海滨锦葵籽粒蛋白的分离、纯化及其生物活性测定  

Separation,Purification and Bioactivities Determination of Kosteletzkya virginica Seed Protein

作　　者：张焕仕[1] 李思宇[1] 谢银[1] 张鹤云[1] 张太平[1] 钦佩[1] 

机构地区：[1]南京大学盐生植物实验室,南京210093

出　　处：《天然产物研究与开发》2013年第12期1685-1689,共5页Natural Product Research and Development

基　　金：江苏省产学研联合创新资金项目(BY2012136)

摘　　要：以海滨锦葵籽粒脱脂粉为原料,采用超声波法、DEAE-纤维素柱层析、浓度梯度洗脱模式(洗脱液盐浓度分别为0.1、0.25、0.4 mol/L)分离纯化得到三个蛋白组分;再对0.1 mol/L盐洗脱蛋白进行超滤分离。采用SDS-PAGE测定纯化蛋白分子量的分布,并利用MTT法测定蛋白的体外免疫和抗肿瘤活性。结果表明,总水溶性蛋白得率为1.12%;0.25 mol/L盐洗脱冻干粉的蛋白质量分数最高为58.8%;含量较高的蛋白质分子量分别为23、32和60KD;0.1 mol/L盐洗脱蛋白表现出较好的促进脾淋巴细胞增殖和抑制HepS细胞增殖的作用。随着时间延长,小于30KD的超滤蛋白组分对HepS细胞生长表现出显著或极显著抑制效果。Degrease powder of Kosteletzkya virginica seeds were used as raw materials in this research. Three components of protein were purifiedby ultrasound, DEAE-ceUulose column chromatography with gradient concentration desorption- mode （ the concentrationsof desorption saltwere 0.1,0.25 and 0.4 mol/L,respectively）. 0.1 mol/L salt desorption com- ponents were further separated using ultrafiltration. Molecular weight distribution of three protein components were deter- mined by SDS-PAGE. In vitro immunization and antitumor activities of proteins were tested by M＇IT assay. The results showed that the total soluble protein extraction rate was 1.12% ;the highest protein mass percent was 58.8% from 0.25 mol/L salt desorption concentration;with higher levels of protein molecular weight 23,32 and 60KD. 0.1 mol/L salt de- sorption protein （SDP） showed significant effects on promoting proliferation of splenic lymphocyte and inhibiting prolif- eration of mouse hepatoma cell （HepS）. Components of smaller than 30KD showed significant and very significant inhib- itory effects on proliferation of HepS cells with the increasing of time.

关 键 词：海滨锦葵 水溶性蛋白 分离 纯化 生物活性 

分 类 号：R284[医药卫生—中药学] R73-36[医药卫生—中医学]