摘要
为了降低天然大分子功能蛋白的分离纯化成本,以琼脂为原料自制弱阴离子交换层析介质DEAE-琼脂凝胶微球。将DEAE-琼脂凝胶微球应用于坛紫菜R-藻红蛋白(R-PE)和螺旋藻C-藻蓝蛋白(C-PC)的提取分离,最佳洗脱条件为:层析柱规格h=35.0 cm,φ=1.0 cm,V=27.5 m L;流速v=0.33 m L/min;洗脱缓冲液为0.01 mol/L磷酸盐溶液(含有0.05~0.5 mol/L Na Cl溶液);0.01 mol/L磷酸盐缓冲液(含0.1 mol/L Na Cl)洗脱出纯度为5.0的坛紫菜R-PE;0.01 mol/L磷酸盐缓冲液(含0.2 mol/L Na Cl)洗脱出纯度为4.4的螺旋藻C-PC。自制层析介质成本低廉、耗能少、可大量制备、分离效果好,可用于坛紫菜R-PE、螺旋藻C-PC的产业化制备。
In order to reduce the separation and purification cost of natural macromolecular functional protein,a kind of weak anion exchange chromatography medium DEAE-agar gel microsphere was made up. Applied DEAE-agar gel microsphere to the separation and purification of the porphyra haitanensis R-phycoerythrin( R- PE) and spirulina C-phycocyanin( C- PC), optimal elution conditions were as follows : chromatography column specifications was h=35.0 cm,φ=1.0 cm,V=27.5 m L,current velocity v=0.33 m L/min,elution buffer solution was 0.01 mol/L phosphate buffer solution(contain 0.05~0.5 mol/L Na Cl). 0.01 mol/L phosphate buffer solution(contain 0.1 mol/L Na Cl) washed porphyra haitanensis R-PE out,its purity was 5.0. 0.01 mol/L phosphate buffer solution(contain 0.2 mol/L Na Cl) washed spirulina C-PC out,its purity was 4.4. The DEAE-agar gel microspheres could be used in the industrial production of R-PE and C-PC because of their low cost,low energy consumption,scale preparation and excellent separation effect.
引文
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