摘要
【目的】研究太子参的最佳蛋白提取工艺和不同参形太子参蛋白条带差异。【方法】选取3种蛋白提取方法(Tris-HCl法、TCA-丙酮法、饱和酚法)对贵州施秉县牛大场太子参种植基地的3种参形太子参进行蛋白提取及SDSPAGE分析,利用响应面分析法对筛选出的最佳蛋白提取方法的提取工艺进行优化。【结果】3种蛋白提取方法中Tris-HCl法提取的蛋白含量较高,经SDS-PAGE分析,太子参蛋白条带大致分布在10 k Da^90 k Da之间,15 k Da处一等参的蛋白表达量明显高于二等参和统货。利用响应面优化法获得提取太子参蛋白的最佳工艺。【结论】获得太子参蛋白的最佳提取工艺:料液比1∶1.32、提取温度25℃、pH值7.47、提取时间1.5 h,还发现不同参形太子参蛋白在15 k Da处的表达量存在明显差异,这可为今后利用蛋白质组学技术进一步揭示不同参形太子参形成的分子机制提供理论依据。
【Objective】This experiment was to study the protein extraction methods and protein bands differences of different parameters from Pseudostellariae Radix.【Method】Three kinds of protein extraction methods(Tris-HCl method,TCA-acetone method and Saturated phenol method)were used to extract proteions of three different parameters of Pseudostellariae Radix from the plantation area of Niudachang in Shibing,Guizhou.The extracted proteins were analyzed by SDS-PAGE electrophoresis.Response surface analysis was used to optimize the extraction process of the best protein extraction method.【Result】The content of protein extracted by Tris-HCl was highest in the three protein extraction methods.SDS-PAGE analysis showed that the protein bands were distributed between 10 kDa and 90 kDa in Pseudostellariae Radix,and the protein expression of first class at 15 kDa was significantly higher than that of the other two grades.The optimum process for extracting the Pseudostellariae Radix protein was obtained by response surface optimization.【Conclusion】The best extraction technology of Pseudostellariae Radix proteins were as follow:the material liquid ratio 1:1.32,extraction temperature 25℃,pH 7.47,extraction time 1.5 h.It was also found that there was a significant difference in the expression level of Pseudostellariae Radix proteins at 15 kDa in different parameters,which could provide a theoretical basis for further revealing the molecular mechanism of the formation for different parameters in Pseudostellariae Radix with proteomics technology in the future.
作者
晋海军
王海霞
张涛
向守艳
JIN Hai-jun;WANG Hai-xia;ZHANG Tao;XIANG Shou-Yan(Guiyang University of Chinese Medicine,Guiyang 550025,China)
出处
《四川农业大学学报》
CSCD
北大核心
2018年第4期500-506,共7页
Journal of Sichuan Agricultural University
基金
贵州省国内一流学科建设项目(GNYL(2017)008)
贵州省科技计划项目(黔科合LH字[2016]7117)
关键词
太子参
参形
SDS-PAGE分析
响应面优化
Pseudostellariae Radix
parameter
SDS-PAGE analysis
response surface optimization
