Author:
Date:
2014-05-05
[Abstract] A recent study demonstrated that cytosolic phosphoenolpyruvate carboxykinase (PEPCK) and NADP-malic enzyme (NADP-ME) have an important role in malate metabolism during fruit ripening (Osorio et al., 2013). PEPCK catalyze the ATP-dependent decarboxylation of oxaloacetate (OAA) to phosphoenolpyruvate (PEP) and NADP-ME, the reversible conversion of malate and pyruvate. Here, we present the detailed protocols to measure PEPCK activity in carboxylation direction by following oxidation of NADH and to measure NADP-ME activity based upon the reduction of NADP+.
[摘要] 最近的研究表明胞质磷酸烯醇丙酮酸羧激酶(PEPCK)和NADP-苹果酸酶(NADP-ME)在果实成熟期间在苹果酸代谢中具有重要作用(Osorio等人,2013)。 PEPCK催化草酰乙酸(OAA)向磷酸烯醇丙酮酸(PEP)和NADP-ME的ATP依赖性脱羧,苹果酸和丙酮酸的可逆转化。 在这里,我们提出详细的协议,以测量PEPCK活性的羧化方向通过以下氧化NADH和基于NADP + 的还原测量NADP-ME活性。
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