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Hydrolysis of 4‐methylumbelliferyl butyrate: A convenient and sensitive fluorescent assay for lipase activity
Author(s) -
Roberts Ingram M.
Publication year - 1985
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02534195
Subject(s) - fluorometer , lipase , chemistry , fluorescence , chromatography , hydrolysis , substrate (aquarium) , butyrate , lecithin , biochemistry , micelle , nile red , enzyme , organic chemistry , aqueous solution , biology , fermentation , physics , quantum mechanics , ecology
A rapid, sensitive and convenient fluorescent assay was developed to screen for lipase activity. The non‐fluorescent substrate, 4‐methylumbelliferyl butyrate (4‐MUB), solubilized in either liposomal dispersions or bile salt/lecithin mixed micelles, is hydrolyzed to butyric acid and the highly fluorescent compound, 4‐methylumbelliferone (4‐MU). Assays are run at 37 C for 10 min, terminated, and the changes in fluorescence quantitated with a Turner III fluorometer. Both lingual and pancreatic lipases exhibit activity against this artificial substrate. The assay has several advantages: nmoles 4 MU/ml/hr are measured allowing the detection of very low lipolytic activity; multiple samples may be simultaneously assayed, and only a brief incubation period is required.
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