A low background high-Throughput screening (HTS) fluorescence assay for lipases and esterases using acyloxymethylethers of umbelliferone

Esters and acyloxymethyl ethers of umbelliferone were evaluated as fluorogenic substrates for lipases and esterases with respect to stability and resistance to non-specific hydrolysis. Isobutyryloxymethyl ether 1c and pivaloxymethyl ether 1d were found to be optimal substrates for enzyme assays, particularly with respect to HTS applications.