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双水相萃取是近年来发展起来的蛋白纯化方法。本研究运用双水相技术分离提取钝顶螺旋藻中藻蓝蛋白,对构成双水相体系中不同分子量PEG和酒石酸钾钠浓度的影响进行了分析。确定了双水相组成体系为16%的PEG 2000(w/w)和25%的酒石酸钾钠(w/w),pH值为6.0,在此体系中藻蓝蛋白主要分布在上相,最高纯度3.69,分配系数20.7,回收率94.56%。多次双水相萃取有利于藻蓝蛋白纯度提高,3次双水相萃取后,藻蓝蛋白纯度高达4.15。
Double aqueous extraction is a protein purification method developed in recent years. In this study, the separation and extraction of phycocyanin from Spirulina platensis using aqueous two-phase technology was studied. The effects of different molecular weight PEG and potassium sodium tartrate on the aqueous two-phase system were analyzed. The aqueous two-phase system was determined to be PEG 2000 (w / w) at 16% and potassium sodium tartrate (w / w) at 25% at pH 6.0. Purity 3.69, partition coefficient 20.7, recovery 94.56%. Multiple aqueous two-phase extraction is conducive to the increase of the purity of phycocyanin. After three times of aqueous two-phase extraction, the purity of phycocyanin reaches 4.15.