论文部分内容阅读
为了规模化制备具有抗肿瘤活性的海胆黄多糖(SEP),简化纯化步骤和提高多糖收率,在原有分离纯化方法的基础上,结合超滤技术,对SEP的纯化工艺进行了优化。采用木瓜蛋白酶-Sevag联用法,以超滤法替代减压浓缩和透析去除杂质,优化酶法提取条件和超滤参数。研究结果表明:用0.6%的木瓜蛋白酶在pH 6.5、60℃酶解5 h,可去除18.3%的蛋白质,3次Sevag法后,蛋白质去除率达71.7%,多糖回收率80.1%。采用截留相对分子质量100 k的膜超滤浓缩,压力0.05 MPa,原液质量浓度1.2 mg/mL,浓缩3倍时的多糖截留率为84.0%。各步骤多糖累计回收率为67.3%,该纯化工艺可获得高产率高纯度的SEP,为规模化制备SEP奠定了基础。
In order to prepare the SEP with anti-tumor activity, simplify the purification step and increase the yield of polysaccharide, the purification process of SEP was optimized based on the original separation and purification methods and ultrafiltration technology. Using papain-sevag combination, ultrafiltration instead of vacuum concentration and dialysis to remove impurities, optimize enzymatic extraction conditions and ultrafiltration parameters. The results showed that 18.3% protein could be removed by digestion with 0.6% papain at pH 6.5 and 60 ℃ for 5 h. After 3 times of Sevag method, the protein removal rate reached 71.7% and the polysaccharide recovery rate was 80.1%. The membrane was concentrated by membrane ultrafiltration with molecular weight cut-off of 100 kPa, the pressure was 0.05 MPa and the concentration of crude liquid was 1.2 mg / mL. The polysaccharide retention rate was 84.0% when the concentration was 3 times. The total recovery of polysaccharides in each step was 67.3%. This purification process can obtain SEP with high yield and high purity, which lays the foundation for the large-scale preparation of SEP.