粉末活性炭联合柱层析法纯化藻蓝蛋白的研究

Extraction and purification of c-phycocyanin from blue algae by combined use of powdered activated carbon treatment and column chromatography

以巢湖新鲜蓝藻藻泥为原料,通过冻融破壁的方法获得藻蓝蛋白粗提液,采用粉末活性炭法-羟基磷灰石柱层析法纯化藻蓝蛋白。在粉状活性炭法的研究中,通过单因素试验研究了粒径、添加量、藻蓝蛋白浓度和时间对藻蓝蛋白的提纯效果,并与羟基磷灰石柱层析法联用以优化高纯度藻蓝蛋白的提取纯化工艺组合。结果表明,粉末活性炭法纯化藻蓝蛋白的最优条件是粒径400~500目,藻蓝蛋白浓度1.0 mg·mL^-1,添加量100 g·L^-1和吸附时间15 min,经羟基磷灰石柱层析法纯化后,藻蓝蛋白纯度可达4.51,回收率为36%。

In this study,fresh cyanobacterial mud from Chaohu Lake was used as raw material,and algae was broke down by freezing and thawing to obtain crude phycocyanin.Then phycocyanin was purified by the combined use of powdered activated carbon adsorption and hydroxyapatite column chromatography.In the study of powdered activated carbon treatment,the effect of particle size,addition amount,phycocyanin concentration,and time on phycocyanin were studied by single-factor experiments,and then phycocyanin was purified by hydroxyapatite column chromatography to optimize the process combination of extraction and purification of high-purity phycocyanin.The results showed that the optimum conditions for purification of phycocyanin by powdered activated carbon treatment were mesh 400-500,phycocyanin concentration 1.0 mg·mL^-1,addition amount 100 g·L^-1 and time 15 min.After the purification of hydroxyapatite column chromatography,the purity and recovery rate of phycocyanin could reach to 4.51 and 36%,respectively.