长白山林蛙输卵管糖蛋白ROGP-Ⅲ的分离纯化及鉴定

Separation and Identification of Oviduct Glycoprotein ROGP-Ⅲ from Rana chensinensis

通过正交试验确定最佳的粗提工艺,并将林蛙输卵管水提液先后通过盐析,透析,阴离子交换层析、阳离子交换层析和葡聚糖凝胶过滤层析,获得了一种分子量大小约为66 kD的单一蛋白,高碘酸-Schiff试剂法鉴定其为糖蛋白。经HPLC分析,纯度为97.2%,将其命名为ROGP-Ⅲ,经PNGase F糖苷肽酶处理后推测其糖含量约为17%。

To study the function of the single glycoprotein in Rana chensinensis, this research determined the best coarse extraction process by orthogonal test, then purified a 66 kD single protein by salting out, dialysis, ion-exchange column chromatography and gel filtration chromatography. The purified protein identified as glycoprotein by PAS test. By HPLC identification, its purity is 97.2%, named as ROGP-Ⅲ, after enzymolysis by PNGase F, it speculated that the polysaccharide content is about 17%.