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Glutamate synthase (GOGAT) is widely distributed in plants. Inorganic nitrogen absorbed by plants, after being reduced to NH4+ by nitrate reductase (NR) and nitrite reductase (NIR), can only be assimilated and utilized through the GS/GOGAT pathway involving glutamine synthetase (GS). GOGAT generally includes two types: one is Fd-GOGAT, which is mostly found in chloroplasts (leaves), and the other is NADH-GOGAT, which is mostly found in proplastids of non-green tissues (roots). Fd-glutamate synthase (Fd-GOGAT, EC 1.4.7.1) catalyzes the transfer of the amino group from glutamine to α-ketoglutarate, forming two molecules of glutamate. Then, a specific enzyme complex decomposes glutamate and reacts with a color developer to form a yellow substance, which has a maximum absorption peak at 450nm, allowing for the determination of Fd-GOGAT enzyme activity.
Additional Materials and Equipments Required
Microplate reader, 96-well plate, desktop centrifuge, water bath, pipettes, mortar and pestle, ice, and distilled water.
