Abstract
Three C 4 species of the genera Brachiaria and Urochloa from the family Gramineae and 4 CAM species from the families Asclepiadaceae, Bromeliaceae, and Liliaceae were studied for the intracellular localization of the decarboxylating enzyme phosphoenolpyruvate (PEP) carboxykinase. Bundle sheath protoplasts of the C 4 plants and mesophyll protoplasts of the CAM plants were isolated, purified, mechanically ruptured, and organelles separated by differential centrifugation. The chloroplasts were about 70 95% intact based on the retention of chloroplast marker enzymes. Most of the PEP carboxykinase was in the non-chloroplastic fraction (98 100% in C 4 and 80% or more in CAM). The enzyme was still largely non-particulate after centrifugation of protoplast extracts of Urochloa mosambicensis (C 4) and Ananas comosus (CAM) to pellet chloroplasts, mitochondria, and microbodies. These results suggest that PEP carboxykinase is localized mainly in the cytoplasm in both C 4 and CAM plants.
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