Preparation method for bacterial-originated defibrase

Abstract

The present invention discloses a preparation method for bacterial-originated defibrase. According to the method, strains of enterococcus faecalis EF608 are adopted as the extract; the strains of the enterococcus faecalis EF608 are subjected to fermentation culture; the resulting fermentation broth is subjected to centrifugation to remove the precipitate; solid ammonium sulfate is added to the resulting supernatant until the saturation of the ammonium sulfate is 60-65%; the resulting salting-out component is subjected to dialysis and concentration to obtain a crude extracting solution; the resulting crude extracting solution is sequentially treated by a phenyl-agarose gel FF hydrophobic chromatography column, a DEAE-SephadexA-25 ion exchange chromatography column and a Sepharose-heparin affinity chromatography column to obtain the component having a high abundance fibrinolytic activity peak; the resulting component is subjected to treatments of dialysis and freeze drying to obtain the pure product, wherein the pure product is the bacterial-originated defibrase. With the preparation method provided by the present invention, the fibrinogenase having a molecular weight of 37.1 KD is firstly separated and purified from the fermentation broth of the strains of the enterococcus faecalis; the fibrinogenase is easy to be purified and prepared batchly; the fibrinogenase has strong fibrinolytic activity and no toxicity, and is an ideal raw material for preparing the fibrinogenase.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing defibrase by taking bacterial sources as extracts. Background technique [0002] Thrombus is a mass formed by the blood itself in the vascular system. Many lesions caused by thrombus occlusion of the pipeline are thromboembolic diseases, such as myocardial infarction, cerebral thrombosis, pulmonary thrombosis, and venous thrombosis. With the prolongation of human life span and the change of diet structure, the cases of thromboembolism have also increased significantly. According to the survey of the World Health Organization, there are no less than 15 million patients with thromboembolic diseases in the world, and thromboembolic diseases have become a current hazard. Human health is one of the diseases with the highest mortality rate. [0003] Currently, one of the most reliable and effective means of treating thromboembolic diseases is thrombolytic therapy, t...

AI Technical Summary

Problems solved by technology

However, some of these strains have low enzyme production, some have pathogenicity, and the defibrase produced by some strains has relatively large side effects after clinical application. These factors restrict the development of microbial plasminase to a certain extent. Therefore, it is particularly important to screen out strains that are non-pathogenic, easy to cultivate, and have high enzyme production, and to find out mature process conditions

Images