Purification and Properties of FructosylLysine 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 micoorganisms having such an enzyme activity, we used N^ε-fructosyl N^α-Z-lysine (ε-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^ε-fructosyl N^α-Z-lysine and N^α-fructosyl N^ε-Z-lysine and showed specificity for fructosyl lysine residues.