Use of naturally fluorescent triacylglycerols from Parinari glaberrimum todetect low lipase activities from Arabidopsis thaliana seedlings

The aim of this study was to design a convenient, specific, sensitive, and continuous lipase activity assay using natural long-chain triacylglycerols (TAGs), Oil was extracted from Parinari glaberrimum seed kernels and the pu rified TAGs were used as a substrate for detecting low levels of Lipase act ivities. The purified TAGs are naturally fluorescent because more than half of the fatty acids from Parinari oil are known to contain 9,11,13,15-octad ecatetraenoic acid (parinaric acid) in its esterified form, The presence of detergents (sodium taurodeoxycholate, CHAPS, Sulfobetaine SB12, Tween(R) 2 0, Brij(R) 35, Dobanol(R), n-dodecylglucoside) above their critical micella r concentration dramatically increases the fluorescence of the parinaric ac id released by various lipases. This increase in the fluorescence intensity is linear with time and proportional to the amount of lipase added. This n ew method, performed under non-oxidative conditions, was applied successful ly to detecting low lipase levels include protein extracts from plant seeds and could. be scaled down to microtiterplate measurements. Quantities as l ow as 0.1 ng of pure pancreatic Lipase could be detected under standard con ditions (pH 8), Lipase activity can also be assayed in acidic media (pH 5) using human gastric lipase, This simple and continuous assay is compatible with a high sample throughput and might be applied to detecting true lipase activities in various biological samples.-Beisson, F., N. Ferte,J. Nari, G . Neat, V. Arondel, and R. Verger. Use of naturally fluorescent triacylglyc erols from Parinari glaberrimum to detect low Lipase activities from Arabid opsis thaliana seedlings.