鱼腥藻藻蓝蛋白的提取·分离纯化和抑菌研究

目的]对鱼腥藻藻蓝蛋白的提取进行优化,并对藻蓝蛋白粗品进行分离纯化、凝胶电泳以及抑菌作用研究。方法]采用反复冻融法提取藻蓝蛋白,液相色谱分离纯化藻蓝蛋白,SDS-PAGE电泳分析蛋白质成分,并利用96孔板法培养、酶标仪浓度检测,进行抑菌作用研究。结果]藻蓝蛋白提取的最佳条件为固液比1∶1 g/ml,冷冻时间60 min,硫酸铵饱和度为60%。藻蓝蛋白的最大吸收光是620nm。高效液相色谱分离得到3种蛋白质,标记为PC-1、PC-2、PC-3。SDS-PAGE电泳可知,PC-1和PC-2的分子量在14～18 kD。PC-2对苏云金芽孢杆菌和藤黄叠球菌有较弱的抑菌作用。结论]为鱼腥藻藻蓝蛋白的提取、纯化和性质研究提供了试验依据。

Study on the Extraction, Separation and Purification and Antibacterial Effect of Phycocyanin in Anabaena

Objective] The aim was to optimize the extraction of Anabaena phycocyanin,and study the separation and purification,gel electrophoresis and bacteriostatic effects on crude phycocyanin.Method] Using repeated freeze-thawing method to extract phycocyanin,and liquid chromatogram to separate and purify phycocyanin,SDS-PAGE electrophoresis to analyze protein component,moreover,using 96 orifice plate method culture and concentration measurement of enzyme standard instrument to study antibacterial effects.Result] The optimum extraction conditions of phycocyanin are: solid to liquid ratio of 1∶1,frozen time of 60 min,ammonium sulfate saturation of 60%.The maximum light-absorbing of phycocyanin is 620 nm.3 kinds of proteins are obtained based on high performance liquid chromatography and marked as PC-1,PC-2,PC-3,respectively.SDS-PAGE electrophoresis show that the molecular weights of PC-1 and PC-2 are among 14-18 kD.PC-2 has weaker antibacterial action on Bacillus thuringiensis and Micrococcus luteus.Conclusion] The study provided test basis for the extraction,purification and properties study of Anabaena phycocyanin.