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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
磷酸烯醇式丙酮酸羧激酶（PEPCK）属于裂解酶类，根据核苷酸辅因子依赖性分为两种类型：一类是ATP依赖性PEPCK（ATP-PEPCK，EC 4.1.1.49），主要存在于开花植物、藻类及部分真菌和细菌中；另一类是GTP依赖性PEPCK（GTP-PEPCK，EC 4.1.1.32），主要存在于哺乳动物、鸟类、鱼类、昆虫、软体动物、扁虫、线虫、眼虫及部分真菌和细菌中。本试剂盒专一检测ATP依赖性PEPCK的活性，采用偶联酶动力学法连续监测其催化效率：PEPCK催化草酰乙酸（OAA）与ATP缩合生成磷酸烯醇式丙酮酸（PEP）、ADP和CO₂；生成的PEP在丙酮酸激酶（PK）催化下，将ADP磷酸化为ATP，同时自身转化为丙酮酸；丙酮酸在乳酸脱氢酶（LDH）催化下，被NADH还原为乳酸，同时NADH氧化为NAD⁺。通过监测340 nm处NADH特征吸收峰的下降速率，在底物饱和条件下，单位时间内OD₃₄₀的初始线性降幅（ΔOD₃₄₀/min）与PEPCK酶活性呈正比，据此即可精准计算样品中ATP-PEPCK的酶活力大小。
Phosphoenolpyruvate carboxykinase (PEPCK) belongs to the lyase family and is classified into two types based on nucleotide cofactor dependence: ATP-dependent PEPCK (ATP-PEPCK, EC 4.1.1.49), which is primarily found in flowering plants, algae, and some fungi and bacteria; and GTP-dependent PEPCK (GTP-PEPCK, EC 4.1.1.32), which is mainly present in mammals, birds, fish, insects, mollusks, flatworms, nematodes, euglenoids, and some fungi and bacteria. This kit specifically measures the activity of ATP-dependent PEPCK using a coupled enzymatic kinetic method for continuous monitoring of its catalytic efficiency: PEPCK catalyzes the condensation of oxaloacetate (OAA) and ATP to generate phosphoenolpyruvate (PEP), ADP, and CO₂. The generated PEP, under the catalysis of pyruvate kinase (PK), phosphorylates ADP to ATP while being converted to pyruvate. Pyruvate is then reduced to lactate by NADH under the catalysis of lactate dehydrogenase (LDH), with NADH being oxidized to NAD⁺. By monitoring the rate of decrease in the characteristic absorbance peak of NADH at 340 nm, under substrate-saturated conditions, the initial linear decrease in OD₃₄₀ per unit time (ΔOD₃₄₀/min) is proportional to PEPCK activity. Accordingly, the ATP-PEPCK enzyme activity in the sample can be precisely calculated.
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