Lipase acts on triacylglycerols and releases fatty acids. Determination of its activity is calculated by titration of free fatty acids.
Substrate preparation
- A buffer solution containing Na2HPO4/NaH2PO4 50 mM, pH=7, is prepared.
- Next an olive oil emulsion (40 ml of olive oil are added to 60 ml of gum arabic – emulsifier solution 5%, w/v) is prepared and the mixture is homogenized in laboratory homogenizer.
- The substrate of the enzyme is composed of 50 ml of olive oil emulsion in 45 ml of buffer solution.
Determination of enzymatic activity
- 0.5 ml of crude enzyme solution is added into 9.5 ml of substrate.
- The mixture is incubated in a shaker for 1 h at T=28 °C
- This is followed by titration of the mixture with NaOH solution 50 mM until pH=9
A unit of activity of lipase is the quantity of the enzyme that releases 1μmole of fatty acids in 1 h at T=28 °C.