Regeneration of denatured polyphenol oxidase in Toona sinensis (A.Juss.) Roam

The regeneration of polyphenol oxidase (PPO) from leaves of Toona sinensis (A.Juss.) Roam (TS), denatured by boiling with 0.4% (w/v) sodium dodecyl sulfate (SDS) and 1% (v/v) β-mercaptoethanol, and was studied with SDS-polyacrylamide gel electrophoresis (PAGE) and the gel-activity-stained assay. The optimal regenerating conditions for the denatured TS-PPO were as follows: extracting with buffer of 150 mM Na-Pi, pH 7.2; regenerating with buffer of 50 mM Tris–HCl, pH 7.2, 2% Triton X-100 for 45 min. In addition, the PPO regeneration was promoted by the addition of Cu2+ in the recovery buffer. The regenerating activity of the denatured-PPO was reduced in samples extracted with buffers containing polyvinylpyrrolidone (PVP), polyvinylpolypyrrolidone (PVPP) or Triton X-100.