The first fluorogenic assay for detecting a Baeyer-Villigerase activity in microbial cells.
The first fluorogenic assay allowing for detection of microbial enzymes able to perform Baeyer-Villiger oxidation is described. This is based on the use of 4-oxopentyl umbelliferyl ether 1 as a fluorogenic substrate. When Baeyer-Villigerases active against this test ketone are present in the selected whole cells, 1 is transformed into 3-hydroxypropyl umbelliferyl ether 3, which, in a subsequent step, releases the fluorescent product umbelliferone. Different microorganisms, known to be endowed with Baeyer-Villigerase activity, were assayed.
Gutiérrez
María C
MC
Groupe Biocatalyse et Chimie Fine UMR-CNRS 6111, Université de la Mediterranée, Faculté des Sciences de Luminy, case 901, 163 Avenue de Luminy, 13288 Marseille, Cedex 9, France.
Sleegers
Arthur
A
Simpson
Helen D
HD
Alphand
Véronique
V
Furstoss
Roland
R
D001419:Bacteria / Q000201:enzymology*; D001431:Bacteriological Techniques; D001681:Biological Assay; D004798:Enzymes / Q000032:analysis*; D005456:Fluorescent Dyes / Q000378:metabolism*; D006651:Histocytochemistry / Q000379:methods; D008956:Models, Chemical; D010084:Oxidation-Reduction
D004798:Enzymes; D005456:Fluorescent Dyes