正北芪中一种免疫活性蛋白质提取工艺的优选

陈秀红; 任晋宏; 魏砚明; 薛慧清; 武福云; 李敏; 梁锐

doi:10.13422/j.cnki.syfjx.2016050013

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正北芪中一种免疫活性蛋白质提取工艺的优选

药剂与炮制 | 更新时间：2024-01-04

- 正北芪中一种免疫活性蛋白质提取工艺的优选
- Optimization of Extraction Technology of An Immune Active Protein from Astragali Radix
- 中国实验方剂学杂志 2016年22卷第5期页码：13-17
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家国际科技合作专项(2013DFA30700)
- DOI：10.13422/j.cnki.syfjx.2016050013
  中图分类号： R284.2
- 纸质出版日期：2016
- 稿件说明：
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陈秀红, 任晋宏, 魏砚明, 等. 正北芪中一种免疫活性蛋白质提取工艺的优选[J]. 中国实验方剂学杂志, 2016,22(5):13-17.

CHEN Xiu-hong, REN Jin-hong, WEI Yan-ming, et al. Optimization of Extraction Technology of An Immune Active Protein from Astragali Radix[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(5): 13-17.
陈秀红, 任晋宏, 魏砚明, 等. 正北芪中一种免疫活性蛋白质提取工艺的优选[J]. 中国实验方剂学杂志, 2016,22(5):13-17. DOI： 10.13422/j.cnki.syfjx.2016050013.

CHEN Xiu-hong, REN Jin-hong, WEI Yan-ming, et al. Optimization of Extraction Technology of An Immune Active Protein from Astragali Radix[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(5): 13-17. DOI： 10.13422/j.cnki.syfjx.2016050013.

摘要

目的: 优选正北芪中一种免疫活性蛋白AmPR10-16(16.0 kDa)的提取工艺. 方法: 以正北芪中AmPR10-16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AmPR10-16在SDS-PAGE胶图中的蛋白条带吸光度(峰面积)为指标

采用L9(34)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AmPR10-16提取工艺的影响

辅以BCA法测定可溶性蛋白含量作为佐证. 结论: AmPR10-16最佳提取工艺为药材粉末过4号筛

加16倍量pH 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.AmPR10-16提取率0.063 g ·g-1. 结论: 优化的提取工艺能正确反映AmPR10-16提取率的最高相对量

为AmPR10-16的进一步研究提供了稳定、合理、可行的提取工艺.

Abstract

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.

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