An improved large scale procedure for the purification of porcine pancreatic lipase

Abstract

A modified procedure for purifying porcine pancreatic lipase (triacylglycerol acyl-hydrolase, EC 3.1.1.3) is described. In comparison to the previous procedure reported by Verger, R., de Haas, G.H.; Sarda, L. and Desnuelle, P. (1969) Biochim. Biophys. Acta 188, 272–282) it is more rapid, more reproducible and results in a purer enzyme preparation. No colipase could be detected in the mixture of isoenzymes and, naturally, in the different separated lipases. In this process, butanol treatment is omitted. After pancreas powder extraction, a batch procedure was used for adsorption of DEAE-cellulose. Sephadex filtration (pH 8.0) was made in a large size column. Finally the isoenzymes were separated on CM-cellulose as in the Verger procedure, but under slightly modified conditions. Lipase L B was fully homogeneous as judged by end group determination, gel electrophoresis (in the presence or absence of sodium dodecyl sulfate) and sedimentation equilibrium.