Dromedary pancreatic lipase: purification and structural properties.
Dromedary pancreatic lipase was purified from delipidated pancreases. Pure dromedary pancreatic lipase (glycerol ester hydrolase, EC 3.1.1.3) was obtained after ammonium sulfate fractionation, Sephadex G-100 gel filtration, anion-exchange (Mono Q Sepharose) and size exclusion column using high performance liquid chromatography (HPLC). The pure lipase is a monomer and has a molecular mass of about 45 kD and a pI of around 4.8. A specific activity of 5900 U/mg was measured on tributyrin as substrate at 37 degrees C in the presence of colipase and 2 mM NaTDC. The first 11 N-terminal amino acid residues and 10 peptides obtained by endoproteinase Glu-C digestion were sequenced. Dromedary pancreatic lipase is very similar to other pancreatic lipases as compared with their N-terminal and some peptides sequences. DrPL is activated by interfaces. The interfacial activation could be related to the presence of a lid and in fact one fragment of this lid domain (P9) was sequenced here: its' role will be discussed below.
Laboratoire de Biochimie, ENIS, Sfax, Tunisia.
H
H
Mejdoub
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J
Reinbolt
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Y
Gargouri
D000595:Amino Acid Sequence; D000818:Animals; D002162:Camelus / Q000378:metabolism*; D002417:Cattle; D002645:Chickens; D004285:Dogs; D004789:Enzyme Activation; D006041:Goats; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D014421:Turkey
D008049:Lipase