Ethyl　3-hydroxy-3-phenylpropionate　(EHPP),　(R)-EHPP　or　(S)-EHPP,　is　an　important　chiral　intermediate　for　pharmaceuticals.　Its　synthesis　from　ethyl　benzoyl　acetate　(EBA)　by　alcohol　dehydrogenase　is　regarded　as　a　green　method.　However,　scarcely　any　alcohol　dehydrogenase　has　been　reported　competent　in　asymmetric　synthesis　of　chiral　EHPP　at　high　EBA　loading.　Present　study　developed　two　robust　and　efficient　bio-catalysts　Mu-S2　and　Mu-R4　for　preparation　of　(S)-EHPP　and　(R)-EHPP　respectively　by　rational　design　of　alcohol　dehydrogenase　PcSDR　from　Pedobacter　chitinilyticus　based　on　molecular　dynamics　(MD)　simulation　analysis.　BtGDH,　a　glucose　dehydrogenase　from　Bacillus　toyonensis　catalyzing　the　oxidation　of　glucose　for　cofactor　regeneration,　was　co-expressed　with　the　screened　mutants　to　form　enzyme　systems　Mu-S2-BtGDH　and　Mu-R4-BtGDH.　After　reaction　condition　optimization,　Mu-S2-BtGDH　and　Mu-R4-BtGDH　were　efficient　in　the　synthesis　of　(S)-EHPP　(94%　conv.　and　99%　e.e.)　and　(R)-EHPP　(99%　conv.　and　98%　e.e.)　respectively　in　100　mL　scale　under　500　mM　of　EBA　loading　in　10　h　following　a　substrate　continuous　feeding　mode.　After　purifying,　the　isolated　yield　for　each　EHPP　enantiomer　is　＞　93%.　This　work　not　only　provides　potential　biocatalysts　for　the　industrial　production　of　(R)-EHPP　and　(S)-EHPP,　but　also　enriches　the　constructure-function　relationship　of　alcohol　dehydrogenases.　©　2022　Elsevier　Inc.