title: Fluorogenic substrates for lipases, esterases, and acylases using a TIM-mechanism for signal release.
abstract: 3-Acyloxyl-2-oxopropyl ethers of umbelliferone were investigated as new fluorogenic substrates for lipases and esterases. The aliphatic primary alcohol-leaving group released the fluorescent product umbelliferone by an enolization/beta-elimination reaction similar to the triose phosphate isomerase (TIM) reaction. A similarly designed phenylacetamide provided a fluorescent probe for penicillin G acylase, whereby the enolization/beta-elimination sequence from the intermediate aminoketone was very fast and spontaneous even under acidic conditions. The corresponding epoxyketone was not fluorogenic with epoxide hydrolases (EH). These substrates represent periodate-free Clips-otrade mark substrates.
authors.affiliation: Department of Chemistry and Biochemistry, University of Berne, Berne, Switzerland.
authors.forename: Renaud
authors.initials: R
authors.lastname: Sicart
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Collin
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Reymond
mesh_terms: D000081:Acetamides / Q000737:chemistry / Q000378:metabolism; D004851:Epoxide Hydrolases / Q000378:metabolism; D004950:Esterases / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism*; D006868:Hydrolysis; D008049:Lipase / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D015394:Molecular Structure; D010399:Penicillin Amidase / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D014468:Umbelliferones / Q000737:chemistry / Q000378:metabolism
chemical_list: D000081:Acetamides; D005456:Fluorescent Dyes; D014468:Umbelliferones; C031477:7-hydroxycoumarin; C030686:acetamide; D004950:Esterases; D008049:Lipase; D004851:Epoxide Hydrolases; D010399:Penicillin Amidase