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.