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我们利用VY/4平板法和酪蛋白平板法从水样、土样和朽木样品中筛选出6株可产生抗血栓活性物质的可滑动的细菌菌株,重点研究了菌株B4和B8,利用萃取法及(NH4)2SO4沉淀法对菌株B4和B8的胞外活性蛋白进行了初步分离。一方面,我们利用纤维蛋白平板法及加热的纤维蛋白平板法检测到菌株B4和B8的胞外蛋白可以通过直接降解纤维蛋白的方式溶解血栓,利用纤维蛋白原平板法及凝血酶平板法检测到菌株B4和B8的胞外蛋白可以通过降解纤维蛋白原的方式防止血栓的形成;另一方面,我们利用形态学特征、理化性质、16S r RNA序列同源性及系统进化树鉴定菌株B4和B8均属于苏云金芽孢杆菌(Bacillus thuringiensis)。综上所述,苏云金芽孢杆菌B4和B8的胞外蛋白既具有溶栓活性,又具有抗凝活性,即具有抗血栓的作用。
We used VY / 4 plate method and casein plate method to screen out 6 strains of slideable bacteria producing anti-thrombotic active substances from water samples, soil samples and dead wood samples, and focused on strains B4 and B8. Using extraction method And (NH4) 2SO4 precipitation method were used to isolate the extracellular active proteins of strains B4 and B8. On the one hand, we detected the extracellular proteins of strains B4 and B8 by fibrin plate method and heated fibrin plate method, which could dissolve the thrombus by direct degradation of fibrin, detected by fibrinogen plate method and thrombin plate method The extracellular proteins of strains B4 and B8 could prevent thrombus formation by degrading fibrinogen. On the other hand, we used morphological characteristics, physicochemical properties, 16S rRNA sequence homology and phylogenetic tree to identify strains B4 and B8 Belong to Bacillus thuringiensis. In summary, the extracellular proteins of Bacillus thuringiensis B4 and B8 have both thrombolytic activity and anticoagulant activity, that is, they have antithrombotic effect.