苯扎贝特降解酶活力检测方法的优化实验研究

Experimental Study on Optimal Detection Method for Bezafibrate Degradation Enzyme Activity

为进一步了解菌株PseudomonasputidaB一31对苯扎贝特的降解机制，对其降解酶活力的检测方法进行．『实验研究。优化确定了苯扎贝特降解酶的超声破碎提取方法，探讨了反应温度、pH值、反应时间以及酶浓度对苯扎贝特降解酶活力的影响。结果表明，苯扎贝特降解酶的最佳提取条件为：超声功率150W，运行时间20min，工作时间3S，休息时间2S，降解酶活力对温度和pH值变化较为敏感，最佳测定条件为：pH=7．0，反应温度30℃，反应时间2h，酶浓度80～100mg／L，反应时间2h。提取的降解酶与苯扎贝特亲和力较好，其米氏常数‰和最大反应速度％分别为41．85μmol／L和0．074μmol／（L·min）。

Assay optimization of degradation enzyme detection was studied for further understanding the degradation mechanism for bezafibrate of Pseudomonas putMa B-31 strain. Optimum ultrasonication extraction conditions were determined and effect of reaction temperature, pH value, reaction time and enzyme concentration on bezafibrate degradation enzyme activity was investigated. Results showed that the optimum conditions for enzyme extraction were ultrasonic power of 150 W, running time of 20 rain, working time of 3 s and resting time of 2 s. The extracted enzyme reactivity was sensitive to the variation of reaction temperature and pH value, and the optimum detection conditions were pH value 7.0,reaction temperature 30 ~C, reaction time 2 h and enzyme concentration 80~100 mg/L. Additionally, the extracted enzyme was found to be of a favorable affinity to bezafibrate, with Michaelis constant and the maximum reaction rate as 41.85 μmol/L and 0.074 μmol/（L.min） respectively.