全蝎胃蛋白酶酶解混合多肽体外抗凝活性研究及其组成分析

Fractions and anticoagulant activity in vitro of mixed polypeptide from pepsin hydrolysis of Quanxie(Scorpio)

目的采用凝胶过滤层析法分离全蝎酶解液的抗凝活性部位,并检测活性部分的分子量分布范围。方法采用紫外检测法、考马斯亮蓝法检测,合并凝胶层析蛋白流分;以APTT凝血时间为指标,检验各组分体外抗凝活性;通过MALDI-TOF-MASS法检测分离部分的分子量分布范围。结果全蝎酶解液通过凝胶过滤层析分得的2组分抗凝效果显著,分子量分布范围850~5 300。结论体外抗凝试验表明：凝胶过滤层析法纯化分离全蝎胃蛋白酶酶解液方法可行,MALDI-TOF-MASS表明活性组分为小分子肽类。

Objective To separate the fractions with anticoagulant activity from the pepsin hydrolytic liquid of Quanxie（Scorpio）,and detect the distribution of molecular weight in the active fractions by applying gel filtration chromatography.Methods The methods of UV-detection and Coomassie brilliant blue G250 were used to detect and combine protein fractions of gel chromatography.The anticoagulant activities of different fractions were detected taken activated partial thromboplastin time（APTT） as index.The distribution of molecular weight of the separated active fractions was determined by using MALDI-TOF-MS.Results There two fractions,separated from the pepsin hydrolytic liquid of Quanxie by gel filtration chromatography,with significant anticoagulant activities.The molecular weight distributed from 850 to 5 300.Conclusion The result of anticoagulant test in vitro indicates that gel filtration chromatography is feasible for purifying and separating the pepsin hydrolytic liquid of Quanxie.The outcomes of MALDI-TOF-MS show that the active fraction is micromolecular peptide.