木瓜蛋白酶在染料Cibacron Blue F3GA纤维素亲和膜上的吸附研究

以纤维素滤纸膜为载体,染料Cibacron Blue F3GA为配基,制备了一种新型亲和膜色谱介质。采用扫描电镜、红外光谱、元素分析等方法对亲和膜介质进行鉴定与表征,该膜具有良好的色谱性能。亲和膜对F3GA的键合质量摩尔浓度达93.7μmol/g。研究了木瓜蛋白酶在亲和膜上的吸附行为,实验表明:在30℃下、酶质量浓度为2 mg/mL、pH=8.0时,吸附质量比可达57.9 mg/g,改变pH值及离子强度等条件对吸附质量比有明显的影响。在最适条件下吸附遵循Langmuir型吸附。可以初步推断,纤维素滤纸膜可以制成性能优良的亲和膜色谱介质,成本低廉,适合工业化分离纯化生物大分子。

Adsorption of papain with Cibacron Blue F3GA carrying cellulose affinity membranes

Cibacron Blue F3GA (CB F3GA) as a hgand was immobilized onto cellulose membranes to produce a novel affinity membrane. The physical properties and its apphcations of affinity membrane chromatography were examined by means of scanning electron microscope (SEM), infra-red spectrum and elementary analysis, etc. The bonding content of CB F3GA attached on membranes was 93.7 μmol/g. The adsorption behavior of papain on affinity membranes was studied. The result shows that higher papain adsorption capacity (up to 57.9 mg/g membrane) can be achieved under the condition of 2.0 mg/mL papain solution, 30℃, pH=8.0. Changing pH and ionic strength has obvious effects on the adsorption of papain. The adsorption of papain on affinity membranes can be described by the Langmuir isotherm. Therefore, it can prehminarily foresee that the cellulose membrane can become the low-cost but high-efficiency affinity membranes base for papain separation, which is applicable for commercial separating the biological macromolecular.