文档介绍：摘要
目的:人参蛋白及多肽主要具有抗脂质分解、抗血红细胞聚积、抗真菌、抗病毒、精氨酸酶活性等。选择研究较少但活性多样的大分子-人参蛋白及多肽,通过现代分离纯化技术、活性检测技术及分析鉴定技术进行生物特性研究。通过对不同参之间的蛋白成分进行比较和研究,对人参的一些多肽成分的分析和应用提供依据。
方法:采用中药化学及生物化学提取法,结合离子交换、凝胶过滤层析和反相C18层析等现代生物分离技术, 利用膜分离技术及柱层析技术对总蛋白进行分离纯化,结合细胞活性实验进行活性筛选,对人参中的蛋白及多肽进行了分离纯化,采用HPLC、SDS-聚丙烯酰胺凝胶电泳法(SDS-PAGE)及凝胶电泳图像分析软件等谱学技术,初步研究了这些蛋白多肤的结构,并从细胞水平进行了体外活性实验,探讨蛋白与多肤的生物活性。
结果:采用现代生物层析分离技术从人参中分离纯化得到3个蛋白质单体化合物,分别为GSPⅠ、GSPⅡ及GSPⅢ。经基质辅助激光解吸电离飞行时间质谱人参蛋白及多肽药理活性实验表明, 人参蛋白GSPll对金黄色葡萄球菌具有抑制作用;对 Hep-2 细胞具有抑制作用,对3T3 细胞具有增殖作用,且活性呈现一定量效关系。
关键词人参多肽提取工艺活性研究
ABSTRACT
Objective:Ginseng proteins and polypeptide, which have a variety of biological activities but less reasearches, were studied by modern technologies including isolation and purification technology, activity detection technology and analysis - identification technology. Providing a basis for identification and quality evaluation among ginseng, wild ginseng and American ginseng by showing parative study of their water-soluble ponents.
Methods: Using different buffer, different ratio of solution, different extraction time to extract the ginseng protein, using Bradford method, determin the rate of protein extraction for the evaluation Standards, choose the best optimized extraction process. Purification was carried by hollow fiber system and column chromatography technology, combined with cell activity to screening ponent. Compare the difference of the proteins among ginseng, wild ginseng and American ginsengbyusing SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and image analysis software.
Results: Three proteins were obtained from Ginseng:GSPⅠ、GSPⅡand GSPⅢ。activity experiments show that it have inhibitory effect on 3T3 cells proliferation and have inhibitory effect on Hep-2 cells proliferation,
Key words Ginseng Polypeptide Extraction ProcessPropert
目录
第一章文献综述 1
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第二章材料与仪器 6
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第三章实验方法 8
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