Biochemical Studies on Lipase Isolated from Camel Pancreas

Abstract

This investigation aimed to establish a simple and reproducible procedure for isolation and purification of pancreatic lipase (glycerol ester hydrolase EC 3.1.1.3) from camel pancreas and to study its physical and chemical properties whereas this enzyme is used as a digestive drug in case of acute pancreatities.

Isolation and purification process includes:

(1) Homogenization of camel pancreas by a mechanical means.

(2) Complete dehydration and defating using organic solvents (acetone and trichloroethylene) to obtain dry defated pancreatic powder (crude powder).

(3) The crude powder was extracted in different buffers with different ionic strength at different pH's until the best method of extraction was obtained (gave the highest activity with good yield which was found to be the extraction of the crude powder in O.OIM tris-HCl buffer, pH 7, containing 4 mM CaCh.

(4) Fractionation with ammonium sulfate at 20%-80% saturation.

(5) Chromatography on DEAE-cellulose column where lipase fraction was obtained as a single peak at O.I-0.125M NaCl.

(6) The final purification step was the gel filtration of lipase fraction obtained from DEAE-cellulose chromatography step, on Sephadex G- 100 where this step increases the specific activity of the enzyme to 2298.5 units/mg protein with olive oil as substrate and to 4597 units/mg protein with tributyrin as substrate in the presence of 3 mM