用于测定粗生物提取物中吡哆醇激酶活性的简单等度高效液相色谱法。
Simple isocratic high-performance liquid chromatographic method for measuring pyridoxine kinase activity in crude biological extracts.
作者信息
Argoudelis C J
机构信息
Department of Food Science, University of Illinois, Urbana 61801.
出版信息
J Chromatogr. 1990 Mar 16;526(1):25-33. doi: 10.1016/s0378-4347(00)82480-9.
To measure the enzymatic activity of pyridoxine kinase (EC 2.7.1.35) when pyridoxine or pyridoxamine are the substrates an additional enzymatic step is usually required. The products of the kinase activity, pyridoxine 5'-phosphate or pyridoxamine 5'-phosphate, are oxidized, enzymatically, to pyridoxal 5'-phosphate (co-enzyme) which is then measured either spectrophotometrically or using apo-enzymes. In this report the enzymatic activity of pyridoxine kinase, in crude biological extracts, is assayed by a simple high-performance liquid chromatographic method which determines the amount of pyridoxine 5'-phosphate formed when pyridoxine is the substrate. The same method could be used when pyridoxamine is the substrate.
当以吡哆醇或吡哆胺作为底物来测定吡哆醇激酶（EC 2.7.1.35）的酶活性时，通常还需要额外的酶促步骤。激酶活性的产物，即5'-磷酸吡哆醇或5'-磷酸吡哆胺，会被酶促氧化为5'-磷酸吡哆醛（辅酶），然后通过分光光度法或使用脱辅基酶来进行测定。在本报告中，粗生物提取物中吡哆醇激酶的酶活性通过一种简单的高效液相色谱法进行测定，该方法可测定以吡哆醇作为底物时形成的5'-磷酸吡哆醇的量。当以吡哆胺作为底物时，也可使用相同的方法。
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