Glutamate synthase in Medicago sativa L. occurrence and properties of Fd-dependent enzyme in plant cell fraction during root nodule development

Abstract

In the plant cell fraction of Medicago sativa (L. cv Europe) nodules, glutamate synthase is active with reduced Fd, MV, NADH and NADPH as an electron donor. Up to 25 to 30 days after inoculation, the activities of Fd-dependent glutamate synthase (EC 1.4.1.7), the most active form of the enzyme, NADH-dependent (EC 1.4.1.14) and NADPH-dependent (EC 1.4.1.13) glutamate synthases increase about 2-fold followed by a relatively constant level per gram fresh weight of nodules. The activities of glutamate synthases with different electron carriers increase constantly about 30-fold after 46 days of inoculation by total fresh weight of nodules per plant. These nodule glutamate synthase activities with Fd, NADH or NADPH represent 30% relative to those of root glutamate synthases per plant with the respective electron donor. Fd-glutamate synthase in nodule plant fraction is a protein molecule immunochemically distinct from pyridine nucleotide-glutamate synthases. MV-linked enzyme activity is associated with Fd-glutamate synthase. The Fd-glutamate synthase has a subunit molecular mass of 68.2 kDa, and it exhibits a high affinity for spinach Fd as an electron carrier. The increase in Fd-glutamate synthase activity during nodule development is accompanied by a rise in the enzyme protein content. The total activity of different forms of glutamate synthase in vitro ensures a higher level than the rate of ammonia production during N2 fixation in bacteroids of Medicago sativa nodules.