大鼠肝脏和纤细裸藻中苯丙氨酸羟化酶活性测定不同方法的比较。
Comparison of different methods for the determination of phenylalanine hydroxylase activity in rat liver and Euglena gracilis.
作者信息
Fink R M, Elstner E F
出版信息
Z Naturforsch C Biosci. 1984 Jul-Aug;39(7-8):728-33. doi: 10.1515/znc-1984-7-808.
Three different methods for the determination of phenylalanine hydroxylase activity have been compared: a) Differential photometric assay of the increase in tyrosine concentration in the presence of phenylalanine; b) Product separation by thin layer chromatography and scintillation counting of the [14C]tyrosine formed; c) HPLC separation and spectrofluorometric quantification of derivatized amino acids. A comparison of the activities of phenylalanine hydroxylase in rat liver and Euglena gracilis clearly showed that only rat liver contains this enzymic activity as shown by methods b) and c) although pseudo-activity of Euglena gracilis preparations was found during the spectrophotometric test a). The HPLC method proved to be the fastest, most reliable and convenient method for direct tyrosine determination and thus for measuring phenylalanine hydroxylase activity.
已比较了三种测定苯丙氨酸羟化酶活性的不同方法
a）在苯丙氨酸存在下，通过差示光度法测定酪氨酸浓度的增加；b）通过薄层色谱法分离产物，并对形成的[14C]酪氨酸进行闪烁计数；c）通过高效液相色谱法分离并对衍生化氨基酸进行荧光光谱定量。大鼠肝脏和纤细裸藻中苯丙氨酸羟化酶活性的比较清楚地表明，只有大鼠肝脏含有这种酶活性，如方法b）和c）所示，尽管在分光光度法测试a）中发现纤细裸藻制剂存在假活性。高效液相色谱法被证明是直接测定酪氨酸从而测量苯丙氨酸羟化酶活性的最快、最可靠且最方便的方法。
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