Enantioselective Fluorogenic Assay of Acetate Hydrolysis for Detecting Lipase Catalytic Antibodies

Studies on whole cell fluorescence-based screening for epoxide hydrolases and Baeyer-Villiger monoxygenases

“…Epoxides, probe-substrates for EH, were known to endure the assay conditions. 4,12 The same was not true for ketone 9, therefore its stability in borate buffer pH 8.8 had to be evaluated in the presence and in the absence of the assay components (Scheme 2), namely horse liver alcohol dehydrogenase (HLADH), NAD + and BSA. No time-dependent increase in fluorescence was observed for any combination of reagents within 48 h, discarding the formation of 11 and release of 7 in the absence of a specific Baeyer-Villiger monooxygenase.…”

“…Although high throughput screening (HTS) assays are being rapidly innovated, 2,9-11 whole cell HTS assays have been almost neglected, excepting those based on fluorogenic substrates which have been applied to metagenomic libraries (in function-driven analysis) but seldom to natural culture collections. In order to address this problem, this paper reports the application of a 96-well microplate fluorogenic assay, originally designed for purified hydrolytic enzymes, 12,13 to screen microbial whole cells for epoxide hydrolases (EHs) and Baeyer-Villiger monooxygenases (BVMOs).…”

Fluorogenic substrates for the screening assay of transketolase through beta-elimination of umbelliferone—Development, scope and limitations

“…Following this general strategy, new assays were described for lipases and esterases (Klein and Reymond, 1999;Badalassi et al, 2000;Nyfeler et al, 2003;Leroy et al, 2003) (including epoxide hydrolases EH, Badalassi et al, 2000), acylases (Badalassi et al, 2000), phosphatases (Badalassi et al, 2000;Gonzalez-Garcia et al, 2003a), proteases (Badalassi et al, 2002) and Baeyer-Villigerase (Gutiérrez et al, 2003;Sicard et al, 2005).…”

Synthesis of a new long-wavelength latent fluorimetric indicator for analytes determination in the DT-Diaphorase coupling dehydrogenase assay system

“…The chemical structural of BQRh. fluorescent coumarin or rhodamine derivatives upon activation by enzymes had been developed as sensitive tools for monitoring specific enzymatic reactions in diagnostic applications (Klein and Reymond, 1999;Kupcho et al, 2004;Zlokarnik et al, 1998;Huang and Lin, 2006).…”