Storage and Maintaining Activity of Ribulose Bisphosphate Carboxylase/Oxygenase

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Abstract
Purified ribulose-1,5-bisphosphate carboxylase/oxygenase in 50% saturated (NH4)2SO4 was stable when frozen as small beads in liquid nitrogen and stored at −80 C. When stored as a slurry at 4 C most of the activity was lost within four weeks. This loss was due not only to enzyme polymerization. Activity in old preparations purified from spinach leaves, but not tobacco or tomato leaves, can be restored to the level of newly purified enzyme after storage at 4 C by treatment with 50 to 100 millimolar dithiothreitol for several hours followed by dialysis against buffer and 1 millimolar dithiothreitol before CO2 and Mg2+ activation and assay. Some enzyme oligomers that had been formed were not converted back to native enzyme by treatment with 100 millimolar dithiothreitol.