Harbron S, Eggelte H J, Rabin B R
London Biotechnology Ltd., Department of Biochemistry and Molecular Biology, University College London, England.
Anal Biochem. 1991 Oct;198(1):47-51. doi: 10.1016/0003-2697(91)90504-m.
Alkaline phosphatase hydrolyzes riboflavin 4'-phosphate to produce riboflavin. This is converted to riboflavin 5'-phosphate, using riboflavin kinase, which reconstitutes apoglycolate oxidase to give hologlycolate oxidase. This enzyme catalyzes the oxidation of glycolate with simultaneous production of hydrogen peroxide which is detected via the formation of a colored product through the action of peroxidase. The system allows the detection of 4 amol after a 2-h incubation.
碱性磷酸酶将核黄素4'-磷酸水解生成核黄素。利用核黄素激酶将其转化为核黄素5'-磷酸,该激酶可使脱辅基乙醇酸氧化酶重构为全乙醇酸氧化酶。这种酶催化乙醇酸的氧化,同时产生过氧化氢,过氧化氢通过过氧化物酶的作用形成有色产物来进行检测。该系统在孵育2小时后可检测到4阿摩尔。