中国林蛙皮肤抗菌肽的分离纯化及部分特性研究

Isolation of Skin Anti-bacteria Peptide of Chinese Forest Frog and Research of Its Partial Character

从长白山林蛙养殖场购买林蛙 ,用冷水清洗林蛙 ,断头后迅速剥离皮肤 ,清洗血液后剪碎 ,经 80 %的甲醇抽提 3次 ;再经组织捣碎机捣碎 ,用 80 %的甲醇抽提 2次 ;再经葡聚糖凝胶过滤。实验结果表明 ,林蛙皮肤中小分子活性多肽经葡聚糖凝胶过滤得到纯度较高的抗菌活性肽 ;林蛙皮肤中小分子活性多肽对革兰氏阳性细菌、革兰氏阴性细菌都具有一定的抗菌作用 ;经醋酸纤维薄膜电泳 ,表明其具有较强的阳离子特征 ;SDS - -PAGE电泳分析表明为单一区带 ,林蛙皮肤中小分子抗菌活性多肽的相对分子量为 6 .2 8ku。

The forest frog is bought from the forest frog plant of Changbai Mountain. The frogs are washed with cold water and the skin are peel off after decollation, and then cut to pieces after the blood was washed, extracted three times through methanol at concentration of 80%; then pounded to pieces by constitution triturator, extracted two times; passed sephadex gel. The result suggests that anti-bacteria bioactive peptide which has high purity can be gained by gel filtration from the small molecule bioactive peptide in the skin of forest frog, which has ability against gram-positive bacterium and gram-negative bacterium and has strong cation character by cellulose actate electeophoresis. SDS-PAGE analysis suggested that the small molecule bioactive peptide in the skin of forest frog is a single band and its molecular weight is 6.28 ku.