荧光法测定蛇毒中磷脂酶的活性

An ASSAY OF PHOSPHOLIPASE FROM SNAKE VENOM BY FLUORESCENCE

利用二醋酸荧光素(FDA)可被磷脂酶水解,而本过程受卵磷脂抑制这一原理.对蛇毒及其它样品中的磷脂酶活性进行测量.初步探讨了该法的实验条件及其机理.研究结果表明,这个新方法可以简便、特异地测量出样品中的磷脂酶活性.我们利用该法测定了广东眼镜蛇粗毒及分离样、浙江蝮蛇粗毒,得到了和前人工作相近的结论;而在牛血清白蛋白中可检出磷脂酶活性,植物血凝素以及伴刀豆球蛋白中则检测不出磷脂酶活性.

A new method has been described for determination of phospholipase activities in snake venom, which is based on the fact that fluorescein diacetate (FDA) is enzymatically hydrolyzed by phospholipase and the hydrolysis is inhibited after the addition of lecithin. The primary exploration on the optimal enzymati-cal condition and the machanism of this method were carried out. The results demonstrated that the present method is reliable, specific and convenient for the assay of phospholipase. The activities of phospholipase from crude and isolated venom of Naja naja Atra, and crude venom of Agkistrodon halys Pallas was respecti velly investigated, and the data were consistent with previous works. BSA has phospholipase activities, and PHA, ConA has no at all.