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作 者:李冰[1] 张学成[2] 高美华[1] 褚现明[3]
机构地区:[1]青岛大学医学院,山东青岛266012 [2]中国海洋大学海洋生命学院,山东青岛266003 [3]青岛市海慈医院,山东青岛266033
出 处:《海洋科学》2007年第8期48-52,共5页Marine Sciences
摘 要:对钝顶螺旋藻(Spirulina platensis)中藻蓝蛋白的提取和纯化方法进行了改进。用磷酸盐缓冲液循环冻融联合超声波破碎法,50%硫酸铵沉淀获得藻蓝蛋白(phycocyanin,PC),提取率达到13.1%。粗蛋白提取液再经过两次羟基磷灰石柱(HA)层析和sephacryl-HR-200凝胶层析对其进行纯化,纯度达到4.71%。纯化后的PC在12%十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)中得到两条带,分别是α和β两个亚基,分子质量分别为21.4 ku和17.0 ku。结果表明,通过上述分离纯化过程得到了较高提取率和电泳纯度的藻蓝蛋白。The optimum method of extraction and purification of phycocyanin from Spirulina platensis was improved in this paper. Through repeatedly freezing and thawing alternatively in the presence of phosphate buffer combined with ultrasonic fragmentation, the phycocyanin was efficiently extracted from S. platensis by being precipitated in 50% solid ammonium sulphated, resulting in 13.1% recovery of phycocyanin. Then, the crude phycocyanin was chromatographed by hydroxylapatite (HA) column, further purified by sephacrylHR- 200 gel chromatography and HA column chromatography again. In the end the phycocyanin with a high purity (A620/A280) of 4.71 was obtained. Analyzed by 12% SDS-PAGE electrophoresis, phycocyanin migrated as two bands corresponding to its two subunits (α and β) and the molecular weights were 17.0 ku and 21.4 ku, respectively. The results showed that we got the phycocyanin with a comparatively high extracting rate and a high purity by means of a series of extraction and purification procedures.
关 键 词:钝顶螺旋藻(Spirulina platensis) 藻蓝蛋白 提取 纯化
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