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Author:
Date:
2014-08-05
[Abstract] This protocol provides a simple and fast method of quantification for intracellular flavin content, and for flavin secretion by bacteria. Intracellular flavins are extracted from bacterial pellets, and secreted flavins are examined in the cell growth medium. Flavins are separated and measured using HPLC with fluorescence detection, and quantified based on a comparison to standards.
[摘要] 这个协议提供了一种简单和快速的方法定量细胞内黄素含量，和黄素分泌的细菌。 从细菌团块提取细胞内黄素，并在细胞生长培养基中检查分泌的黄素。 分离黄酮并使用具有荧光检测的HPLC测量，并基于与标准品的比较进行定量。
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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 ...
[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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