Tyrosinase (TYR) which can catalyze the oxidation of catechol is recognized as a significant biomarker of melanocytic lesions, thus developing powerful methods for the determination of TYR activity is highly desirable for the early diagnosis of melanin-related diseases, including melanoma. Herein, we develop a novel portable and recyclable surface-enhanced Raman scattering (SERS) sensor, prepared by assembling gold nanoparticles and p-thiol catechol ( p-TC) on an ITO electrode, for detecting TYR activity via the SERS spectral variation caused by the conversion of p-TC into its corresponding quinone under TYR catalysis. The developed SERS sensor has a rapid response to TYR within 1 min under the optimized conditions and shows high selectivity for TYR with the detection limit at 0.07 U/mL. Importantly, this SERS sensor can be easily regulated by applying negative voltage to achieve circular utilization, favoring the automation of SERS detection. Furthermore, the presented recyclable SERS sensor can perform well on both the determination of TYR activity in serum and the assessment of TYR inhibitor, demonstrating huge potential in the sensitive, selective, and facile detection of TYR activity for disease diagnosis and drug screening related with TYR.