【摘要】 采用反相高效液相法(RP-HPLC)测定磷酸果糖激酶的活性。将磷酸果糖激酶与含有果糖-6-磷酸和ATP的反应体系混合,置于30℃水浴中,反应20 min后,沸水浴3 min终止反应。通过RP-HPLC测定酶反应前后产物二磷酸腺苷(ADP)量的变化来分析酶的活性。ADP的生成量在5~400 mg/L范围内具有良好的线性关系,方法的精密度良好(RSD<5%)。本方法与通常所用的酶偶联法(ECM)相比准确度高,且简便易行。更多还原

【Abstract】 Phosphofructokinase(PFK,EC2.7.1.11) activity was determined by reversed-phase high performance liquid chromatography(RP-PHLC) in this paper.After PFK,Fructo-6-phospho and ATP were mixed in the reaction system,the mixture was incubated at 30℃ for 30 min,and the reaction was stopped in the boiling water for 3 min.The quantity of product ADP before and after reaction which was determined by RP-PHLC was dependent on the PFK activity.The product ADP had good linear relationship at 5~400 mg/L with a good precision(RSD<5%).RP-PHLC was more accurat than the conventional enzyme coupling method(ECM),and its operation was more convenient.更多还原