title: Determination of lipase activity by a rhodamine-triglyceride-agarose assay.
abstract: 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 microgram 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.
authors.affiliation: Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec.
authors.forename: J F
authors.initials: JF
authors.lastname: Jette
authors.forename: E
authors.initials: E
authors.lastname: Ziomek
mesh_terms: D002852:Chromatography, Ion Exchange / Q000379:methods; D005848:Geotrichum / Q000201:enzymology* / Q000254:growth & development; D007202:Indicators and Reagents; D007700:Kinetics; D008049:Lipase / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D019301:Oleic Acid; D009829:Oleic Acids; D012235:Rhodamines; D012685:Sepharose; D013050:Spectrometry, Fluorescence / Q000379:methods; D013379:Substrate Specificity; D014280:Triglycerides / Q000378:metabolism
chemical_list: D007202:Indicators and Reagents; D009829:Oleic Acids; D012235:Rhodamines; D014280:Triglycerides; D019301:Oleic Acid; D012685:Sepharose; D008049:Lipase