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Author:
Date:
2014-08-05
[Abstract] Riboflavin is the precursor of flavin nucleotides FMN and FAD, they play significant roles in all organisms. GTP is the initial precursor on riboflavin biosynthesis pathway and GTP cyclohydrolase II catalyzes the first step of this pathway. It converts GTP to 2,5-diamino-6-ribosylamino-4 (3H) -pyrimidinone 5'-phosphate. This protocol provides a reliable and fast method to assay GTP cyclohydrolase II activity from crude bacterial extracts. The product of the reaction catalyzed by GTP cyclohydrolase II, 2,5-diamino-6-ribosylamino-4 (3H) -pyrimidinone 5'-phosphate, is converted to its fluorescent derivative 6,7-dimethylpterin, which is then separated on a XTerra MS C18 column and detected using fluorescence HPLC system.
[摘要] 核黄素是黄素核苷酸FMN和FAD的前体，它们在所有生物体中起重要作用。 GTP是核黄素生物合成途径的最初前体，GTP环化水解酶II催化该途径的第一步。 它将GTP转化为2,5-二氨基-6-核糖基氨基-4（3 H） - 嘧啶酮5'-磷酸。 该方案提供了从粗细菌提取物中测定GTP环化水解酶II活性的可靠且快速的方法。 由GTP环化水解酶II 2,5-二氨基-6-核糖基氨基-4（3 H） - 嘧啶酮5'-磷酸催化的反应产物转化为其荧光衍生物6,7- 二甲基蝶呤，然后在XTerra MS C18柱上分离，并使用荧光HPLC系统检测。
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