Aflatoxin B1 (AFB1) is a well-established potent food mycotoxin. We previously evidenced that laccase could restrain AFB1’s genetic and oxidative toxicity in vitro conditions; nevertheless, there is no report about the in vivo toxicity of the laccase-mediated metabolites of aflatoxin B1 (LM-AFB1). In this investigation, we evaluated the in vivo safety of LM-AFB1 in comparison with the parent toxin using serum liver function tests (LFTs), oxidative stress biomarkers, histopathological examinations, and CYP1A2 and GSTA3 gene expression. Adult male mice were divided into four groups: (1) Control; (2) 4 mg/kg AFB1; (3) 4 mg/kg LM-AFB1; (4) 4 mg/kg intermediate transformational metabolites of AFB1 (CA-AFB1). In this study, AFB1’s liver toxicity was confirmed but LM-AFB1 did not significantly change the values. Gene expression analysis indicates that the utilization of laccase for bioremediation does not result in the suppression of glutathione S-transferase, in contrast to AFB1. Our research findings demonstrate that laccase has the potential to mitigate AFB1-induced liver toxicity in mice by averting oxidative stress caused by AFB1.