An ultraviolet spectrophotometric assay for measuring lipase activity using long-chain triacyglycerols from Aleurites fordii seeds

Analytical Biochemistry
Gaëlle Pencreac'hR Verger

Abstract

In this study, we designed a specific, continuous, and sensitive UV spectrophotometric lipase assay using natural triacylglycerols (TAGs) from the Aleurites fordii seed oil (tung oil). alpha-Eleostearic acid (9,11,13-cis, trans,trans-octadecatrienoic acid) is the main fatty acid component (it accounts for up to 70%) of the TAGs from tung oil. The conjugated triene present in alpha-eleostearic acid constitutes an intrinsic chromophore, which confers strong UV absorption properties on both the free fatty acid and the TAGs from tung oil. The lipase assay is based on the difference between the apparent molar extinction coefficients of the two types of alpha-eleostearic acid present, that which is esterified into TAGs and that which is released into the reaction medium. This difference is responsible for the variations in the UV absorption spectrum of the reaction medium occurring upon enzymatic TAGs hydrolysis. Using the purified lipase from Thermomyces lanuginosa (TLL) and the detergent sodium taurodeoxycholate (NaTDC, 4 mM), it was established that the most suitable method of measuring lipolysis consisted of monitoring the decrease in the OD at 292 nm, which was linear with time and proportional to the amount of lipase added. In ...Continue Reading

References

May 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·L A SklarR D Simoni
Jan 7, 1997·Biochimica Et Biophysica Acta·Y GargouriR Verger
Jul 23, 1999·Annals of the New York Academy of Sciences·F H ArnoldK Miyazaki
Jul 3, 1998·Angewandte Chemie·Rolf D Schmid, Robert Verger

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Citations

Sep 8, 2015·Biochimie·Serdar ÜlkerJean-François Cavalier
May 20, 2016·Journal of Lipid Research·Meddy El AlaouiAbdelkarim Abousalham
Mar 12, 2019·Chembiochem : a European Journal of Chemical Biology·Serina L RobinsonLawrence P Wackett
Dec 13, 2019·Journal of Ethnobiology and Ethnomedicine·Fengke LinChunlin Long
Dec 9, 2014·Chemical Biology & Drug Design·Karima LahbibSoufiane Touil
Sep 18, 2020·Biochimie·Milena Chagas LisboaCleide Mara Faria Soares

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