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Aflatoxin B₁ (AFB₁) is a potent mycotoxin with mutagenic, carcinogenic, teratogenic, hepatotoxic, and immunosuppressive properties. In order to develop a bioremediation system for AFB₁-contaminated foods by white-rot fungi or ligninolytic enzymes, AFB₁ was treated with manganese peroxidase (MnP) from the white-rot fungus Phanerochaete sordida YK-624. AFB₁ was eliminated by MnP. The maximum elimination (86.0%) of AFB₁ was observed after 48 h in a reaction mixture containing 5 nkat of MnP. The addition of Tween 80 enhanced AFB₁ elimination. The elimination of AFB₁ by MnP considerably reduced its mutagenic activity in an umu test, and the treatment of AFB₁ by 20 nkat MnP reduced the mutagenic activity by 69.2%. ¹H-NMR and HR-ESI-MS analysis suggested that AFB₁ is first oxidized to AFB₁-8,9-epoxide by MnP and then hydrolyzed to AFB₁-8,9-dihydrodiol. This is the first report that MnP …