An enantioselective fluorimetric assay for alcohol dehydrogenases using albumin-catalyzed β-elimination of umbelliferone

3-hydroxybutyl umbelliferyl ethers (R)-1 and (S)-1 are fluorogenic substrates for alcohol dehydrogenases. Their oxidation forms ketone 2, which undergoes β-elimination of umbelliferone under catalysis by bovine serum albumin, leading to a> 20-fold fluorescence increase at λem = 460 ± 20 nm (λex = 360 ± 20 nm). Enantioselectivity is determined in two separate tests with each enantiomeric substrate