钝顶螺旋藻中C-藻蓝蛋白的提取及性质

Extraction and characterization of C-phycocyanin from Spirulina platensis

摘要:

采用振荡提取法（Oscillatory extraction， OE）、高速匀浆提取法（High-speed homogenization extraction， HHE）、超声提取法（Ultrasonic extraction， UE）和反复冻融提取法（Repeated freeze-thaw extraction， RFE）4种技术手段从钝顶螺旋藻（Spirulina platensis）中分离制备C-藻蓝蛋白，并系统分析其分子结构特征、理化性质、功能特性及抗氧化生物活性效能。研究表明，4种提取方法所得C-藻蓝蛋白纯度都达到了试剂级标准，其中反复冻融提取法和高速匀浆提取法提取得到的C-藻蓝蛋白具有较高的纯度，分别达到了2.68和2.37。红外光谱分析显示，反复冻融得到的C-藻蓝蛋白α-螺旋含量最高，说明其二级结构最稳定。C-藻蓝蛋白的功能特性研究表明，其溶解度、乳化能力、起泡能力、持水性、持油性、最低凝胶浓度和表面疏水性均表现为反复冻融提取法>高速匀浆提取法>超声提取法>振荡提取法（P<0.05）。由SDS-PAGE图谱可知，所有提取方法得到的C-藻蓝蛋白均包含α 和β 两个螺旋亚基。抗氧化实验结果显示，C-藻蓝蛋白具有较强的抗氧化特性，4种提取方式所得C-藻蓝蛋白清除DPPH和ABTS自由基能力表现为反复冻融提取法>高速匀浆提取法>超声提取法>振荡提取法。综上所述，反复冻融提取法得到的C-藻蓝蛋白纯度最高，二级结构最稳定，具有最佳的功能特性，包括溶解度、乳化能力、起泡能力、持水性、持油性、最低凝胶浓度和表面疏水性，同时具有较强的抗氧化性，这些研究结果为拓展C-藻蓝蛋白在食品、医药和化妆品等领域的应用提供理论支持和研究基础。

Abstract:

The objective of this study is to extract C-phycocyanin from Spirulina platensis using four different extraction methods and to investigate its structural， physicochemical， functional properties， and antioxidant activity. The four extraction methods include oscillatory extraction （OE）， high-speed homogenate extraction （HHE）， ultrasonic extraction （UE）， and repeated freeze-thaw extraction （RFE）. The results showed that the purity of C-phycocyanin obtained by the four extraction methods reached the standard of reagent grade. The purity of C-phycocyanin obtained by repeated freeze-thawing and high-speed homogenization methods reached 2.68 and 2.37， respectively. FTIR analysis showed that C-phycocyanin α-helix obtained by repeated freezing and thawing had the highest content， indicating that its secondary structure was the most stable. The physicochemical and functional properties of C-phycocyanin showed that its solubility， emulsification ability， foaming ability， water holding capacity， oil holding capacity， minimum gel concentration and surface hydrophobicity all showed the following characteristics： repeated freeze-thaw extraction > high-speed homogenate extraction > ultrasonic extraction > oscillating extraction （P<0.05）. As shown by SDS-PAGE profiles， C-phycocyanin obtained by all extraction methods contained two helical subunits， α and β . The results of antioxidant experiments showed that C-phycocyanin had strong antioxidant properties. The ability of C-phycocyanin to scour DPPH and ABTS free radicals obtained by the four extraction methods was shown as repeated freeze-thaw extraction， high-speed homogenate extraction， ultrasonic extraction， oscillation extraction. In conclusion， C-phycocyanin extracted by repeated freezing and thawing has the highest purity， the most stable secondary structure， the best solubility， emulsification ability， foamability， water holding capacity， oil holding capacity， minimum gel concentration and surface hydrophobicity， and strong antioxidant ability. These results provide theoretical support and research basis for expanding the application of C-phycocyanin in food， medicine and cosmetics.