Pyrenemethyl laurate, a new fluorescent substrate for continuous kinetic determination of lipase activity.
A fluorescent acyl derivative of pyrenemethanol, pyrenemethyl laurate, was synthesized and used for the determination of several lipases by a continuous kinetic assay. The influence of the physical parameters of the substrate (pyrenemethyl laurate) and its hydrolysis product (pyrenemethanol), on the fluorescence emission was studied. The hydrolysis of pyrenemethyl laurate could be monitored directly in a spectrofluorometer because of the very high monomeric emission of pyrenemethanol at about 375 nm, whereas an aqueous dispersion of pyrenemethyl laurate emitted at 475 nm ('excimeric'). Pyrenemethyl laurate was hydrolyzed by gastric lipase, cellular lipases of haemopoietic cells, and the bacterial lipase of Rhizopus arrhizus.
Nègre
A
A
Laboratoire de Biochimie, Faculté de Médecine, Toulouse, France.
Salvayre
R
R
Dagan
A
A
Gatt
S
S
D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D007848:Laurates; D007850:Lauric Acids; D008049:Lipase / Q000378:metabolism*; D011721:Pyrenes; D012233:Rhizopus / Q000201:enzymology; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D005456:Fluorescent Dyes; D007848:Laurates; D007850:Lauric Acids; D011721:Pyrenes; C061646:pyrenemethyl laurate; D008049:Lipase