Oizumi J, Hayakawa K
National Children's Medical Research Center, Division of Metabolism, Tokyo, Japan.
J Chromatogr. 1990 Jul 13;529(1):55-63. doi: 10.1016/s0378-4347(00)83807-4.
A convenient and reliable reversed-phase liquid chromatographic assay for four kinds of carboxylase, i.e. pyruvate carboxylase, acetyl-coenzyme A carboxylase, propionyl-coenzyme A carboxylase, and 3-methylcrotonyl-coenzyme A carboxylase, was developed. These are all biotin enzymes, which contain covalently bound biotin as a co-factor (co-enzyme). This chromatographic method, which is free from interferences, gives a stoichiometric and precise assay for these enzymes. It is non-radioisotopic, which makes it suitable for clinical laboratories. The assay time required for one sample is ca. 30 min. The method was satisfactorily applied to guinea-pig kidney homogenates after a simple pretreatment with acid.
建立了一种简便可靠的反相液相色谱法，用于测定四种羧化酶，即丙酮酸羧化酶、乙酰辅酶A羧化酶、丙酰辅酶A羧化酶和3-甲基巴豆酰辅酶A羧化酶。这些都是生物素酶，它们含有共价结合的生物素作为辅因子（辅酶）。这种色谱方法不受干扰，能对这些酶进行化学计量和精确测定。它是非放射性同位素的，这使其适用于临床实验室。测定一个样品所需的时间约为30分钟。该方法经简单酸预处理后，成功应用于豚鼠肾脏匀浆。
