Subcellular localization and isoenzyme pattern of peroxidase and polyphenol oxidase in beet root (Beta vulgaris L.)

The two enzymes involved in enzymatic browning reactions, polyphenol oxidase (PPO) and peroxidase (PO), have been partially purified and extracted from different fractions of beet root. PPO is mainly located in the membrane fraction, and it was also found in the soluble fraction. In both cases PPO was in its latent state. However, PO activity was higher in the soluble fraction than in the membrane fraction. Nevertheless, the highest values of specific activity for PO were obtained from the solubilized enzyme from acetone powders. Under native isoelectric focusing (IEF), several PPO isoenzymes were present in the pH range of 4.8-5.8. All of these isoenzymes shared a single band with a similar apparent mass under sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PO was also analyzed by IEF, showing a complex isoenzyme pattern in all fractions. The characteristic basic PO isoenzyme of high pI found in both the soluble fraction and the solubilized enzyme from acetone powders was not detected in the membrane fraction. The kinetic characterization of PPO and PO from all fractions was carried out.