Isolation of Intact Chloroplasts from Spinach Leaf by Centrifugation in Gradients of the Modified Silica “Percoll”

Abstract

The isolation of the photosynthetically competent chloroplast preparations was undertaken by means of the density gradient centrifugation on the modified silica sol “Percoll.” A clear separation of the intact chloroplast sustaining the high photosynthetic activities (light dependent CO² fixation ca. 130 μmol/mg Chl•hr) was established. The contamination of mitochondria and peroxisomes was estimated to be less than 3% by measuring the activities of their marker enzymes. The chloroplasts were proved to be free from endoplasmic reticulum and cytosol. The photosynthetic CO² fixation of the isolated chloroplast preparations was saturated by illumination of the light intensity of 20, 000 Lux (12 mW/cm², 400_??_750nm).