BACKGROUND: Cancer is one of the leading causes of death worldwide, highlighting the need to develop novel therapeutic strategies that are more effective and have fewer side effects than conventional treatments. Enzymatic cancer therapy is a promising approach due to its high specificity and minimal toxicity. Among the various enzymes, laccase, a widely used biocatalyst, has shown significant potential for anti-cancer applications due to its proliferation inhibitory properties. METHODS AND RESULTS: In this study, the enzyme laccase from Trametes versicolor was purified by three-phase partitioning and then its cytotoxic, genotoxic and apoptotic effects on thyroid cancer (TT) and endometrial cancer (Ishikawa) cell lines were investigated. Laccase exhibited IC CONCLUSIONS: Our results emphasize the specific cytotoxicity and molecular mechanisms underlying the anti-cancer effect of laccase and demonstrate that laccase is capable of selectively targeting cancer cells and causing apoptosis and DNA damage. This study demonstrates the potential of laccase as a novel enzymatic therapeutic for the treatment of thyroid and endometrial cancer and warrants further investigation into its clinical application and efficacy.