海胆黄多糖SEP纯化工艺的研究

Isolation and Purification of the Polysaccharride from the Eggs of Strongylocentrotus Nudus(SEP)

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。

The polysaccharide from the eggs of Strongylocentrotus nudus（SEP） has significant anti-tumor activity. In order to isolate and purify SEP on a large scale, as well as simplify purification steps and improve the yield of polysaccharide, a practical extraction process from Strongylocentrotus Nudus eggs was investigated based on the original methods. Enzyme-Sevag method was used for deproteinization, and ultrafihration was applied to concentrate extraction solution and remove impurities. It is shown that 18.3% of protein was removed with 0.6% （w/w） of papain,under the condition of pH value 6. 5 at 60℃ for 5 h,and 71.7% of protein was removed from polysaccharide solution by deproteinizing with Sevag agent for three times with the polysaccharide recovery of 80.1%. The polysaccharide recovery rate reached 84.0% by uhrafihration （ MWCO 100 k,0.05 MPa, 1.2 mg/mL） with three times of concentration. Finally,the total recovery rate of polysaccharide was 67.3%. High yield and high purity of SEP was obtained through this process,which has laid the foundation for large-scale preparation of SEP.