Purification and characterization of phycoferritin from the blue-green alga, Arthrospira (Spirulina) platensis
Purification and characterization of phycoferritin from the blue-green alga, Arthrospira (Spirulina) platensis
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蓝绿藻钝顶节旋藻（螺旋藻）中藻铁蛋白的纯化和表征
DOI：
10.1007/s10811-007-9263-7
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发表时间：
2008
影响因子：
3.3
通讯作者：
V. Deshpande
中科院分区：
文献类型：
--
作者：
G. Lakshmi Deepa;R. Sashidhar;V. Deshpande
应用常规生化技术，从营养价值很高的蓝藻--钝顶螺旋藻中分离出藻铁蛋白。纯化蛋白的相对分子质量为470kDa，产率为0.044 mg g−1，铁含量为1.4%，总中性糖含量为20%。与细菌和哺乳动物的铁蛋白相比，铁的含量要低得多。藻铁蛋白的P：Fe比为1：3.5，与细菌铁蛋白相近。天然凝胶电泳法显示有异构体存在。经SDS-PAGE和Western blotting亚基分析，发现一个表观分子量为18 kDa的蛋白质亚基。蛋白质亚单位的低聚形式也存在。藻铁蛋白与抗豌豆种子铁蛋白发生交叉反应，提示与高等植物的系统发育关系。GC-MS分析藻铁蛋白的碳水化合物发现，藻铁蛋白中含有类似哺乳动物铁蛋白的半乳糖、葡萄糖和甘露糖等糖。有趣的是，分析还发现了鼠李糖、木糖和塔尔糖等糖，这些糖在铁质的结构中还没有报道。除藻铁蛋白中的组氨酸含量很低外，其余氨基酸组成与其他物种的铁蛋白相似。藻铁蛋白的紫外可见光谱分析表明，存在血红素基，这是细菌铁蛋白的特性。藻铁蛋白的荧光强度高于马脾铁蛋白。圆二色谱显示出较低的螺旋度。
Phycoferritin from the nutritionally important blue-green alga Arthrospira platensis has been isolated, by application of conventional biochemical techniques. The molecular mass, yield, iron and total neutral carbohydrate contents of the purified protein were 470 kDa, 0.044 mg g−1 of Arthrospira, 1.4 and 20%, respectively. The iron content was much lower when compared to bacterial and mammalian ferritins. The P: Fe ratio of Arthrospira phycoferritin was 1: 3.5, a value akin to bacterioferritins. Native gel-electrophoresis revealed the presence of isoforms. Subunit analysis by SDS-PAGE and Western blotting showed a protein subunit with an apparent molecular mass of 18 kDa. Oligomeric forms of the protein subunit were also present. The phycoferritin exhibited cross-reactivity with anti-pea seed ferritin suggesting phylogenetic relationship with that of higher plants. Carbohydrate analysis of phycoferritin by GC-MS revealed the presence of sugars such as galactose, glucose and mannose similar to that of mammalian ferritins. Interestingly, the analysis also revealed sugars such as rhamnose, xylose and talose, which has not been reported in the structure of ferritins. Except for very low histidine content in phycoferritin, the rest of the amino acid composition resembled to ferritins of other species. UV-visible spectral analysis of the phycoferritin revealed the presence of haem groups, a property characteristic of bacterioferritins. The fluorescence intensity of phycoferritin was higher than equine spleen ferritin. Circular dichroic spectra revealed a lower degree of helicity.
影响因子：
7.1
作者：
Murray-Kolb, LE;Welch, R;Beard, JL
通讯作者：
Beard, JL