A Versatile Periodate-Coupled Fluorogenic Assay for Hydrolytic Enzymes

Abstract

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 (λex=360 nm). This simple assay is suitable for high-throughput screening.