Polyamine oxidase from leaves of Lilium longiflorum : Purification and properties

Summary Polyamine oxidase was purified to homogeneity from leaves of Lilium longiflorum using acetone powder, CM-Sephadex and hexamethylenediamine-Sepharose 4B column chromatography. The enzyme showed optimal pH of 5.5 and K m values of 16 and 6μmol · L −1 with spermidine and spermine, respectively. The enzyme was a monomer with molecular mass of 63,000 u (Da) as judged by sodium dodecyl sulfate Polyacrylamide gel electrophoresis. The enzyme appeared to contain 1 mol of flavin adenine dinucleotide per 63,000 g of the enzyme. Hexamethylenediamine was a potent competitive inhibitor with ki of 7 μmol · L −1 . One mol per liter NaCl increased V max of the purified enzyme, similar to polyamine oxidase from oats, and also caused an increase of K m . Antibodies raised against the purified enzyme from Lilium longiflorum did not cross-react with polyamine oxidase from Avena sativa , Eichhonia crassipes , Gibasis pellucida , or Zea mays .