GDH (EC 1.4.1.2) is widely distributed in plants. GDH and glutamate synthetase (GOGAT) are involved
in the synthesis of glutamate. It plays an important role in the assimilation of ammonia and the metabolism
of organic nitrogen compounds.
+
GDH catalyzes the NH4 , α-ketoglutaric acid and NADH to form glutamic acid and NAD+, which cause a
decrease in absorbance at 340 nm. The GDH activity is calculated by measuring the rate of decrease in
absorbance at 340 nm.
Reagents and Equipment Required but Not Provided:
Spectrophotometer, desk centrifuge, adjustable pipette, water bath, 1 mL quartz cuvette,
mortar/homogenizer, ice and distilled water.
Sample preparation:
1.Cells or bacterial: Collecting bacteria or cells into a centrifuge tube, discard supernatant after
centrifugation. Suggest 5 million with 1mL of Extract solution. Use ultrasonic to split bacteria or cells
(power 20%, work time 3s，interval 10s，repeat for 30 times). centrifuge at 8000 g for 10 min at 4℃.
Supernatant is placed on ice for test.
2.Tissue: Add 1 mL of Extract solution into 0.1g of tissue, fully grinding on ice. centrifuge at 8000 g for
10 min at 4℃. Supernatant on ice is used for test.
