Purification, crystallization and preliminary X-ray analysis of phycocyanin and phycoerythrin from Porphyra yezoensis Ueda.
Purification, crystallization and preliminary X-ray analysis of phycocyanin and phycoerythrin from Porphyra yezoensis Ueda.
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DOI：
10.1107/s1744309111007925
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发表时间：
2011-05
期刊：
影响因子：
--
通讯作者：
Chuner Cai;Lian Wu;Chunxia Li;Peimin He;Jie Li;Jiahai Zhou
中科院分区：
文献类型：
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作者：
Chuner Cai;Lian Wu;Chunxia Li;Peimin He;Jie Li;Jiahai Zhou
条斑紫菜是中国最重要、最广泛养殖的海藻之一。藻胆蛋白具有优良的光谱特性，在生物医学、食品、化妆品和化学合成染料工业中发挥着广泛的作用。本文介绍了从条斑紫菜中提取的两种藻胆蛋白--藻红蛋白和藻蓝蛋白的纯化和结晶过程。利用硫酸铵共沉淀和羟基磷灰石柱层析的新工艺，这两种藻胆蛋白都得到了大规模生产，与以前报道的相比，质量和产量都有所提高。天然PAGE分析表明，藻红蛋白和藻蓝蛋白在溶液中以(αβ)(3)异六聚体形式存在。藻红蛋白晶体的衍射率为2.07？，属于R3空间群。参考六角轴的晶胞参数为a=b=187.7，c=59.7o，每个晶胞有9个(αβ)(2)异四异构体。在P2(1)空间群中，藻蓝蛋白晶体的衍射率达到2.70？分辨率。马修斯系数分析表明，藻蓝蛋白在不对称单元中的杂二聚体为10-19(αβ)是合理的。自旋函数的计算澄清了这种模糊性，并表明藻蓝蛋白的12个(αβ)杂二聚体组装在不对称单元中。
Porphyra yezoensis is one of the most important and widely cultured seaweeds in China. Phycobiliproteins exhibit excellent spectroscopic properties and play versatile roles in the biomedical, food, cosmetics and chemical synthetic dye industries. Here, the purification and crystallization of phycoerythrin and phycocyanin, two phycobiliproteins extracted from P. yezoensis, are described. Using a novel protocol including co-precipitation with ammonium sulfate and hydroxyapatite column chromatography, both phycobiliproteins were produced on a large scale with improved quality and yield compared with those previously reported. Native PAGE analysis indicated that phycoerythrin and phycocyanin exist as (αβ)(3) heterohexamers in solution. The crystals of phycoerythrin diffracted to 2.07 Å resolution and belonged to space group R3. The unit-cell parameters referred to hexagonal axes are a = b = 187.7, c = 59.7 Å, with nine (αβ)(2) heterotetramers per unit cell. The crystals of phycocyanin diffracted to 2.70 Å resolution in space group P2(1). Matthews coefficient analysis shows that 10-19 (αβ) heterodimers of phycocyanin in the asymmetric unit would be reasonable. A self-rotation function calculation clarified this ambiguity and indicated that 12 (αβ) heterodimers of phycocyanin are assembled in the asymmetric unit.