Tetrahydrofuran-3-one　(3TF)　was　regarded　as　a　difficult-to-reduce　ketone　for　carbonyl　reductases　due　to　its　high　stereosymmetry,　result　of　which,　the　biosynthesis　of　chiral　3-hydroxytetrahydrofuran　(3HTF),　a　key　precursor　of　pharmaceuticals　for　treatment　of　HIV　or　diabetes,　has　been　limited.　Present　study　mined　a　robust　carbonyl　reductase　CmCR　from　Candida　metapsilosis　with　high　activity　towards　3TF　using　isopropanol　as　co-substrate　(500　mM,　22　g/L/h　space-time　yield,　67%　ee).　Based　on　the　orientation　of　substrate＇s　etheryl　oxygen　in　the　substrate　binding　pocket,　CmCR　was　rationally　designed　and　two　mutants　MuR　and　MuS　were　screened　out　to　completely　reduce　3TF　into　(R)-HTF　and　(S)-HTF　respectively　with　the　highest　3TF　loading　and　productivity　(200　mM,　4.4　g/　L/h　space-time　yield,　99%　ee　for　MuR　while　500　mM,　11　g/L/h　space-time　yield,　99%　ee　for　MuS).　The　structural　mechanism　for　the　enhanced　stereoselectivity　was　revealed　that　the　mutagenesis　changed　the　electrostatic　potential　surrounding　the　substrate　entrance　and　promoted　3TF　to　approach,　bind　and　form　prereaction　state　with　MuR　or　MuS　in　one　certain　direction,　by　which　3TF　was　converted　into　3HTF　with　excellent　optical　purity.　The　success　in　this　study　provides　a　viable　approach　for　rational　design　of　carbonyl　reductases　with　high　enantioselectivity　towards　target　substrates　of　high　symmetry.