Universal chromogenic substrates for lipases and esterases

Fluorescence Assays for Biotransformations

“…An enzyme fi ngerprinting study involving eight enantiomeric pairs of analogs of 19 with various acyl chain length showed that lipases and esterases can be distinguished by their chain length selectivity [34] . The corresponding nitrophenyl and dinitrophenyl derivatives 22 and 23 display similar reactivities [35] . As for the ADH and aldolase assays discussed above (Scheme 1.1 ), the secondary decomposition of the 1,2 -diol product leading to umbelliferone may be kinetically limiting in the assay, implying that the substrates are not suitable for following the kinetics of very fast enzymes.…”

Alteration of lipase properties by protein engineering methods

“…Furthermore, the substrates are often unstable at extreme pH or temperature and this autohydrolysis can limit their use in certain screening efforts. Alternative compounds were suggested by Reymond et al coupled with a periodate treatment and b-elimination to release the chromophore/fluorophore [30][31][32]. However, these substrates have to be chemically synthesized and only end-point measurements are possible.…”

Multienzyme Profiling of Thermophilic Microorganisms with a Substrate Cocktail Assay

“…Multisubstrate assays are called activity profiling or fingerprinting. [5] Activity fingerprints of single enzymes using structurally diverse substrates allow a functional classification of closely related enzymes, as has been demonstrated, for example, for lipases, [6] proteases, [7] or alcohol dehydrogenases. [8] At the level of microbial cultures, multisubstrate assays can be carried out addressing different enzyme activities.…”