麻疯树叶中过氧化物酶JCP-1的分离纯化和几种特性的检测  

Isolation,Purification and Analysis of Some Characters of Peroxidase from Leaves in Jatropha curcas L.

作　　者：叶生亮[1] 蔡峰[1] 朱勋路[1] 秦晓波[1] 徐莺[1] 陈放[1] 

机构地区：[1]四川大学生命科学学院,成都610064

出　　处：《植物生理学通讯》2010年第4期329-334,共6页Plant Physiology Communications

基　　金：国家自然科技资源平台项目(2005DKA21403);国家"十一五"科技支撑项目(2006BAD07A04)

摘　　要：麻疯树叶片蛋白粗提液经硫酸铵分级沉淀,强阴离子琼脂糖、强阳离子琼脂糖和交联葡聚糖层析,得到一个比活为4499U·mg-1(蛋白)过氧化物酶,命名为JCP-1。其分子量为49kDa,等电点为pH3.3,最适pH为5.0￣6.0。以H2O2为底物的Km为2.14mmol·L-1。JCP-1具有宽泛的最适保存pH(7.0￣11.0)和较高的耐热性(80℃高温处理15min,活性保持在90%以上)。30%PEG6000处理模拟干旱胁迫及50℃高温胁迫麻疯树苗,其叶片中JCP-1活性分别提高121%和155%。A peroxidase （JCP-1） was purified from leaves of Jatropha curcas using a multi-step process includ- ing ammonium sulfate fractionation, Q Sepharose F F, SP sepharose F F and Superdex 75 Prepgrade chromatography. The purified enzyme exhibited specific activity of 4 499 U·mg-1 （protein）. The molecular weight examined by SDS-PAGE was 49 kDa. The isoelectric point was pH 3.3. The optimum pH value of JCP-1 was 5.0–6.0. The results of kinetic study showed its high affinity to H2O2 with Km value of 2.14 mmol·L-1. JCP-1 showed a high thermal stability that it retained over 90% of the activity after 15 min at 80 ℃. The activity was also stable at a broad pH range of 7.0–11.0. JCP-1 activity in the leaves of J. curcas increased significantly under 30% PEG6000 or 50 ℃.

关 键 词：麻疯树 过氧化物酶 纯化 酶学性质 稳定性 抗逆性 

分 类 号：Q946[生物学—植物学]