An improved method for assay of cholesterol 7α-hydroxylase activity
An improved method for assay of cholesterol 7α-hydroxylase activity
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DOI：
10.1016/0003-2697(86)90613-5
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发表时间：
1986-10
影响因子：
2.9
通讯作者：
T. Ogishima;K. Okuda
中科院分区：
文献类型：
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作者：
T. Ogishima;K. Okuda
描述了一种改进的胆固醇 7α-羟化酶测定方法，该方法准确、灵敏且简单。该方法由两部分组成：第一部分是以胆固醇为底物在肝微粒体内进行胆固醇7α-羟基化，第二部分是在胆固醇氧化酶的作用下将产物7α-羟基胆固醇转化为在240 nm处有强吸收的7α-羟基-4-胆固醇-3-酮。然后通过高效液相色谱分析转化的甾醇。在第二次酶反应期间，第一个酶反应停止并防止产物的进一步代谢。因此，该方法的灵敏度比之前的方法提高了 10 倍以上。
An improved assay method for cholesterol 7α-hydroxylase which is accurate, sensitive and yet still simple is described. The method consists of two parts: the first part is cholesterol 7α-hydroxylation in liver microsomes utilizing cholesterol in situ as the substrate, and the second part is conversion of the product, 7α-hydroxycholesterol, into 7α-hydroxy-4-cholesten-3-one having an intense absorption at 240 nm by the action of cholesterol oxidase. The converted sterol is then analyzed by high-performance liquid chromatography. During the second enzyme reaction, the first enzyme reaction is halted and further metabolism of the product is prevented. In consequence, the method had more than 10-fold increase in the sensitivity than the previous one.