产高活性抗凝溶栓双活性蛋白的短小芽孢杆菌的筛选及鉴定

Screening and identification of Bacillus pumilus producing double active protein of anticoagulation and thrombolysis

【目的】筛选可产生抗血栓活性物质的细菌。【方法】利用VY/4平板、酪蛋白平板从水样、土样、兔粪、羊粪、朽木等20多个样品中筛选目的菌株;利用纤维蛋白平板和纤维蛋白试管检测抗血栓活性;利用形态学特征、理化性质、16S rRNA序列同源性鉴定目的菌株。【结果】得到5株可产生抗血栓活性物质的细菌,重点研究了菌株LDS33,发现其分泌的胞外蛋白在纤维蛋白平板上和纤维蛋白试管中均显示出强烈的溶栓活性,通过试管法发现此蛋白质同时具有较强的抗凝活性。结合形态学、理化性质、16S rDNA序列及进化树分析,发现该菌株属于硬壁菌门芽孢杆菌目芽孢杆菌科芽孢杆菌属的短小芽孢杆菌,将其命名为Bacillus pumilus LDS.33。【结论】短小芽孢杆菌LDS33可产生高活性的抗凝溶栓双活性蛋白。

［Objective］The aim of this study was to screen bacteria that can produce antithrombotic．［Methods］ We screened the target bacteria on VY/4 plate and casein plate from more than 20 samples such as water，soil，rabbit manure，sheep manure and deadwood．We detected the antithrombotic activity by fibrin plate and fibrin tube．We identified the target bacteria by morphological characteristics，physical and chemical properties and 16S DNA sequence homology.［Results］We obtained 5 strains that can produce antithrombotic．We found that the extracellular protein of strain LDS33 shows both stronger fibrinolytic activity and stronger anticoagulation activity．According to the morphology，physiochemical properties，16S DNA sequencing and phylogenetic tree，strain LDS33 is identified as Bacillus pumilus．［Conclusion］Bacillus pumilus LDS33 can produce highly active anticoagulation and thrombolysis double active protein.