摘要
为利用太湖打捞蓝藻的蛋白质资源,采用冻融破壁、絮凝、盐析、透析、双水相萃取等方法制备荧光试剂级藻蓝蛋白。研究结果表明,以水为介质冻融2次破壁,将冻融藻浆中蓝藻干物质调节为0.95%,用浓度为7.5g/L的聚合氯化铝絮凝初分离,所得藻蓝蛋白纯度(A6 20/A280)为0.31;采用20%、50%饱和度的硫酸铵两步盐析可将其纯度(A620/A280)提高至2.29;盐析后藻蓝蛋白经100 000的透析袋透析,再以140 g/L的聚乙二醇(分子量2 000)和140 g/L的磷酸钾盐萃取纯化,最终纯度(A620/A280)高达4.60,达到荧光试剂级要求。
To utilize the protein resources in blue algae in Lake Taihu, an approach, which included repeated freeze-thaw cycles, floceulation, ammonium sulfate salting-out, and aqueous two-phase extraction was developed to obtain high purity phycocyanin. By freezing and thawing blue algae two times in water, the concentration of algae dry matter was adjusted to 0. 95%. After flocculation by 7.5 g/L polymeric aluminum chloride, the phycoeyanin with the purity （A620/ A280 ） of 0. 31 was isolated. The purity was improved to 2. 29 through two-step salting-out with ammonium sulfate. Following the salting-out, the phycocyanin was dialyzed in a dialysis bag with molecular weight cut off of 100 000, and was then ex- tracted with 140 g/L polyethylene glycol 2000 and 140 g/L potassium phosphate salt. Those steps led to a higher purity of phyeocyanin, （A620/A280） being 4. 60, which met the standard of fluorescent reagent grade.
出处
《江苏农业学报》
CSCD
北大核心
2012年第4期777-782,共6页
Jiangsu Journal of Agricultural Sciences
基金
江苏省太湖水环境治理科研项目(TH2010205)
关键词
蓝藻
藻蓝蛋白
提取
纯化
blue algae
phycocyanin
extraction
purification
