I am trying to find lipase activity in mustard seeds. I developed a method by potentiometry but i am not getting repeatability. Is there any other method to estimate lipase activity.
Dear Phani Buggala Use most reliable spectroscopic method for lipase activity determination in which Lipase activity was assayed with in the medium broth test samples by the method of Winkler and Stuckmann 1979 by measuring the micromoles of p-nitrophenol (p-NP) discharged from p-nitrophenylpalmitate (p-NPP).
Molecular characterization and bioinformatics studies of a lipase from Bacillus thermoamylovorans BHK67, International J. Biol. Macromol. 107(2017) 2131-2140.
If you not get result go through as suggested by Prof. Kou Hayakawa.
Dear Phani Buggala Use most reliable spectroscopic method for lipase activity determination in which Lipase activity was assayed with in the medium broth test samples by the method of Winkler and Stuckmann 1979 by measuring the micromoles of p-nitrophenol (p-NP) discharged from p-nitrophenylpalmitate (p-NPP).
Molecular characterization and bioinformatics studies of a lipase from Bacillus thermoamylovorans BHK67, International J. Biol. Macromol. 107(2017) 2131-2140.
If you not get result go through as suggested by Prof. Kou Hayakawa.
Exept for the HPLC method suggested by Prof Hayakawa and the p-nitrophenolic assay suggested by Abhishek Sharma there exists also a turbidimetric assay to determin lipase activity. The principle of the turbidimetric assay is the liberation of free fatty acids from Tween by a lipase and a precipitation of these free fatty acids with Ca ions. It is measured photometrically at 500nm. Maybe this assay could also be an alternative to determine the acitivity of your lipase.