摘要
目的:优选正北芪中一种免疫活性蛋白Am PR10-16(16.0 k Da)的提取工艺。方法:以正北芪中Am PR10-16的可溶性蛋白二级结构的圆二色性考察提取温度,在单因素试验基础上,以Am PR10-16在SDS-PAGE胶图中的蛋白条带吸光度(峰面积)为指标,采用L_9(3~4)正交试验考察提取温度、料液比、提取时间、提取溶剂、粒度、提取次数对Am PR10-16提取工艺的影响,辅以BCA法测定可溶性蛋白含量作为佐证。结果:Am PR10-16最佳提取工艺为药材粉末过4号筛,加16倍量p H 8.0的三羟甲基氨基甲烷盐酸盐(Tris-HCl)于40℃常压水浴提取60 min,每10 min搅拌1次。Am PR10-16提取率0.063 g·g^(-1)。结论:优化的提取工艺能正确反映Am PR10-16提取率的最高相对量,为Am PR10-16的进一步研究提供了稳定、合理、可行的提取工艺。
Objective： To optimize extraction technology of an immune active protein （AmPR10-16） with a molecular weight of 16.0 kDa from Astragali Radix. Method： Extraction temperature was investigated by circular dichroism of water-soluble protein involving in AmPR10-16 with secondary structure from Astragali Radix.Extraction technology was optimized by single factor test and orthogonal test with gray value of AmPR10-16 as index which was determined by gel graphical analysis software. In this study,effects of temperature,solid-liquid ratio,time,solvent,granularity and times on gray value were investigated,for which determination of water-soluble protein was determined as an evidence by BCA method. Result： Optimum extraction process of AmPR10-16 was as follows：with Tris-HCl with pH 8.0 as solvent,solid-liquid ratio of 1 ： 16,extraction time of 60 min with temperature at 40 ℃,medicinal powder over the 4th sieve,stirring once every 10 min.Yield of AmPR10-16 was 0.063 g · g^-1. Conclusion： This optimal extraction technology can accurately reflect relative amounts of AmPR10-16 maximum extraction rate,it provides a stable,reasonable and feasible extraction process for furter study of AmPR10-16.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第5期13-17,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家国际科技合作专项(2013DFA30700)
