nanoparticles using novel methodologies always attracts great importance in research. The use of plant extract to synthesize nano-particle has been considered as one of the eco-friendly methods. This paper describes the biosynthetic route of preparation of zinc oxide nanoparticles (ZnO NPs) from the Lagerstroemia speciosa leaf extract. This approach appears to be low-cost preparation and alternative method to conventional methods. Highly stable and hexagonal phase ZnO NPs with average particle size of 40nm were synthesized and characterized by UV-Vis absorption spectroscopy (surface Plasmon resonance), Fourier transform infrared spectroscopy (surface functionalities), X-ray Diffraction analysis (crystallinity), TEM and SEM (size and morphology), Energy Dispersive X-ray spectroscopy (elemental composition), Thermogravimetric analysis (weight loss) and Zeta potential (stability). The preliminary phytochemical experiments identify the possible chemical groups present in leaves extract. The photocatalytic properties of ZnO NPs were studied using UV-Vis spectroscopy by exposing methyl orange to sunlight and it is found to be degraded up to 93.5% within 2h. The COD values were significantly reduced from 5600mg/L to 374mg/L after 100min of solar radiation. The hemolytic activity of synthesized zinc oxide nanoparticles was performed on human erythrocyte cells. Thus the present study provides a simple and eco-friendly method for the preparation of multifunctional property of ZnO NPs utilizing the biosynthetic route.