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A kind of bacillus, the method of using bacillus to produce plasmin and the application of plasmin in thrombolytic drugs

A technology of Bacillus sp. and Bacillus sp. is applied in the application of plasmin in thrombolytic drugs and the field of preparation of plasmin to achieve the effects of small molecular weight, good thrombolytic effect, and low risk of human immune response

Inactive Publication Date: 2019-04-19
辛雄 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microorganisms have always been an important source of plasmin, especially Bacillus bacteria, and dozens of different types of plasmin have been isolated, such as NK isolated from Bacillus Subtilis, DC4 isolated from Bacillus amyloliquefaciens, etc. Others such as Bacillus subtilisin Carlsberg , Bacillus cereus and other bacteria have been reported to isolate plasmin (in Peng, Yong, Xiaojuan Yang, and Yizheng Zhang. "Microbial fibrinolytic enzymes: an overview of source, production, properties, and thrombolytic activity in vivo." Applied microbiology and biotechnology 69.2 ( 2005): 126-132 reported), but the use of Bacillus tequilensis to separate and prepare plasmin has never been reported

Method used

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  • A kind of bacillus, the method of using bacillus to produce plasmin and the application of plasmin in thrombolytic drugs
  • A kind of bacillus, the method of using bacillus to produce plasmin and the application of plasmin in thrombolytic drugs
  • A kind of bacillus, the method of using bacillus to produce plasmin and the application of plasmin in thrombolytic drugs

Examples

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Effect test

Embodiment 1

[0023] Weigh 20 grams of soybeans and 2 g of MgCl 2 , add 10 ml of double distilled water, place in a 200 ml beaker, sterilize at 121°C for 20 minutes, and inoculate a ring of Bacillus tequilensis bacteria after cooling. Fermentation at 32°C for 72 h.

[0024] After the fermentation is complete, add 200 ml of sterile water to the culture medium, stir for 0.5 h, filter through 4 layers of gauze to remove bacteria and insoluble impurities, and centrifuge at 11500 rpm for 20 min to remove the remaining impurities. Slowly add 112 g of solid ammonium sulfate, stir evenly, and let stand at 4°C for 1 h until the enzyme is completely precipitated.

[0025] Precipitate by adding 45 ml Na 2 CO 3 / NaHCO 3 (30 mM pH 9.2) solution was completely dissolved, using a dialysis bag with a molecular weight of 5 kDa or 8 kDa in Na 2 CO 3 / NaHCO 3 (30 mM pH 9.2) solution for dialysis, and the dialysate was changed every 8 h for a total of three times to remove residual ammonium sulfate mole...

Embodiment 2

[0029] Weigh the weight of peptone: yeast powder: sodium chloride: magnesium chloride (2:1:2:1) and dissolve in 100 times the volume (w / v) of distilled water, and divide into 200 ml, 1 / 3 of the liquid volume In a Erlenmeyer flask, sterilize at 121°C for 20 min. After cooling, each bottle was inoculated with a ring of Bacillus tequilensis strain. Fermentation at 35°C for 72 hours.

[0030] After the fermentation is completed, filter the fermentation 4-layer gauze to remove bacteria and insoluble impurities, and the remaining impurities can be removed by centrifugation at 11500 rpm for 20 min. After cooling, slowly add 560 g of anhydrous ammonium sulfate powder per liter of fermentation broth, stir evenly, and stand at 4°C for 4 h until the enzyme is completely precipitated.

[0031] Precipitate with as little Na as possible 2 CO 3 / NaHCO 3 (30 mM pH 9.2) solution was completely dissolved, and the precipitate produced by 1 L of fermentation broth was dissolved with 15 ml of ...

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Abstract

The invention aims at providing bacillus, a preparation method of plasmin and application of the plasmin. The bacillus is named as bacillus tequilensis, which is preserved with preservation number of CGMCC No. 11462; the method for producing the plasmin comprises steps of conducting fermentation culture on the bacillus tequilensis (with preservation number of CGMCC No. 11462), and implementing separation and purification on a fermentation product so as to obtain the bacillus plasmin; and in thrombolytic drugs, the plasmin, prepared by the method, is used for activating plasminogen in blood so as to degrade fibrin. The invention is applicable to the fields of production and processing of food raw materials.

Description

technical field [0001] The invention belongs to the technical fields of fermentation engineering and biopharmaceuticals, and in particular relates to a bacillus subtilis, a method for preparing plasmin by using the bacterium and the application of plasmin in thrombolytic drugs. Background technique [0002] Thrombosis is a kind of disease that seriously endangers human health and life. It is estimated that there are about 15 million patients with thrombosis in the world, of which 12 million die from cardiovascular and cerebral thrombosis every year. According to the World Health Organization (WHO) According to statistics, the death caused by thrombotic diseases such as myocardial infarction and cerebral hemorrhage has accounted for 51% of the total death toll in the world. Disease has surpassed cancer and diabetes to become the number one health killer. Drugs for the treatment of thrombosis can be divided into according to their different mechanisms of action: one, plasmino...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/68A61K38/48A61P7/02C12R1/07
Inventor 辛雄雷波蔡宗苇
Owner 辛雄
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