Purification of rat pancreatic lipase

Abstract

A procedure for the isolation of lipase (glycerolester hydrolase, EC 3.1.1.3) from rat pancreas is described. The purification scheme includes homogenization of the pancreas. centrifugation at 3,000 rpm, centrifugation at 40,000 rpm. DEAE-cellulose chromatography, precipitation of amylase as the amylase-glycogen complex, gel filtration of the amylase-free proteins on Sephadex G-100, and chromatography on carboxymethyl-Sephadex C-50. The enzyme showed only one band on polyacrylamide gel electrophoresis and had a specific activity of 5330 ± 80 units/mg of protein.

Keywords

• amylase
• DEAE-cellulose
• Sephadex G-100
• CM-Sephadex C-50