Q SepharoseFF分离藻红蛋白的工艺研究

摘要

[目的]建立QSepharoseFF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]QSepharoseFF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 SepharoseFF柱上，以1ml／min的流速使用添加0—0．10—0．35-1．00mol／LNaCl的NaH护O4-Na4-HPO4缓冲溶液（pH6．0）分步洗脱，收集0．35mol／L NaCl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20mlQSepharoseFF柱上进行分离，发现分离效果没有发生明显变化。[结论]使用QSepharoseFF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。%[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.