Determination of di-/Polyamine Oxidase Activity in Plants by an In-Gel Spermidine Oxidation Assay

Methods Mol Biol. 2018;1694:141-147. doi: 10.1007/978-1-4939-7398-9_14.


Diamine and polyamine catabolism controls plant development, resistance to pathogens and stress responses. Diamine and polyamine oxidases control the catabolism of diamines and polyamines, respectively. Two major routes of di-/polyamine catabolism exist: the terminal and the interconverting. The in vitro activity of each route is assayed by the colorimetric or chemiluminescent determination of hydrogen peroxide produced by oxidation of di-/polyamine substrates. However, these assays fail to estimate activity of individual di-/polyamine oxidase isoenzymes. Herein, I describe an assay for the simultaneous in-gel determination of terminal and interconverting di-/polyamine oxidase isoenzyme activities.

Keywords: Diamine oxidases; Flavin adenine dinucleotide; Hydrogen peroxide; Oxidation; Peroxisomes; Polyamine back-conversion; Polyamine interconversion; Polyamine oxidases; Spermidine.

MeSH terms

  • Enzyme Assays*
  • Hydrogen Peroxide / chemistry
  • Hydrogen Peroxide / metabolism
  • In Vitro Techniques
  • Oxidation-Reduction*
  • Oxidoreductases Acting on CH-NH Group Donors / metabolism*
  • Plant Extracts / chemistry
  • Plants / enzymology*
  • Spermidine / chemistry*


  • Plant Extracts
  • Hydrogen Peroxide
  • Oxidoreductases Acting on CH-NH Group Donors
  • polyamine oxidase
  • Spermidine