纤溶酶产生菌的分离筛选与鉴定

The isolation,screening and identification of strains producing fibrinolytic enzyme

目的分离筛选具有较高纤溶活性的产酶菌株,对产酶菌株进行初步鉴定,并进一步对产酶菌株的体外溶栓机制进行初步研究。方法用生理盐水浸提实验菌固体发酵的大豆制备纤溶酶粗酶,通过纤维蛋白平板法筛选具有较高纤溶活性的产酶菌株,并进一步对菌株的菌落形态、菌体特征和生理生化特征进行鉴定。同时采用加热纤维蛋白平板法对菌株产生的纤溶酶在体外水解纤维蛋白的方式进行研究。结果从采集的多个样品中分离筛选到两株具有较高纤溶活性的产酶菌株C1A-01和C1A-03,初步确定这两个菌株均属于枯草芽孢杆菌(Bacillus subtilis)。体外溶栓实验表明,C1A-01和C1A-03是通过直接溶解纤维蛋白的方式发挥溶栓作用,而不是通过激活纤溶酶原间接起作用。结论获得了两株产较高纤溶酶活性的枯草芽孢杆菌菌株C1A-01和C1A-03,而且溶纤方式特殊,有望开发成新一代的抗栓溶栓药物。

Objective To isolate,screen and identify the strains that can produce strong fibrinolytic enzymes.Methods Crude enzyme was extracted from fermented soybean by experimental bacteria with saline. The strains with strong fibrinolytic activities were screened by the fibrin plate method and identified by their morphological, physiological and biological characters. Meanwhile, pyrogenation fibrin plate method （heating up） was used to study the fibrinolytic enzymes hydrolyzing fibrin in vitro.Results Two strains producing strong fibrinolytic enzymes were isolated and screened from many samples. They were named as C1A-01 and C1A-03 and identified as Bacillus subtitis. In vitro, C1A-01 and C1A-03 can take effect by hydrolyzing fibrin directly rather than by activating plasminogen indirectly in our study. Conclusion We acquired two Bacillus subtitis strains C1A-01 and C1A-03 producing strong fibrinolytic enzyme, which posses thrombolytic activities in a special way in vitro. There seems to be prospects for researches to develop C1A-01 as an ideal medicine candidate for prevention and treatment of thrombns.