Study on Separation of Phycoerythrin by Q-Sepharose Fast Flow  

Q Sepharose FF分离藻红蛋白的工艺研究(英文)

作　　者：张彬[1,2] 王翠芹[3] 高敏[3] 张轶[4] 顾铭[3] 王长海[1] 

机构地区：[1]中国科学院烟台海岸带研究所,山东烟台264003 [2]中国科学院研究生院,北京100049 [3]中国科学院过程工程研究所,北京100190 [4]南京农业大学,江苏南京210095

出　　处：《Agricultural Science & Technology》2012年第8期1641-1644,共4页农业科学与技术（英文版）

基　　金：Supported by National Natural Science Foundation of China(51143012);Natural Science Foundation of Shandong Province(ZR2009BM006)~~

摘　　要：[Objective] This study aimed to establish an efficient process for separation of phycoerythrin by using Q Sepharose Fast Flow resin and verity its feasibility for scale-up. [Method] Elution gradient, sample volume and flow rate were optimized to determine the optimal separation condition, under which the scale-up process was verified. [Result] The optimal condition for separation of phycoerythrin by using Q Sepharose FF resin was investigated： 30 ml of laver extract was loaded to the Q Sepharose FF column with a bed volume of 8 ml; subsequently, the column was stepwise eluted with 0-0.10-0.35-1.00 mol/L NaCI solution （pH 6.0） at a constant flow rate of 1 ml/min; the elution peak under 0.35 mol/L NaCI solution was collected, and the recovery rate and purity coefficient （A565/A280） of phycoerythrin were determined as 44.3 and 1.15, respectively. Based on the established process, 75 ml of phycoerythrin extract was loaded to the Q Sepharose FF column with a bed volume of 20 ml for separation, while no significant variation was observed in the separation result. [Conclusion] Phycoerythrin can be well separated from laver extract by using Q Sepharose FF resin and the process is feasible for scale-up.[目的]建立Q Sepharose FF分离藻红蛋白的工艺方法并验证其放大可行性。[方法]通过对洗脱条件、上样体积以及洗脱流速的优化,确定最佳分离工艺条件;在确定的工艺条件下,将上样体积与柱体积等比例放大。[结果]Q Sepharose FF分离藻红蛋白的最佳工艺条件为:将30ml坛紫菜提取液加载到8ml Q Sepharose FF柱上,以1ml/min的流速使用添加0-0.10-0.35-1.00mol/LNaCl的NaH2PO4-Na2HPO4缓冲溶液(pH6.0)分步洗脱,收集0.35mol/LNaCl溶液洗脱时的色谱峰。在些条件下,R-藻红蛋白的回收率和纯度系数(A565/A280)分别为44.3和1.15。按照该工艺,将75ml藻红蛋白提取液加载到20ml Q Sepharose FF柱上进行分离,发现分离效果没有发生明显变化。[结论]使用Q Sepharose FF介质能够分离坛紫菜提取液中的藻红蛋白且该工艺易于放大。

关 键 词：Ion exchange chromatography Q Sepharose Fast Flow PHYCOERYTHRIN SEPARATION Scale-up 

分 类 号：TQ936.2[化学工程]