Determination of Lipase Activity by a Rhodamine-Triglyceride-Agarose Assay

A quantitative fluorescence lipase assay based on the interaction of rhodamine B with fatty acids released during the enzymatic hydrolysis of triglycerides is described. The assay is linear over the range of 0.5-2 mM oleic acid and 0.05-1 μg pure lipase. The method allows flexibility in the choice o...

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Published in:Analytical biochemistry 1994-06, Vol.219 (2), p.256-260
Main Authors: Jette, J.F., Ziomek, E.
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, Ion Exchange - methods
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Geotrichum - enzymology
Geotrichum - growth & development
Hydrolases
Indicators and Reagents
Kinetics
Lipase - analysis
Lipase - isolation & purification
Lipase - metabolism
Oleic Acid
Oleic Acids
Rhodamines
Sepharose
Spectrometry, Fluorescence - methods
Substrate Specificity
Triglycerides - metabolism
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creator Jette, J.F.
Ziomek, E.
description A quantitative fluorescence lipase assay based on the interaction of rhodamine B with fatty acids released during the enzymatic hydrolysis of triglycerides is described. The assay is linear over the range of 0.5-2 mM oleic acid and 0.05-1 μg pure lipase. The method allows flexibility in the choice of substrate. A large number of samples can be assayed simultaneously, making it practical for assaying lipase activity in column fractions during purification. The substrate profiles of Geotrichum candidum lipase obtained by both a titrimetric assay and the RTA assay indicated the highest activity against triolein. The method is rapid and can be further automated.
doi_str_mv 10.1006/abio.1994.1265
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subjects Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, Ion Exchange - methods
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Geotrichum - enzymology
Geotrichum - growth & development
Hydrolases
Indicators and Reagents
Kinetics
Lipase - analysis
Lipase - isolation & purification
Lipase - metabolism
Oleic Acid
Oleic Acids
Rhodamines
Sepharose
Spectrometry, Fluorescence - methods
Substrate Specificity
Triglycerides - metabolism
title Determination of Lipase Activity by a Rhodamine-Triglyceride-Agarose Assay
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