title: Purification and characterization of a lipase from Aspergillus oryzae.
abstract: A lipase from Aspergillus oryzae was purified by ammonium sulfate fractionation, anion exchange chromatography, hydrophobic interaction chromatography, and anion exchange chromatography. The purified enzyme was a monomeric protein with a molecular mass of 41 kDa estimated by SDS-PAGE and 39 kDa by gel filtration. The optimum pH at 30 degrees C and optimum temperature at pH 7.0 were 7.0 and 30 degrees C, respectively. The enzyme was stable over a pH range of 6-9 at 25 degrees C for 18 h, and up to 30 degrees C at pH 7.0 for 3 h. Ag+, Fe3+, Hg2+, Cu2+, and Zn2+ inhibited the enzyme activity severely. The enzyme was a lipase that hydrolyzed monoacylglycerols and diacylglycerols, but did not hydrolyze triacylglycerols. The N-terminal amino acid sequence of the enzyme was highly homologous with that of the mono- and diacylglycerol lipase from Penicillium camembertii U-150.
authors.affiliation: Food Technology Research Institute of Nagano Prefecture, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Toida
authors.forename: K
authors.initials: K
authors.lastname: Kondoh
authors.forename: M
authors.initials: M
authors.lastname: Fukuzawa
authors.forename: K
authors.initials: K
authors.lastname: Ohnishi
authors.forename: J
authors.initials: J
authors.lastname: Sekiguchi
mesh_terms: D000595:Amino Acid Sequence; D000645:Ammonium Sulfate; D001236:Aspergillus oryzae / Q000201:enzymology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000737:chemistry; D007202:Indicators and Reagents; D008049:Lipase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D008670:Metals / Q000737:chemistry / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity
chemical_list: D005227:Fatty Acids; D007202:Indicators and Reagents; D008670:Metals; D008049:Lipase; D000645:Ammonium Sulfate