大蒜SOD酶的分离纯化和肝素修饰  

Separation and Purification of SOD from Allium sativum L.and Its Chemical Modification by Heparin

作　　者：杨华[1] 周小苗[1] 方亚东[1] 郭纯[2] 龚银[1] 曹杏芝[1] 

机构地区：[1]湖南农业大学,湖南长沙410128 [2]湖南中医药大学药学院,湖南长沙410007

出　　处：《湖南农业科学》2011年第7期115-117,共3页Hunan Agricultural Sciences

基　　金：湖南省大学生研究性学习与创新性实验计划项目(2009413);湖南农业大学东方科技学院大学生创新项目(DFCXS200901)

摘　　要：采用热变性法、盐析法与有机溶剂提取法进行分离纯化,建立了一套适合大蒜SOD酶分离纯化的工艺流程:大蒜SOD酶提取液经70℃加热20 min,取上清在4℃条件下先使硫酸铵的饱和度达到40%;除杂后调整硫酸铵饱和度达到80%,取沉淀透析;透析液经纤维素层析,NaCl醋酸缓冲液梯度洗脱,收集第3个蛋白峰。针对天然SOD酶的不稳定性,采用了生物大分子肝素结合修饰进行化学修饰,结果表明:肝素修饰法有效提高SOD酶溶液的保存期。By using heat denaturation method,salting-out method and organic solvent extraction method,a set of process flow suited separation and purification of SOD from Allium sativum L.was established as follows： after heat extract solution of SOD at 70℃ for 20 min,the supernatant was taken to make saturation of ammonium sulfate to be 40% at 4℃;then the saturation of ammonium sulfate was adjusted to 80% after removing impurities,and the sediments were taken to dialyze;and then,the dialysate was through cellulose chromatography and gradient elution with NaCl buffer solution,and the 3rd protein peak was collected.According to the instability of natural SOD,the chemical modification was conducted to the SOD that combining biomacromolecule heparin with modification,the results indicated that heparin modification can effectively prolong the preservation period of SOD solution.

关 键 词：SOD 分离纯化 纤维素层析 肝素 化学修饰 

分 类 号：Q946[生物学—植物学]