Laccases　are　a　class　of　multi-copper　oxidases　with　important　industrial　values.　A　thermotolerant　laccase　produced　by　a　basidiomycete　fungal　strain　Cerrena　unicolor　CGMCC　5.1011　was　studied.　With　glycerin　and　peptone　as　the　carbon　and　nitrogen　sources,　respectively,　a　maximal　laccase　activity　of　121.7　U/mL　was　attained　after　cultivation　in　the　shaking　flask　for　15 days.　Transcriptomics　analysis　revealed　an　expressed　laccase　gene　family　of　12　members　in　C.　unicolor　strain　CGMCC　5.1011,　and　the　gene　and　cDNA　sequences　were　cloned.　A　glycosylated　laccase　was　purified　from　the　fermentation　broth　of　Cerrena　unicolor　CGMCC　5.1011　and　corresponded　to　Lac2　based　on　MALDI-TOF　MS/MS　identification.　Lac2　was　stable　at　pH　5.0　and　above,　and　was　resistant　to　organic　solvents.　Lac2　displayed　remarkable　thermostability,　with　half-life　time　of　1.67 h　at　70 ºC.　Consistently,　Lac2　was　able　to　completely　decolorize　malachite　green　(MG)　at　high　temperatures,　whereas　Lac7　from　Cerrena　sp.　HYB07　resulted　in　accumulation　of　colored　MG　transformation　intermediates.　Molecular　dynamics　simulation　of　Lac2　was　conducted,　and　possible　mechanisms　underlying　Lac2　thermostability　were　discussed.　The　robustness　of　C.　unicolor　CGMCC　5.1011　laccase　would　not　only　be　useful　for　industrial　applications,　but　also　provide　a　template　for　future　work　to　develop　thermostable　laccases.　©　2020,　The　Author(s).