人参蛋白抗衰老机制研究

项目名称： 人参蛋白抗衰老机制研究

项目编号： No.30873371

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 赵雨

作者单位： 长春中医药大学

项目金额： 30万元

中文摘要： 采用膜分离技术提取人参总蛋白，建立了一条快速、简单、高效的人参总蛋白纯化工艺路线，较传统硫酸铵沉淀法，更适合于工业化生产。 从细胞及动物整体水平证明了人参总蛋白的抗衰老活性。人参蛋白对L929细胞增殖具有明显的促进作用；动物实验表明具有明显的抗氧化、抗疲劳、耐缺氧功效。 针对过氧化物酶（POD）、苹果酸脱氢酶（MDH）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）等4种氧化还原酶，进行了不同品种、不同部位及不同生长年限的活性对比研究。不同部位人参各种酶活力均有较大差异，不同年限人参各种酶活力虽有一定差异，但差别不大。 从人参中分离纯化了具有清除自由基活性的人参超氧化物歧化酶（SOD），并对其性质进行了研究。与其他植物来源SOD相比，人参SOD具有活力高，热稳定性及pH稳定性较好的优点。成功构建人参pBV220-SOD温控表达载体，实现了人参SOD基因在大肠杆菌中以包涵体形式表达，通过纯化表达蛋白得到电泳纯的人参SOD蛋白，纯化产物酶活较高。为人参SOD的开发应用奠定基础。

中文关键词： 人参；蛋白；抗衰老

英文摘要： Using a membrane filtration purification system, we had established a fast, simple and highly effective method to purify the total protein from Ginseng, which should be more appropriate for industrial scale protein production, We also confirmed the anti-aging activity of our Ginseng total protein in cell assay and in-vivo animal experiment. Our total ginseng protein show significant stimulation to L929 cell proliferation. Meanwhile, the animal experiment indicate a clear anti-oxidation, anti-fatigue and hypoxia tolerance effect. The activities of four redoxenzyme, i.e., peroxidase (POD), superoxide dismutase(SOD), malate dehydrogenase (MDH) and catalase (CAT), in total ginseng protein of different varieties, positions and lifetimes, were compared. We found significant difference in the samples from different position, whereas no significant difference was found for that of different lifetimes. We further purified the SOD from the total protein extract and characterized it. The purified ginseng SOD possess certain advantages such as, higher activity, better thermo and pH stability,etc. Thereafter, we constructed a temperature-control expression system using pBV-220 vector carrying ginseng SOD gene sequence, to heterologously express the ginseng SOD in E. Coli as inclusion body. Purification from this strategy yield high activity, single band SOD protein on gel electrophoresis. The presentstudy provides a solid foundation further studies and application of ginseng SOD.

英文关键词： Ginseng;protein;anti-aging