Regulation of Soybean Net Photosynthetic CO 2 Fixation by the Interaction of CO 2 , O 2 , and Ribulose 1,5-Diphosphate Carboxylase

Abstract

Kinetic properties of soybean net photosynthetic CO(2) fixation and of the carboxylase and oxygenase activities of purified soybean (Glycine max [L.] Merr.) ribulose 1, 5-diphosphate carboxylase (EC 4.1.1.39) were examined as functions of temperature, CO(2) concentration, and O(2) concentration. With leaves, O(2) inhibition of net photosynthetic CO(2) fixation increased when the ambient leaf temperature was increased. The increased inhibition of CO(2) fixation at higher temperatures was caused by a reduced affinity of the leaf for CO(2) and an increased affinity of the leaf for O(2). With purified ribulose 1,5-diphosphate carboxylase, O(2) inhibition of CO(2) incorporation and the ratio of oxygenase activity to carboxylase activity increased with increased temperature. The increased O(2) sensitivity of the enzyme at higher temperature was caused by a reduced affinity of the enzyme for CO(2) and a slightly increased affinity of the enzyme for O(2). The similarity of the effect of temperature on the affinity of intact leaves and of ribulose 1,5-diphosphate carboxylase for CO(2) and O(2) provides further evidence that the carboxylase regulates the O(2) response of photosynthetic CO(2) fixation in soybean leaves. Based on results reported here and in the literature, a scheme outlining the stoichiometry between CO(2) and O(2) fixation in vivo is proposed.Oxygen competitively inhibited carboxylase activity with respect to CO(2), and CO(2) competitively inhibited oxygenase activity with respect to O(2). Within the limits of experimental error, the Michaelis constant (CO(2)) in the carboxylase reaction was identical with the inhibition constant (CO(2)) in the oxygenase reaction, and the Michaelis constant (O(2)) in the oxygenase reaction was identical with the inhibition constant (O(2)) in the carboxylase reaction. The Michaelis constant, (ribulose 1,5-diphosphate) was the same in both the carboxylase and oxygenase reactions. This equality of kinetic constants is consistent with the notion that the same enzyme catalyzes both reactions.