木薯叶过氧化物酶的分离纯化及其同工酶的研究  

Separation and Purification of Peroxidase from Cassava Leaves and Study on Its Isozyme

作　　者：陈燕珍[1,2] 谭涛[1,2] 杨栋林[1,2] 刘洁雯[1] 李谋岚[1] 

机构地区：[1]广西师范大学生命科学学院,广西桂林541004 [2]广西环境工程与保护评价重点实验室,广西桂林541004

出　　处：《安徽农业科学》2009年第36期17872-17873,17909,共3页Journal of Anhui Agricultural Sciences

基　　金：广西环境工程与保护评价重点实验室研究基金项目(桂科能0702k025)

摘　　要：[目的]探讨将木薯叶作为过氧化物酶（POD）制备原料的可行性。[方法]木薯叶和甘蔗叶经提取液提取、盐析法沉淀,得POD粗酶液,分别用Sephadex G-75层析法和切向流超滤法纯化POD,用聚丙烯酰胺凝胶电泳（PAGE）法分离了POD同工酶。[结果]木薯叶和甘蔗叶POD的酶活分别为6.617×10~4和5.883×10~3U/g,前者远高于后者。SephadexG-75层析法分离POD粗酶液得到洗脱峰Ⅰ和Ⅱ,木薯叶和甘蔗叶都是峰Ⅰ有酶活,峰Ⅱ酶活很低甚至没有。POD粗酶液经切向流超滤后,木薯叶和甘蔗叶POD的回收率分别为91.825%和90.820%,RZ值分别为2.75和2.85。约在430nm处有最大吸收峰。木薯叶POD有3条与标样辣根POD相同的同工酶带。[结论]木薯叶POD含量高,提取纯化操作简便,有开发利用价值。[Objectives]To investigate the feasibility that cassava leaves are used as the raw materials of peroxidase（POD）.[Methods]POD crude enzyme solutions were obtained by extracting cassava leaves and sugar cane leaves and,in turn,salting - out precipitation.Purification of POD was performed by using Sephadex G-75 gel chromatography and tangential flow ultrafiltration,respectively.POD isozymes were separated by using polyacrylamide gel electrophoresis（PAGE）.[Results]The POD activities of cassava leaves and sugar cane leaves are 6.617×10~4 and 5.883×10~3 U/g,respectively,and the former is much higher than the latter.PeakⅠand peakⅡwere obtained after purifying POD crude enzyme solutions by using Sephadex G-75 gel chromatography.PeakⅠhas POD activity in cassava leaves and sugar cane leaves;however, peakⅡhas very low or even no POD activity.Tangential flow ultrafiltration showed that the recovery rates of POD activity of cassava leaves and sugar cane leaves are 91.825%and 90.820%,respectively,RZ value is 2.75 and 2.85,respectively,and maximum absorption peak appears at about 430 nm.The POD isozymes of cassava leaf have 3 electrophoresis bands,which is the same as standard sample of horseradish POD.[Conclusions]Cassava leaves have high content of POD,and the methods of extraction and purification are simple and convenient, thus,cassava leaves can be used for exploiting POD.

关 键 词：木薯叶 甘蔗叶 过氧化物酶 超滤 电泳 

分 类 号：S533[农业科学—作物学]