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Method for producing plasmin through microbial fermentation

A microbial fermentation and fibrinolytic enzyme technology, applied in the fields of microbiology, biochemistry, enzyme engineering, and fermentation engineering

Inactive Publication Date: 2012-11-14
DALIAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the industrialization, scale and application of microbial fermentation to produce plasmin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] (1) Culture medium preparation

[0013] ① Strain activation medium: glucose 10.0g, fibrin 20.0g, distilled water 1.0L, pH value 7.0-7.2, sterilized by high-pressure steam at 121℃ for 30min.

[0014] ② Strain-directed acclimation medium: glucose 8.0-12.0g, fibrin 15.0-25.0g, yeast powder 3.0-6.0g, NaCl 4.0-6.0g, agar powder 20.0g, distilled water 1.0L, natural pH, high pressure at 121℃ Steam sterilization for 30min.

[0015] ③Liquid seed medium: glucose 5.0~15.0g, soybean peptone 6.0~12.0g, K 2 HPO 4 1.0~6.0g, KH 2 PO 4 0.5~2.5g, CaCl 2 0.1~0.5g, MgSO 4 0.2~0.8g, natural pH value, 1.0L tap water, sterilize by high-pressure steam at 121℃ for 30min.

[0016] ④ Fermentation enzyme production medium: glucose 15.0-25.0g, soybean milk 0.01-0.03L, yeast powder 1.0-6.0g, KH 2 PO 4 0.5~2.0g, K 2 HPO 4 1.5~6.5g, MgSO 4 ·7H 2 O 0.2~0.8g, CaCl 2 0.01~0.05g, natural pH value, 1.0L tap water, sterilized by high-pressure steam at 121℃ for 30min.

[0017] (2) The m...

Embodiment 2

[0023] (1) Culture medium preparation

[0024] ① Strain activation medium: glucose 10.0g, fibrin 20.0g, distilled water 1.0L, pH value 7.0-7.2, sterilized by high-pressure steam at 121℃ for 30min.

[0025] ② Strain-directed acclimation medium: glucose 8.0-12.0g, fibrin 15.0-25.0g, yeast powder 3.0-6.0g, NaCl 4.0-6.0g, agar powder 20.0g, distilled water 1.0L, natural pH, high pressure at 121℃ Steam sterilization for 30min.

[0026] ③Liquid seed medium: glucose 5.0~15.0g, soybean peptone 6.0~12.0g, K 2 HPO 4 1.0~6.0g, KH 2 PO 4 0.5~2.5g, CaCl 2 0.1~0.5g, MgSO 4 0.2~0.8g, natural pH value, 1.0L tap water, sterilize by high-pressure steam at 121℃ for 30min.

[0027] ④ Fermentation enzyme production medium: glucose 15.0-25.0g, soybean milk 0.01-0.03L, yeast powder 1.0-6.0g, KH 2 PO 4 0.5~2.0g, K 2 HPO 4 1.5~6.5g, MgSO 4 ·7H 2 O 0.2~0.8g, CaCl 2 0.01~0.05g, natural pH value, 1.0L tap water, sterilized by high-pressure steam at 121℃ for 30min.

[0028] (2) The m...

Embodiment 3

[0034](1) Culture medium preparation

[0035] ① Strain activation medium: glucose 10.0g, fibrin 20.0g, distilled water 1.0L, pH value 7.0-7.2, sterilized by high-pressure steam at 121℃ for 30min.

[0036] ② Strain-directed acclimation medium: glucose 8.0-12.0g, fibrin 15.0-25.0g, yeast powder 3.0-6.0g, NaCl 4.0-6.0g, agar powder 20.0g, distilled water 1.0L, natural pH, high pressure at 121℃ Steam sterilization for 30min.

[0037] ③Liquid seed medium: glucose 5.0~15.0g, soybean peptone 6.0~12.0g, K 2 HPO 4 1.0~6.0g, KH 2 PO 4 0.5~2.5g, CaCl 2 0.1~0.5g, MgSO 4 0.2~0.8g, natural pH value, 1.0L tap water, sterilize by high-pressure steam at 121℃ for 30min.

[0038] ④ Fermentation enzyme production medium: glucose 15.0-25.0g, soybean milk 0.01-0.03L, yeast powder 1.0-6.0g, KH 2 PO 4 0.5~2.0g, K 2 HPO 4 1.5~6.5g, MgSO 4 ·7H 2 O 0.2~0.8g, CaCl 2 0.01~0.05g, natural pH value, 1.0L tap water, sterilized by high-pressure steam at 121℃ for 30min.

[0039] (2) The mi...

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Abstract

The invention relates to a method for producing plasmin through microbial fermentation. According to the method, microbes for generating plasmin are activated for 6 to 10 times and are subjected to product directional domestication for 4 to 6 times so that the microbes well grow under the environment condition being 24 to 30 DEG C; plasmin producing bacteria subjected to product directional domestication by a conventional method are subjected to step-by-step enlargement culture at 24 to 30 DEG C and are inoculated into a fermentation culture medium at the inoculation quantity being 3 to 9 percent of the fermentation liquid volume, the culture is carried out for 60 to 114 hours at 24 to 30 DEG C, and the plasmin production through microbial fermentation is completed; the fermentation liquid is centrifuged at 4000 to 8000 rpm, liquid is collected, and the collected liquid is crude enzyme liquid; and according to different requirements and use objects, the crude enzyme liquid can be further concentrated, separated and purified, and enzyme preparations with different activities, purities and preparation forms are prepared.

Description

technical field [0001] The invention relates to the fields of microbiology, enzyme engineering, fermentation engineering, biochemistry and the like, in particular to a method for producing plasmin by microbial fermentation. The plasmin produced by this invention can directly degrade fibrin, and is mainly used in medical treatment, health care, and other plasmin application industries. It can be developed as functional food and new drugs for the treatment of cardiovascular and cerebrovascular related diseases, and improve cardiovascular and cerebrovascular embolism. The treatment process, efficacy and safety of other diseases. Background technique [0002] Plasmin (EC 3.4.21.7) refers to a proteolytic enzyme that can specifically degrade fibrin gel, and is an important component of the fibrinolytic system. Plasmin mainly degrades fibrin by degrading fibrin and fibrinogen, hydrolyzing various coagulation factors (II.V.VII.VIII.X.XI), converting plasminogen into plasmin, and h...

Claims

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Application Information

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IPC IPC(8): C12N9/68
Inventor 张庆芳迟乃玉窦少华王晓辉于爽袁慎亮
Owner DALIAN UNIVERSITY
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