title: Purification and characterization of triacylglycerol lipase from Aspergillus oryzae.
abstract: Triacylglycerol lipase (L3) was purified from Aspergillus oryzae RIB128 by ammonium sulfate fractionation, acetone precipitation, anion-exchange chromatography, and gel filtration. The purified enzyme was formed from a glycoprotein and a monomeric protein with molecular masses of 25 and 29 kDa, by SDS-PAGE and gel filtration, respectively. The optimum pH at 40 degrees C was 5.5 and the optimum temperature at pH 5.5 was 40 degrees C. The enzyme was stable between a pH range of 4.0-7.5 at 30 degrees C for 24 h, and at up to 30 degrees C at pH 5.5 for 1 h. Heavy metal ions, detergents, DFP, and DEP strongly inhibited the enzyme activity. The lipase hydrolyzed not only triacylglycerols but also monoacylglycerols and diacylglycerols. The enzyme had higher specificity toward triacylglycerols of middle-chain saturated fatty acids than short-chain or long-chain fatty acids. The enzyme had 1,3-positional specificity. The N-terminal amino acid sequence of the enzyme was not significantly similar to that of other lipases with published sequences.
authors.affiliation: Food Technology Research Institute of Nagano Prefecture, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Toida
authors.forename: Y
authors.initials: Y
authors.lastname: Arikawa
authors.forename: K
authors.initials: K
authors.lastname: Kondou
authors.forename: M
authors.initials: M
authors.lastname: Fukuzawa
authors.forename: J
authors.initials: J
authors.lastname: Sekiguchi
mesh_terms: D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology*; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D008049:Lipase / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D007202:Indicators and Reagents; D008670:Metals; D008049:Lipase