Purification and characterization of cholesterol esterase from porcine pancreas.
A protein catalyzing the hydrolysis of cholesterol esters and p-nitrophenyl acetate has been purified 200-fold from porcine pancreas. The enzyme is homogenous as judged by polyacrylamide gel electrophoresis and exhibits a molecular weight of 80 000 as determined by sodium dodecyl sulfate electrophoresis and gel filtration. Activity toward p-nitrophenylacetate exhibits a broad pH optimum and is influenced by a group with a pKa of 5.5--6.0. The enzyme is completely inhibited by diisopropylfluorophosphate at concentrations as low as 10(-5) M, suggesting that it is a serine esterase. Partial inhibition was observed with p-chloromercuribenzoate.
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007531:Isoflurophate / Q000494:pharmacology; D008970:Molecular Weight; D009596:Nitrophenols / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D002787:Sterol Esterase / Q000302:isolation & purification / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013552:Swine
D009596:Nitrophenols; D013439:Sulfhydryl Reagents; D007531:Isoflurophate; D002265:Carboxylic Ester Hydrolases; D002787:Sterol Esterase