中国传统发酵食品中溶栓酶活性检测方法比较

通过正交实验设计建立最佳琼脂糖-纤维蛋白平板法,并比较改进的纤维蛋白平板法和四肽底物法测定中国传统发酵食品中溶栓酶的活力的线性和精密度,以找出一个灵敏易操作、经济实惠、快速准确地标示溶栓酶效价的活性检测方法。实验结果显示纤维蛋白平板法和四肽底物法分别在50～800IU.mL-1、0.03～2.60U.mL-1范围内具有良好的线性关系,且日间和日内变异系数均小于6%。因此,纤维蛋白平板法与四肽底物法均能用于溶栓酶的活性检测,前者测定结果比较直观,但四肽底物法操作比较简单,经济成本相对较低,灵敏度高,更适合溶栓菌株的高通量筛选。

Comparison between Methods of Detecting Thrombolytic Enzyme Activity in Chinese Traditional Fermented Food

To seek a convenient,fast and accurate method for detecting fibrinolytic activity in Chinese traditional fermented food,the improved fibrin plate method and tetra-peptide substrate method were evaluated and compared between the linearity and the precision,and their results’ correspondence was also investigated.The results showed that the activity linear ranges were 50~800 IU·mL-1 for fibrin plate method and 0.03~2.60 U·mL-1 for tetra-peptide substrate method respectively,while both CVs were less than 6% in within-day and day-to-day tests.Therefore,the fibrin plate method and the tetra-peptide substrate method are both applicable for detecting fibrinolytic activity in Chinese traditional fermented food.The fibrin plate method can directly reflect fibrinolytic activity in Chinese traditional fermented food.The tetra-peptide substrate method was a convenient,economic,fast,sensitive and suitable approach for high-throughput screening of bacterial strains in Chinese traditional fermented food showing thrombolytic activity.