Determination of Lipase Activity in Organic Solvent Using Fluorescent Substrate and Its Application to the Screening of Lipase Inhibitor

Measurement of the activity of a lipase-detergent complex in a homogeneous system using 4-methylumbelliferyl oleate as a fluorescent substrate was studied. A linear relation between the quantity of the complex and the amounts of 4-methylumbelliferone formed was obtained up to 240pmol/tube when incub...

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Bibliographic Details
Published in:Analytical Sciences 1991/02/10, Vol.7(1), pp.15-18
Main Authors: SHIMURA, Susumu, TSUZUKI, Wakako, SUZUKI, Tateo
Format: Article
Language:English
Subjects:
4-methylumbelliferyl oleate
Analytical, structural and metabolic biochemistry
Biological and medical sciences
enzyme inhibitor
Enzymes and enzyme inhibitors
fluorometry
Fundamental and applied biological sciences. Psychology
Lipase
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Summary:Measurement of the activity of a lipase-detergent complex in a homogeneous system using 4-methylumbelliferyl oleate as a fluorescent substrate was studied. A linear relation between the quantity of the complex and the amounts of 4-methylumbelliferone formed was obtained up to 240pmol/tube when incubated with 0.05mM 4-methylumbelliferyl oleate in aqueous 95% tetrahydrofuran. Under these conditions, the activity of the complex is about 1pmol min-1 μg-1. The maximum activity was obtained in neutral or weakly alkaline medium. In this assay system, metal ions such as Fe3+ and Co2+ strongly inhibited the activity. This method could be applied to the screening of lipase inhibitors in foods.
ISSN:0910-6340
1348-2246
DOI:10.2116/analsci.7.15