Characterization of turkey pancreatic lipase.
Turkey pancreatic lipase (TPL) was purified from delipidated pancreases. Pure TPL (glycerol ester hydrolase, EC 3.1.1.3) was obtained after ammonium sulfate fractionation, Sephacryl S-200 gel filtration, anion exchange chromatography (DEAE-Sepharose) and size exclusion column using high performance liquid chromatography system (HPLC). The pure lipase, which is not a glycoprotein, was presented as a monomer having a molecular mass of about 45 kDa. The lipase activity was maximal at pH 8.5 and 37 degrees C. TPL hydrolyses the long chains triacylglycerols more efficiently than the short ones. A specific activity of 4300 U/mg was measured on triolein as substrate at 37 degrees C and at pH 8.5 in the presence of colipase and 4 mM NaTDC. This enzyme presents the interfacial activation when using tripropionin as substrate. TPL was inactivated when the enzyme was incubated at 65 degrees C or at pH less than 5. Natural detergent (NaTDC), synthetic detergent (Tween-20) or amphipatic protein (beta-lactoglobulin A) act as potent inhibitors of TPL activity. To restore the lipase activity inhibited by NaTDC, colipase should be added to the hydrolysis system. When lipase is inhibited by synthetic detergent or protein, simultaneous addition of colipase and NaTDC was required to restore the TPL activity. The first 22 N-terminal amino acid residues were sequenced. This sequence was similar to those of mammal's pancreatic lipases. The biochemical properties of pancreatic lipase isolated from bird are similar to those of mammals.
Sayari
A
A
Laboratoire de lipolyse enzymatique, ENIS, BPW 3038, Sfax, Tunisia.
Mejdoub
H
H
Gargouri
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001647:Bile Acids and Salts; D002162:Camelus; D002417:Cattle; D011232:Chemical Precipitation; D002645:Chickens; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods; D003089:Colipases / Q000378:metabolism; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D012756:Sheep; D013696:Temperature; D014422:Turkeys
D001647:Bile Acids and Salts; D003089:Colipases; D008049:Lipase