In　this　paper,　horseradish　peroxidase　(HRP)　was　successfully　immobilized　on　heated　Au　disk　electrode　(HAuDE)　by　biotin-streptavidin　specific　interaction　through　HS-ssDNA-biotin　self-assembled　on　HAuDE　for　investigation　the　electrocatalytic　activity　of　HRP.　With　elevated　electrode　temperature,　the　significant　temperature　effect　of　the　electrocatalytic　activity　of　HRP　for　H2O2　reduction　was　demonstrated　by　using　this　bio-sensing　platform.　With　an　electrode　temperature　of　40　°C,　a　detection　limit　of　1.5　×　10−6　mol　L−1　for　H2O2　reduction　could　be　obtained,　which　was　more　than　one　magnitude　lower　than　that　with　an　electrode　temperature　of　0　°C.　Because　HRP　can　be　widely　used　as　an　enzyme　label　for　amplification　detection,　this　sensing　platform　can　be　broadly　applied　to　analytical　chemistry　such　as　nucleic　acid　detection,　and　aptamer-based　biosensors.　©　2019　Elsevier　B.V.