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Glutamate synthase (GOGAT) is widely distributed in plants, and 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 glutamate synthase (GOGAT). 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).
NADH-glutamate synthase (NADH-GOGAT, EC 1.4.1.14) catalyzes the transfer of the amino group from glutamine to α-ketoglutarate, forming two molecules of glutamate; simultaneously, NADH is oxidized to NAD+. The enzyme activity of NADH-GOGAT can be determined by measuring the rate of decrease in absorbance at 340 nm.
The reaction catalyzed by this enzyme is: L-glutamine + 2-oxoglutarate + NADH + H+ = 2 L-glutamate + NAD+
Additional Materials and Equipments Required
UV spectrophotometer, 1mL quartz cuvettes (optical path 1cm), desktop centrifuge, water bath, pipettes, mortar and pestle, ice, and distilled water.