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葡萄糖-6-磷酸脱氢酶是磷酸戊糖途径的关键限速酶,催化葡萄糖-6-磷酸的氧化反应。 其活性检测基于经典的NADPH生成速率法。G6PD催化葡萄糖-6-磷酸氧化生成6-磷酸葡萄糖酸-δ-内酯,并同时将氧化型辅酶Ⅱ还原为还原型辅酶Ⅱ。在340 nm波长下,NADPH具有特征吸收峰,其生成速率与反应体系中G6PD的活性成正比。因此,通过监测340 nm处吸光度随时间上升的速率,即可准确计算出样本中G6PD的酶活性。
Glucose-6-phosphate dehydrogenase is the key rate-limiting enzyme in the pentose phosphate pathway, catalyzing the oxidation of glucose-6-phosphate. Its activity assay is based on the classical NADPH generation rate method. G6PD catalyzes the oxidation of glucose-6-phosphate to produce 6-phosphogluconate-δ-lactone, while simultaneously reducing oxidized coenzyme II (NADP⁺) to reduced coenzyme II (NADPH). NADPH exhibits a characteristic absorption peak at a wavelength of 340 nm, and its generation rate is directly proportional to the G6PD activity in the reaction system. Therefore, by monitoring the rate of increase in absorbance at 340 nm over time, the G6PD enzyme activity in the sample can be accurately calculated.