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以南海海域的部分小杂鱼肝胰脏为原料,经磷酸盐缓冲液抽提,硫酸铵分级沉淀,SephadexG-100凝胶柱层析纯化来制备马尾藻多糖降解酶。系统的分析了硫酸铵饱和度、pH、盐析温度以及盐析时间对盐析效果的影响。结果表明:当硫酸铵饱和度为80%、盐析pH为5.5、盐析温度为45℃、盐析时间为3h时盐析效果最好。经SephadexG-100柱层析纯化后比活力达到126.72U/mg,提纯倍数达8.45倍,回收率为32.26%。
Heterophan hepatopancreas was isolated from the liver and pancreas of part of the South China Sea using phosphate buffered saline, ammonium sulfate fractionation and SephadexG-100 gel column chromatography. The effects of ammonium sulfate saturation, pH, salting-out temperature and salting-out time on salting-out effect were systematically analyzed. The results show that the salting out effect is the best when the saturation of ammonium sulfate is 80%, the salting-out pH is 5.5, the salting-out temperature is 45 ℃ and the salting-out time is 3h. After purification by SephadexG-100 column, the specific activity reached 126.72U / mg, the purification fold reached 8.45 times and the recovery rate was 32.26%.