花芸豆铁蛋白的分离纯化

Isolation and Purification of Ferritin from the Phaseolus vulgaris

为了从花芸豆中获取纯化的植物铁蛋白,试验利用盐析及柱层析对花芸豆中的铁蛋白进行分离纯化。研究表明,铁蛋白提取的最佳工艺条件为:提取温度53.79℃,样液p H值为7.23,氯化镁浓度511.15mmol/L,柠檬酸三钠浓度为685.24 mmol/L。该条件下模型预测花芸豆中铁蛋白的含量为4.40 mg/g,实际试验值为(4.27±0.09)mg/g。经DEAE-Sepharose Fast Flow阴离子交换柱和Sephacryl S-500分子筛柱纯化得到的电泳纯铁蛋白的相对分子质量为560 000,且只含有1种亚基,其相对分子质量为27 000。通过Western-Blot法对纯化所得的花芸豆蛋白质进行试验验证,结果确定该蛋白质为铁蛋白。

In order to obtain the purified phytoferritin from the Phaseolus vulgaris, ferritin was isolated and purified through salting out method and column chromatography. As a result, the ferritin yield was up to （4.27 ± 0.09 ） mg/g under the condition that extraction temperature 53.79 ℃, sample solutions pH 7.23, concentration of MgC12 511.15 mmol/L, concentration of sodium citrate 685.24 mmol/L. Ferritin of electrophoresis level, the molecular weight of 560 000, was obtained by further chromatographic purification which contains only one subunit of 27 000. Simultaneously, protein extracted from the phaseolus vulgaris was identified with the help of the Western Blot. The result revealed that the protein was ferritin.