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Author:
Date:
2015-02-05
[Abstract] Intermediates of tetrapyrrole biosynthetic pathway are low-abundant compounds, and their quantification is usually difficult, time consuming and requires large amounts of input material. Here, we describe a method allowing fast and accurate quantification of almost all intermediates of the heme and chlorophyll biosynthesis, including mono-vinyl and di-vinyl forms of (proto) chlorophyllide, using just a few millilitres of the cyanobacterial culture. Extracted precursors are separated by High Performance Liquid Chromatography system (HPLC) and detected by two ultra-sensitive fluorescence detectors set to different wavelengths.
[摘要] 四吡咯生物合成途径的中间体是低丰度化合物，它们的定量通常是困难的，耗时的并且需要大量的输入材料。 在这里，我们描述了一种方法，允许快速和准确的量化几乎所有的血红素和叶绿素生物合成的中间体，包括单乙烯基和二乙烯基形式的（原）叶绿素，只使用几毫升的蓝藻培养。 提取的前体通过高效液相色谱系统（HPLC）分离，并通过设置为不同波长的两个超灵敏荧光检测器检测。
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