明绿豆苯丙氨酸解氨酶的分离纯化及性质初步研究

Purification and Properties of Phenylalanine Ammonia- Lyase in Ming Mung Bean

为了研究明绿豆中苯丙氨酸解氨酶（PAL）的动力学特性，采用35％和65％饱和度的硫酸铵分级沉淀、DE-52阴离子交换层析方法对明绿豆中的PAL进行了分离纯化，并对其基本性质做了初步研究。结果表明，PAL的纯化倍数为6．9829，蛋白质得率为4．65％，酶得率为32．45％。PAL在硼砂-硼酸缓冲溶液中适宜的pH范围为7．6～9．0，最适pH有2个：8．0和8．6。PAL适宜的温度范围为35—45℃，最适反应温度为40℃。得到PAL的2个Km值，分别是：Km1为6.94×10^-5 mol/L,Km2为1.23×10^-4 mol/L。本研究结果可为进一步研究和开发明绿豆的PAL提供参考。

The study in this paper aimed to research the dynamics characteristics of phenylalanine ammonia - lyase （PAL） from Ming mung bean. The isolation, purification and basic properties of L - phenylalanine ammonia - lyase in Ming mung bean were studied by 35% and 65% saturation of ammonium sulfate grade deposition, DE -52 anion exchange chromatography. The results showed that the purification multiple was 6. 982 9 and protein yield and PAL yield were 4.65% and 32.45% respectively. The optimal pH range of PAL in borax - boric acid buffer was 7.6 - 9.0 and there were two optimal pH as 8.0 and 8.6. The optimal temperature range of PAL was 35 - 45 ℃. The optimal temperature was 40 ℃. There were two Km values including 6.94 × 10 ^-5 mol/L and 1.23 × 10 ^-4 mol/L. The results provided the reference for further studying and developing PAL of Ming mung bean.