The development and application in different fields of multifunctional plasmonic nanoparticles (NPs) have always been research hotspots. Herein, multi-tip Au nanostars (NSs) with an anisotropic structure were fabricated for the photothermal therapy (PTT) of bacteria and surface-enhanced Raman scattering (SERS) detection of pollutants. The size and localized surface plasmon resonance (LSPR) characteristics of Au NSs were adjusted by varying Au seed additions. In addition, photothermal conversion performance of Au NSs with various Au seed additions was evaluated. Photothermal conversion efficiency of Au NSs with optimal Au seed additions (50 μL) was as high as 28.75% under 808 nm laser irradiation, and the heat generated was sufficient to kill (). Importantly, Au NSs also exhibited excellent SERS activity for the 4-mercaptobenzoic acid (4-MBA) probe molecule, and the local electromagnetic field distribution of Au NSs was explored through finite-difference time-domain (FDTD) simulation. As verified by experiments, Au NSs' SERS substrate could achieve a highly sensitive detection of a low concentration of potentially toxic pollutants such as methylene blue (MB) and bilirubin (BR). This work demonstrates a promising multifunctional nanoplatform with great potential for efficient photothermal inactivation and ultra-sensitive SERS detection.