Osborne法分级提取五味子蛋白及抗氧化活性比较

Extraction of Schisandra chinensis Protein by Osborne Method and Its Antioxidant Activity

目的:采用Osborne法提取五味子各组分蛋白,并对其体外抗氧化活性比较研究。方法:用Osborne方法分级提取五味子中的蛋白质,得到四种组分蛋白:清蛋白、球蛋白、醇溶蛋白和谷蛋白,对其进行十二烷基硫酸钠-聚丙烯酰胺凝胶(sodium lauryl sulfate-polyacrylamide gel,SDS-PAGE)电泳分析,并对不同组分蛋白和总蛋白的体外抗氧化活性进行对比研究。结果:分级提取得到的四种蛋白质,清蛋白亚基分子量分布在15~25 kDa和40~55 kDa,球蛋白分子量大致分布在15~25 kDa和35~55 kDa,谷蛋白亚基分子量在15~20 kDa和30~40 kDa。以五味子总蛋白为参照物,体外抗氧化试验结果表明,总蛋白对羟基自由基和DPPH自由基的清除能力强于各分级蛋白;四种分级蛋白对Fe3+的还原能力强于总蛋白;在ABTS自由基清除试验中含量最高的谷蛋白效果最佳,其清除效果在一定的浓度下与Vc接近,同时,其在羟基、DPPH自由基与Fe3+还原能力的试验中抗氧化活性良好,表明五味子组分蛋白中谷蛋白可作为一种良好的抗氧化潜在物质。

Objective:To extract the protein of Schisandra chinensis by Osborne method and compare its antioxidant activity in vitro. Methods:Osborne method classification was used to extract the protein in fructus schisandrae, get four components of protein: Albumin, globulin, gliadin and protein, sodium dodecyl sulfate-polyacrylamide gel to(sodium lauryl sulfate,polyacrylamide gel, SDS-PAGE) electrophoresis analysis, and the different components of protein and total protein in vitro antioxidant activity. Results: The four proteins obtained by Osborne method. The molecular weights of albumin subunits were 15~25 kDa and 40~55 kDa, the molecular weights of globulin were 15~25 kDa and 35~55 kDa, and the molecular weights of gluten subunits were 15~20 kDa and 30~40 kDa. The results of antioxidant test in vitro showed that the total protein of Schisandra chinensis had stronger scavenging ability on hydroxyl radical and DPPH radical than that of the different fractions of protein. The reduction ability of the four graded proteins to Fe3+ was stronger than that of the total proteins. In ABTS free radical scavenging test, the highest content of gluten had the best scavenging effect, which was close to Vc at a certain concentration. At the same time, the antioxidant activity of gluten in hydroxyl, DPPH free radical and Fe3+reducing ability test showed that gluten in Schisandrae chinensis component protein could be a good antioxidant potential substance.