Abstract
The plant Cleome gynandra L. of the family Capparaceae was studied for the localization of two photosynthetic carboxylating enzymes, phosphoenolpyruvate (PEP) carboxylase (EC 4.4.4.31) and ribulose 1,5-bisphosphate (RuBP) carboxylase (EC 4.1.1.39). In preliminary studies, the plant showed Kranz-type leaf anatomy and higher activity of PEP carboxylase in whole leaf extracts than the species of this genus. The mesophyll protoplasts (MP) and bundle sheath protoplasts (BSP) from leaves of C. gynandra were isolated, purified, mechanically ruptured and chloroplasts were separated by differential centrifugation. The protoplasts were found intact based on the exclusion of Evan's Blue under light microscope. PEP carboxylase was exclusively found in the non-particulate fraction after differential centrifugation of mesophyll protoplast extract and RuBP carboxylase in chloroplasts of bundle sheath protoplasts. The results suggest that PEP carboxylase and RuBP carboxylase are localized in the cytoplasm of mesophyll cells and bundle sheath chloroplasts respectively.
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