CHARACTERIZATION OF GRAPEFRUIT FLAVEDO PEROXIDASES

Abstract

Grapefruit (Citrus paradisi) flavedo is a rich source of peroxidase (POD) (EC 1.11.1.7). Changes in POD have been related to senesence and environmental stress in a variety of plant tissues. However, due to the large number of POD isoenzymes as well as the broad substrate specificity, measurement of POD activity in crude extracts is of limited value for gaining an understanding of the role of POD in vivo. We have begun to purify and characterize POD isoenzymes from grapefruit flavedo. HPLC gel permeation chromatography reveals 2 peaks of POD activity with apparent MW of 66 kD and 30 kD. Native PAGE (8% bis-acrylamide, pH 8.8) followed by activity staining indicates that the PODs differ in Pi; the 30 kD POD migrates anodally, whereas the 66 kD POD does not migrate. Isoelectric focusing has been used to separate flavedo PODs into acid (Pi ca 4.0) and basic (Pi > 8.5) forms. Treatment of grapefruit with ethylene (2 ppm 72 hours) induces a basic POD not present in freshly-harvested fruit or in nonethylene-treated controls.