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A Versatile Periodate‐Coupled Fluorogenic Assay for Hydrolytic Enzymes
Author(s) -
Badalassi Fabrizio,
Wahler Denis,
Klein Gérard,
Crotti Paolo,
Reymond JeanLouis
Publication year - 2000
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/1521-3773(20001117)39:22<4067::aid-anie4067>3.0.co;2-9
Subject(s) - sodium periodate , umbelliferone , chemistry , periodate , bovine serum albumin , enzyme , hydrolysis , fluorescence , biochemistry , chromatography , aldehyde , coumarin , organic chemistry , catalysis , physics , quantum mechanics
Enzymes such as acylases , lipases, epoxide hydrolases, or phosphatases can be easily detected by fluorescence when they hydrolyze suitable precursors to produce the 1,2‐diols 1 and 1,2‐aminoalcohol 2 as products. Indeed, oxidation of 1 and 2 with sodium periodate and β ‐elimination of the intermediate aldehyde with bovine serum albumin yields umbelliferone, and results in a more than 20‐fold increase in fluorescence at 460 nm ( λ ex =360 nm). This simple assay is suitable for high‐throughput screening.