A Versatile Periodate-Coupled Fluorogenic Assay for Hydrolytic Enzymes

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.