Aflatoxin　B1　(AFB1)　is　a　known　toxic　human　carcinogen　and　can　be　detoxified　by　laccases,　which　are　multicopper　oxidases　that　convert　several　environmental　pollutants　and　toxins.　In　this　study,　a　new　laccase　that　could　catalyze　AFB1　degradation　was　purified　and　identified　from　the　white-rot　fungus　Cerrena　unicolor　6884.　The　laccase　was　purified　using　(NH4)2SO4　precipitation　and　anion　exchange　chromatography,　and　then　identified　as　Lac　2　through　zymogram　and　UHPLC-MS/MS　based　on　the　Illumina　transcriptome　analysis　of　C.　unicolor　6884.　Six　putative　laccase　protein　sequences　were　obtained　via　functional　annotation.　The　lac　2　cDNA　encoding　a　full-length　protein　of　512　amino　acids　was　cloned　and　sequenced　to　expand　the　fungus　laccase　gene　library　for　AFB1　detoxification.　AFB1　degradation　by　Lac　2　was　conducted　in　vitro　at　pH　7.0　and　45　◦C　for　24　h.　The　half-life　of　AFB1　degradation　catalyzed　by　Lac　2　was　5.16　h.　Acetosyringone　(AS),　Syrinagaldehyde　(SA)　and　[2,20　-azino-bis-(3-ethylbenzothiazoline-6-sulfonic　acid)]　(ABTS)　at　1　mM　concentration　seemed　to　be　similar　mediators　for　strongly　enhancing　AFB1　degradation　by　Lac　2.　The　product　of　AFB1　degradation　catalyzed　by　Lac　2　was　traced　and　identified　to　be　Aflatoxin　Q1　(AFQ1)　based　on　mass　spectrometry　data.　These　findings　are　promising　for　a　possible　application　of　Lac　2　as　a　new　aflatoxin　oxidase　in　degrading　AFB1　present　in　food　and　feeds.　©　2020　by　the　authors.　Licensee　MDPI,　Basel,　Switzerland.　This　article　is　an　open　access　article　distributed　under　the　terms　and　conditions　of　the　Creative　Commons　Attribution　(CC　BY)　license　(http://creativecommons.org/licenses/by/4.0/).