RuBP carboxylase determination by enzymic estimation of D-3-PGA formed.

RuBPcarboxylase activity was measured in extracts of barley (Hordeum Vulgare L., cv. HOP) seedlings both with the standard radiometric method and by measuring D-3-phosphoglyceric acid formed enzymically in a two stage assay. In the different conditions used, characterized by different NaHCO3 concentrations, different pH and the presence and absence of oxygen, essentially the same ratio of D-3-PGA formed per (14)CO2 fixed was obtained. This ratio respected the known stoichiometry of two molecules of D-3-PGA formed per CO2 fixed.It is suggested that measurement of D-3-PGA enzymically in a two stage assay can be routinely used for the determination of RuBP case activity instead of the radiometric method. The advantages and the validity of the method are discussed.