10] Purification of carboxyl ester lipase (bile salt-stimulated lipase) from human milk and pancreas

Abstract

Publisher Summary This chapter summarizes the physiological motivation for the interest in the purification of the human enzyme. Purification of carboxyl ester lipase is a story of purification of several enzymes using different substrates. Thus, several strategies and methods have been used to obtain a pure enzyme. It has been determined that all these enzymes are the same and are found in both human milk and pancreas. Purification of the enzyme from pancreatic gland is more cumbersome, which may explain why there is just one published study from 1988 available for the human enzyme. This purification scheme includes homogenization of the pancreatic tissue in the presence of trypsin inhibitor and bile salts, centrifugations, ammonium sulfate precipitation, filtration, cholate-Sepharose chromatography, and Bio-Rex and heparin-Sepharose chromatography. Purification of the milk enzyme is easier, owing to the presence of fewer proteases. The chapter presents different strategies for purification of the human enzyme from milk and pancreas, owing to the significant difference in the source of the enzyme—milk or pancreatic juice/pancreas.