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董宏坡, 左正宏, 陈奕欣. 坛紫菜中别藻蓝蛋白的纯化及洗脱组分光谱研究[J]. 植物科学学报, 2006, 24(6): 583-586.
引用本文: 董宏坡, 左正宏, 陈奕欣. 坛紫菜中别藻蓝蛋白的纯化及洗脱组分光谱研究[J]. 植物科学学报, 2006, 24(6): 583-586.
DONG Hong-Po, ZUO Zheng-Hong, CHEN Yi-Xin. Allophycocyanin Purification from Porphyra haitanensis and Studies on Spectrum of Eluting Fractions[J]. Plant Science Journal, 2006, 24(6): 583-586.
Citation: DONG Hong-Po, ZUO Zheng-Hong, CHEN Yi-Xin. Allophycocyanin Purification from Porphyra haitanensis and Studies on Spectrum of Eluting Fractions[J]. Plant Science Journal, 2006, 24(6): 583-586.

坛紫菜中别藻蓝蛋白的纯化及洗脱组分光谱研究

Allophycocyanin Purification from Porphyra haitanensis and Studies on Spectrum of Eluting Fractions

  • 摘要: 以福建省平潭县坛紫菜(Porphyra haitanensis)为材料,采用硫酸铵分级分离和柱层析法纯化别藻蓝蛋白(APC),并对纯化的条件进行了详细的探讨。研究结果表明:采用30%~35%饱和硫酸铵沉淀、Tris-HC l(pH=8.0)作为洗脱缓冲液、DEAE-Sephadex-A-50作为层析介质,所获得的APC的纯度和回收率分别为3.50和70.2%;SDS-PAGE表明APC有α和β两个亚基,分子量分别为18.9 kD和17.7 kD。因此该方法对于从坛紫菜中快速纯化APC是适合的。采用光谱分析研究柱洗脱组分,结果表明:坛紫菜中含有分子结构为(αβ)的“双峰型”R-PE,含有结构为(αβ)3的R-PC,含有结构为(αβ)3的APC-Ⅱ。

     

    Abstract: Allophycocyanin from Porphyra haitanensis collected in Pingtan County of Fujian Province was purified using ammonium sulfate precipitation and column chromatography.Meanwhile,factors of purification were studied in detail.The results showed that using 30%-35% of the saturation ammonium sulfate precipitation,Tris-HCl(pH=8.0)as eluting buffering and DEAE-Sephadex-A-50 as chromatographic adsorbent;the purity and rate of recovery of allophycocyanin obtained were 3.50 and 70.2%,(respectively).SDS-PAGE demonstrated the presence of two subunits,α and β,with Mr(18.9 kD) and(17.7 kD),respectively.Therefore the method was suitable for quick purification of allophycocyanin from P.haitanensis.At the same time,the fractions collected from column were researched by analysis of spectrum.The findings indicated that P.haitanensis contained the R-phycoerythrin with two absorption peaks which had the polypeptide composition(αβ),the R-phycocyanin which had the polypeptide composition(αβ)3 and allophycocyanin-II which had the polypeptide composition(αβ)3.

     

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