Glucokinase (GCK) is a key enzyme in the glucose metabolism pathway. It participates in the first step of glycolysis, catalyzes glucose phosphorylation, promotes insulin secretion and glucose metabolism, and effectively controls blood glucose balance in internal. GCK, also known as hexokinase IV, is mainly expressed in hepatocytes and islet β-cell, and has a stronger affinity for glucose than other hexokinases. Therefore, the total hexokinase activity in the sample is measured at high glucose concentration, and the hexokinase activity with low glucose affinity is measured at low glucose concentration. The GCK activity in the sample can be obtained by subtracting the total hexokinase from the total hexokinase. The detection principle of this kit: GCK catalyzes the substrate reaction to produce NADPH, which reacts with the chromogenic agent. The activity of GCK can be calculated by measuring the generation rate of chromogenic substance at 450 nm.
Under the presence of G6PDH, glucose-6-phosphoric acid is oxidized to 6-PG. In this reaction, NADP+ is reduced to NADPH. Under the action of electron coupling reagent 1-MPMS, NADPH reducts wST-8 to form orange formazan, which has the maximum absorption peak at about 450 nm. Formazan generated in the reaction system is proportional to the activity of G6DPH in the sample.
Glucose-6-phosphate (G6P) is a molecule generated by phosphorylation of hydroxyl groups on the sixth carbon of glucose under the catalysis of hexokinase. It is a common small molecule of sugar metabolism in cells and participates in biochemical pathways such as glycolysis and pentose phosphate pathway. In the first reaction of glycolysis, glucose is catalyzed by hexokinase to produce glucose-6-phosphate, which is then catalyzed by phosphoglucose isomerase to form fructose-6-phosphate to continue the other steps of glycolysis: In the pentose phosphate pathway, glucose-6-phosphate is the first substrate, and this process is also the main way to generate NADPH. In addition to these two metabolic pathways, glucose-6-phosphate can also be converted into glycogen or starch and stored.
Glucose oxidase can catalyze the oxidation of glucose to gluconic acid to produce hydrogen peroxide. In the presence of chromogenic oxygen receptors, peroxidase catalyzes hydrogen peroxide and oxidizes pigment sources to form colored substances. Measure the OD value at 505 nm and glucose content can be calculated indirectly.
Glucose oxidase can catalyze the oxidation of glucose into gluconic acid and produce hydrogen peroxide. In the presence of peroxidase, hydrogen peroxide reacts with the non-fluorescent substance to form fluorescent substance. The glucose content can be calculated indirectly by measuring the fluorescence intensity at the excitation wavelength of 535 nm and the emission wavelength of 590 nm.
Glucose oxidase can catalyze the oxidation of glucose to produce hydrogen peroxide. In the presence of peroxidase, hydrogen oxidizes pigment sources to form colored substances. Measure the OD value at 550 nm and glucose oxidase (GOD) activity can be calculated indirectly.
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