The oxidase catalyzes the transfer of four electrons from reduced plastoquinone to molecular oxygen to form water . The net reaction is written below:
2 QH + O → 2 Q + 2 HO
Analysis of substrate specificity revealed that the enzyme almost exclusively catalyzes the reduction of plastoquinone over other quinones such as ubiquinone and duroquinone. Additionally, iron is essential for the catalytic function of the enzyme and cannot be substituted by another metal cation like Cu, Zn, or Mn at the catalytic center.
It is unlikely that four electrons could be transferred at once in a single iron cluster, so all of the proposed mechanisms involve two separate two-electron transfers from reduced plastoquinone to the di-iron center. In the first step common to all proposed mechanisms, one plastoquinone is oxidized and both irons are reduced from iron(III) to iron(II). Four different mechanisms are proposed for the next step, oxygen capture.  One mechanism proposes a peroxide intermediate, after which one oxygen atom is used to create water and another is left bound in a diferryl configuration. Upon one more plastoquinone oxidation, a second water molecule is formed and the irons return to a +3 oxidation state. The other mechanisms involve the formation of Fe(III)-OH or Fe(IV)-OH and a tyrosine radical. These radical-based mechanisms could explain why over-expression of the PTOX gene causes increased generation of reactive oxygen species.