An enantioselective fluorimetric assay for alcohol dehydrogenases using albumin-catalyzed beta-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 beta-elimination of umbelliferone under catalysis by bovine serum albumin, leading to a > 20-fold fluorescence increase at lambda em = 460 +/- 20 nm (lambda ex = 360 +/- 20 nm). Enantioselectivity is determined in two separate tests with each enantiomeric substrate.
Klein
G
G
Department of Chemistry & Biochemistry, University of Bern, Switzerland.
Reymond
J L
JL
D000426:Alcohol Dehydrogenase / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D002417:Cattle; D005456:Fluorescent Dyes; D006736:Horses; D007202:Indicators and Reagents; D007700:Kinetics; D007778:Lactobacillus / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013237:Stereoisomerism; D013379:Substrate Specificity; D014468:Umbelliferones / Q000378:metabolism
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D014468:Umbelliferones; D012710:Serum Albumin, Bovine; D000426:Alcohol Dehydrogenase