We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Enantioselective Fluorogenic Assay of Acetate Hydrolysis for Detecting Lipase Catalytic Antibodies.
- Authors
Klein, Gérard; Reymond, Jean-Louis
- Abstract
An enantioselective fluorogenic assay for the kinetic resolution of chiral alkyl acetates is demonstrated with 7-(3-acetoxybutoxy)-2 H-1-benzopyran-2-ones ( R)- and ( S)- 4 or 7-(3-acetoxy-2-methylpropoxy)-2 H-1-benzopyran-2-ones ( R)- 4 and ( S)- 6. The alcohols released by hydrolysis of these acetates are oxidized by horse-liver alcohol dehydrogenase to unstable β-(aryloxy)carbonyl compounds, which undergo β-elimination of the strongly fluorescent product umbelliferone (=7-hydroxy-2 H-1-benzopyran-2-one; 3) ( λem=460±20 nm, λex=360±20 nm). Enantioselectivities are calculated from the reaction rates for each enantiomeric acetate. For a series of representative lipases, the reactivities and enantioselectivities under preparative conditions are predicted accurately. This highly sensitive enantioselective assay detects as little as 10 μg/ml of hydrolytic enzyme, can be carried out in 96-well microtiter plates, and is compatible with cell-culture media. It is, therefore, suited for screening libraries of antibodies for enantioselective lipase catalytic antibodies.
- Publication
Helvetica Chimica Acta, 1999, Vol 82, Issue 3, p400
- ISSN
0018-019X
- Publication type
Article
- DOI
10.1002/(SICI)1522-2675(19990310)82:3<400::AID-HLCA400>3.0.CO;2-C