Rapid Two-Phase Extraction of the Calvin Cycle Enzymes Aldolase and Phosphoglycerate Kinase
Fructose-1, 6-bisphosphate aldolase (EC 184.108.40.206) was extracted from isolated spinach chloroplasts in a procedure consisting of two steps; precipitation of bulk protein with polyethylene-glycol (PEG) with Mr = 6500-8000 and partitioning of remaining soluble protein in aqueous two-phase systems. All manipulations, including chloroplast isolation, activity measurements and protein determination, can be carried out within 6 h and the final preparation is 94% pure, on the basis of electrophoretic criteria, with a yield of 19%.
KeywordsLeaf Extract CALVIN Cycle Target Enzyme Phosphoglycerate Kinase CALVIN Cycle ENzyMES
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