Pyruvate Kinase Inhibitor Enzyme Preparation EC 2.7.1.40 CAS NO.9001-59-6

In the fields of biochemistry and medical testing, the pyruvate kinase potassium ion assay kit is of crucial importance for accurately detecting pyruvate kinase activity and related potassium ion concentration in biological samples. There are multiple components in this kit that work together to achieve accurate determination of potassium ion concentration.

Acetoacetate kinase substrates and coenzymes

1. Phosphoenolpyruvate (PEP): Phosphoenolpyruvate is an important substrate for pyruvate kinase catalyzed reactions. Under the action of pyruvate kinase, PEP undergoes a phosphate group transfer reaction, transferring the phosphate group to ADP to generate pyruvate and ATP. This reaction is one of the core biochemical reactions in the entire measurement process. The chemical structure of PEP endows it with high reactivity, and its phosphoenolic bond contains a large amount of energy, which can undergo phosphate transfer under the catalysis of pyruvate kinase.

Pyruvate Kinase

2. Adenosine phosphate (ADP): ADP acts as another substrate for the catalytic reaction of pyruvate kinase and participates in the reaction in synergy with PEP. It accepts phosphate groups from PEP and converts them into ATP. The presence of ADP ensures the continuity and integrity of the reaction. The concentration also needs to be accurately controlled, and an appropriate ADP concentration can enable the reaction to proceed at a suitable rate, making it easy to measure accurately. The concentration of ADP affects the equilibrium of the reaction to a certain extent. According to the principle of chemical equilibrium, changing the concentration of ADP can regulate the degree of reaction towards the generation of pyruvate and ATP. In the reagent kit, optimizing the concentration of ADP can ensure that the reaction reaches an appropriate equilibrium state under the measurement conditions, so that the activity of pyruvate kinase can be accurately reflected by the amount of reaction products generated.

3. Reduced Coenzyme I (NADH): After the pyruvate kinase catalyzed reaction, there is usually an indicator reaction to detect the generated pyruvate. NADH participates in this indicator reaction, for example, under the catalysis of lactate dehydrogenase, pyruvate reacts with NADH to produce lactate and oxidized coenzyme I (NAD ⁺). NADH has a specific absorption spectrum, and by detecting the changes in NADH absorption spectrum during the reaction process, the activity of pyruvate kinase can be indirectly determined. The high molar absorptivity of NADH makes it highly sensitive in the detection process. Even trace changes in NADH concentration can be accurately detected by instruments such as spectrophotometers.

The key role of pyruvate kinase

Acetoacetate kinase plays a central role in the entire measurement system. The reaction it catalyzes is a key link in connecting substrates, coenzymes, and subsequent indicator reactions. By catalyzing the reaction between PEP and ADP, not only does it promote the generation of energy metabolism related substances such as ATP, but it also provides a basis for subsequent detection of pyruvate using NADH. Making the enzyme an important indicator reflecting the relevant physiological characteristics in biological samples, accurate determination of pyruvate kinase activity in the kit is helpful for early detection and monitoring of the disease in medical diagnosis.

Components of buffer system

1 Tris buffer: Tris buffer is mainly used in the reagent kit to maintain a stable pH value of the reaction system. The activity of pyruvate kinase is highly sensitive to pH, and different pH conditions can cause significant changes in enzyme activity. Trimethylaminomethane can resist pH fluctuations caused by acidic or alkaline substances generated during the reaction process within a specific range. For example, during the reaction process, the consumption of substrates and the generation of products may change the acidity or alkalinity of the solution. Tris (hydroxymethyl) aminomethane buffer can maintain a relatively stable pH value through its own acid-base balance mechanism, ensuring that pyruvate kinase functions under suitable pH conditions.

2. The impact on other components: Tris (hydroxymethyl) aminomethane buffer also has a protective effect on other components in the reagent kit. It can prevent degradation or structural changes of substrates, coenzymes, and other components due to pH changes. At the same time, it has good compatibility with other reagents and will not undergo chemical reactions with phosphoenolpyruvate, ADP, NADH, etc., ensuring the stability and reliability of the entire reaction system.

Desheng Building

The various components in the pyruvate kinase potassium ion determination kit work together to achieve accurate determination of potassium ion concentration, from the reaction between substrate and coenzyme, to the stability of the buffer system and the guarantee of other auxiliary components. In the production, use, and quality control process of reagent kits, it is necessary to fully consider the characteristics and interrelationships of each component. The pyruvate kinase, Tris, and PEP produced by Hubei Xindesheng Material Technology Co., Ltd. can be used in potassium ion determination kits with high purity and stable performance feedback from cooperating kit manufacturers. If you have purchasing intentions, please feel free to click on the website for consultation at any time!