PLoS ONE (Jan 2023)

Purification and characterization of novel isoforms of the polyphenol oxidase from Malus domestica fruit pulp.

  • Naila Sajjad,
  • M Sheeraz Ahmad,
  • Raja Tahir Mahmood,
  • Muhammad Tariq,
  • Muhammad Javaid Asad,
  • Shamaila Irum,
  • Anisa Andleeb,
  • Abid Riaz,
  • Dawood Ahmed

DOI
https://doi.org/10.1371/journal.pone.0276041
Journal volume & issue
Vol. 18, no. 8
p. e0276041

Abstract

Read online

Polyphenol oxidases (PPOs), belong to the group of oxidoreductases that are copper containing enzymes and are responsible for plant browning. PPOs are extensively distributed in plant kingdom and can oxidize wide range of aromatic compounds of industrial importance. The aim of this study was purification and characterization of PPO isoforms from the fruit pulp of Golden delicious apple. High performance liquid chromatography was used to purify the two novel isoforms of PPO and further their molecular weights (45 and 28 kDa) were determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified isoforms have optimum pH (6.5), optimum temperature (40°C), the Vmax (4.45 μM/min) and Km (74.21 mM) with catechol substrate. The N-terminal microsequences of both PPO isoforms were determined using a pulse liquid protein sequencer and found to be AKITFHG (28 kDa) and APGGG (45 kDa). Polyphenol oxidases are efficiently used in the pharmaceutical, paper and pulp, textiles and food industries. Recently, the PPOs have been used for bioremediation and in the development of biosensors.