阴离子交换柱(DEAE-52)层析纯化海参自溶酶

Purification of holothurians autoenzyme using chromatography on DEAE-52 cellulose

本研究针对海参自溶酶的特点,经过实验确立了以下纯化方案:首先将海参湿肠打浆,再用0.1mol/L,pH6.3磷酸盐缓冲溶液浸提,离心弃沉淀得上清液;然后将乙醇加入到上清液中,使其浓度达30%,离心取沉淀。最后采用阴离子交换柱(DEAE 52)层析,得到在SDS 聚丙烯酰胺电泳(SDS PAGE)上表现为一条带的海参自溶酶样品。

In accordance with the characteristics of the holothurians' autoenzyme, an effective separating and purifying plan is determined via experiment as follows: crushing the holothurians gut is crushed, extracting the protease with phosphate buffer(0.1 mol/L,pH 6.3) and retaining the liquid by discarding the sediments, adding ethanolto the precipitated liquid with a concentration of 30%, centrifuging and precipitating, chromatographying on DEAE-cellulose. The sample obtained is a single protein band on SDS-PAGE(Sodium dodecyl sulfate-polyalkylene glycol ether).