Catalytic Reaction and Detection Principle of Glutamate Dehydrogenase GLDH

Glutamate dehydrogenase (GLDH) or GDH is an enzyme related to amino acid metabolism in vivo, present in liver, brain, and kidney tissues, and is commonly used as a detection indicator for liver diseases. GLDH can catalyze the dehydrogenation of L-glutamic acid to produce alpha ketoglutaric acid, which enters the tricarboxylic acid cycle.

Glutamic acid is an acidic amino acid with two carboxyl groups in its molecule. Its chemical name is alpha aminoglutarate, and it plays an important role in protein metabolism in organisms. It participates in many important chemical reactions in animals, plants, and microorganisms. Glutamic acid (2-aminoglutaric acid) has three isomers, the left isomer, the right isomer, and the racemic isomer. L-Glutamic acid is its left-handed glutamate, and the glutamate dehydrogenase capable of dehydrogenation (oxidative deamination) is L-Glutamic acid.

Glutamate dehydrogenase

Glutamate dehydrogenase is mainly present in the mitochondria of liver, heart, and kidney cells, with small amounts found in the mitochondria of brain, skeletal muscle, and white blood cells. In addition to catalyzing the reversible reaction of L-glutamic acid dehydrogenation, it also has the function of catalyzing other amino acids, such as catalyzing the oxidation of L-valine, L-2-aminobutyric acid, and L-leucine deamination.

Principle of Glutamate Dehydrogenase GDH Detection:

The activity detection principle of glutamate dehydrogenase GLDH finished product reaction equation: α - ketoglutarate α - ketoglutarate+NH3+NADH+H+is catalyzed by glutamate dehydrogenase to generate glutamate GLDH+NAD++H2O NADH. Reagent: A 0.1 M Tris HCl buffer, pH 8.3 B 1.5M NH4Cl solution C 0.225M α - ketoglutaric acid solution (pH 7.0-9.0) D 7.5mM NADH solution E Enzyme diluent: 0.1 M Tris HCl buffer, pH 8.3. The consumption of NADH can be detected by measuring the light absorption at 340nm. Measurement time: 180s Delay time: 60s Integration time: 120s Coefficient/factor: 6.776 Measurement temperature: 30 ± 1 ℃ 3.2 Sample preparation If the sample to be tested is solid, it can be dissolved in a ratio of 10mg sample/1000ul ultrapure water. Dissolve and place at 2-8 degrees Celsius.

Glutamate dehydrogenase is a very important enzyme in the liver, involved in the metabolism of glutamate, alpha ketoglutarate, and other amino acids. Therefore, the detection of serum GDH content is an important indicator of whether the body is healthy. The GDH enzyme preparation provided by Desheng is the raw material of the reagent kit, mainly used for biochemical detection and related research experiments.

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