Determination of lipase activity in cheese using trivalerin as substrate

Lipolysis during cheese ripening is usually assessed by the accumulation of free fatty acids (FFA). An assay to determine total lipolytic activity present in a cheese during ripening was established. Finely grated cheese (1 g) was directly incubated with trivalerin (204 mg) as a substrate at 35°C for 4 h. Free valeric acid was extracted and quantified by gas chromatography. The assay was linear with time up to 24 h and up to 0.11 Lipase Units g^-1 cheese. The total amount of lipolytic activity determined with this assay was consistent with the total amount of FFA present in 5 types of cheese. In Idiazabal cheese samples manufactured with or without lipase added, the total lipolytic activity determined during ripening remained constant up to 6 months of ripening. In addition to trivalerin, other possible substrates investigated were triolein, triundecanoin and the endogenous butyric acid-containing triacylglycerols present in the cheese sample. Activities measured with these substrates were considerably lower than values obtained with trivalerin due to the high levels of oleic acid present in cheese, or to difficulties in mixing triundecanoin (solid at 35°C) with the grated cheese sample. Endogenous triacylglycerols gave increasingly lower activity values as ripening time progressed due to substrate depletion.