Low background FRET-substrates for lipases and esterases suitable for high-throughput screening under basic (pH 11) conditions.
FRET-based fluorogenic substrates for lipases and esterases were prepared in four steps from commercially available building blocks. The substrates are pyrenebutyric acid monoesters of aliphatic 1,2-diols bearing a dinitrophenylamino group as a quencher. The most enzyme-reactive substrate is ester 2a. The substrates do not show any measurable background reaction in the absence of enzyme even at pH 11, but react fast and specifically with lipases and esterases. These substrates offer an unprecedented and practical solution to the long-standing problem of a simple yet efficient high-throughput screening tool for lipase activities under basic conditions.
Yang
Yongzheng
Y
Department of Chemistry & Biochemistry, University of Berne, Switzerland.
Babiak
Peter
P
Reymond
Jean-Louis
JL
D004136:Dinitrobenzenes; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004950:Esterases / Q000187:drug effects*; D004952:Esters / Q000493:pharmacokinetics; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000187:drug effects*; D011721:Pyrenes; D013379:Substrate Specificity
D004136:Dinitrobenzenes; D004952:Esters; D005456:Fluorescent Dyes; D011721:Pyrenes; C002218:1-pyrenebutyrate; D004950:Esterases; D008049:Lipase