title: Purification and properties of fructosyl lysine oxidase from Fusarium oxysporum S-1F4.
abstract: Fructosyl lysine oxidase (FLOD) was examined for its use in the enzymatic measurement of the level of glycated albumin in blood serum. To isolate microorganisms having such an enzyme activity, we used N epsilon-fructosyl N alpha-Z-lysine (epsilon-FL) as a sole nitrogen source in the enrichment culture medium. The isolated fungus, strain S-1F4, showed a high FLOD activity in the cell-free extract and was identified as Fusarium oxysporum. FLOD was purified to an apparent homogeneity on SDS-PAGE. The molecular mass of the subunit was 50 kDa on SDS-PAGE and seemed to exist in a monomeric form. The enzyme had an absorption spectrum characteristic of a flavoprotein and the flavin was found to be covalently bound to the enzyme. The enzyme acted against N epsilon-fructosyl N alpha-Z-lysine and N alpha-fructosyl N epsilon-Z-lysine and showed specificity for fructosyl lysine residues.
authors.affiliation: Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Sakai
authors.forename: N
authors.initials: N
authors.lastname: Yoshida
authors.forename: A
authors.initials: A
authors.lastname: Isogai
authors.forename: Y
authors.initials: Y
authors.lastname: Tani
authors.forename: N
authors.initials: N
authors.lastname: Kato
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000645:Ammonium Sulfate; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D005415:Flavins / Q000032:analysis; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism; D005670:Fusarium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D005415:Flavins; D005656:Fungal Proteins; D000594:Amino Acid Oxidoreductases; C093463:fructosyl lysine oxidase; D000645:Ammonium Sulfate