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Yellow-green halimasch fibrinolytic enzyme and production method thereof

A technology of Armillaria yellow-green and a production method, applied in the field of Armillaria yellow-green and its production and preparation, to achieve the effect of easy extraction and purification

Inactive Publication Date: 2008-01-30
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Armillaria chrysogenum is currently in a completely wild state, domestication and cultivation research is still in its infancy, and its further development and utilization still needs further research

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Medium, culture method and extraction:

[0042] The strains were stored on a slant in PDA medium. First, the strains were cultured at 23° C. for 72 hours, and then inoculated on the liquid fermentation medium.

[0043] The liquid fermentation medium contains 2% glucose, 2% yeast extract, 10% soybean juice (preparation method: soak 10% soybeans for 24 hours, simmer for 30 minutes, and extract the juice), KH 2 PO 4 1%, MgSO 4 0.45%, pH 4.

[0044] Inoculum size 1×10 8 spores / ml, the fermentation temperature is 28°C, the shaker speed is 140r / min, and the maximum enzyme activity obtained is 310IU / ml after culturing for 120 hours.

[0045] Rough purification of enzymes: The fermented liquid is subjected to high-speed refrigerated centrifugation at a speed of 12000r / min and a temperature of 4°C to remove the bacteria to obtain supernatants, which are respectively 45% and 80% ammonium sulfate solutions by mass Carry out segmental salting-out, and dissolve the precipitate...

Embodiment 2

[0048] The strains were stored on a slant in PDA medium. First, the strains were cultured at 23° C. for 72 hours, and then inoculated on the liquid fermentation medium.

[0049] The liquid fermentation medium contains 2% bran, 2% yeast extract, 10% soybean juice (preparation method: soak 10% soybeans for 24 hours, simmer for 30 minutes, filter to obtain juice), KH 2 PO 4 1%, MgSO 4 0.45%, 0.1% CaCl 2 , pH 4. Inoculum size 1×10 8 spores / ml, the fermentation temperature was 28°C, the shaker speed was 140r / min, and the maximum enzyme activity obtained after 120 hours of fermentation was 450 IU / ml.

[0050] According to the rough purification of enzyme in embodiment 1 and the fine purification method of enzyme, obtain the higher Armillaria chrysanthemum fibrinolytic enzyme of enzyme activity, through SDS-PAGE analysis, its apparent molecular size is 26KDa; The ratio of this enzyme The activity is 40000 IU / mg protein.

Embodiment 3

[0052] The strains were stored on a slant in PDA medium. First, the strains were cultured at 23° C. for 72 hours, and then inoculated on the liquid fermentation medium.

[0053] The liquid fermentation medium contains 5% bran, 1% yeast extract, 10% soybean juice (preparation method: soak 10% soybeans for 24 hours, simmer for 30 minutes, filter to obtain juice), KH 2 PO 4 1%, MgSO 4 0.5%, 0.1% CaCl 2 , pH 4. The inoculum size is 1×10 8 spores / ml, the fermentation temperature is 28°C, the shaker rotation speed is 140r / min, and the maximum enzyme activity obtained by fermentation for 120 hours is 517IU / ml.

[0054] According to the rough purification of enzyme in embodiment 1 and the fine purification method of enzyme, obtain the higher Armillaria chrysanthemum fibrinolytic enzyme of enzyme activity, through SDS-PAGE analysis, its apparent molecular size is 26KDa; The ratio of this enzyme The activity is 50000IU / mg protein.

[0055] Example 3

[0056] The strains were st...

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Abstract

The invention discloses an armillaria luteo-virens fibrinolytic enzyme and a production method thereof. Fermentation and cultivation are made to armillaria luteo-virens ZJUQH CGMCC No.1884 to get fermented liquid and the armillaria luteo-virens fibrinolytic enzyme is separated and purified from the fermented liquid. The invention first realizes that artificial cultivated armillaria luteo-virens ZJUQH CGMCC No.1884 can be used for producing the fibrinolytic enzyme. The produced armillaria luteo-virens fibrinolytic enzyme takes agricultural and sideline products such as bean pulp, soybean flour and wheat bran with low cost as the main raw materials, thereby being safe and nonpoisonous. Being produced with a liquid fermentation method, the armillaria luteo-virens fibrinolytic enzyme is a hyper strain with relatively higher yield among the reports of epiphytes. Extraction and purification can be carried out easily and the armillaria luteo-virens is harmless, frozen-dry health products that contain thalli and enzyme liquid can be directly developed, thus providing a new thought and an evidence for developing and using novel edible fungus used in foods and medicines.

Description

technical field [0001] The patent of the invention belongs to the field of bioengineering, and mainly relates to a strain of Armillaria chrysanthemum producing plasmin and its production and preparation method. Background technique [0002] Edible and medicinal fungi have a long history in my country as medicine. As early as the end of the Eastern Han Dynasty more than 2,000 years ago, the world's first medicinal monograph "Shen Nong's Materia Medica" recorded the medicinal effects of fungi such as Ganoderma lucidum, Huoquan, and pigs; Li Shizhen, a famous medical scientist in the Ming Dynasty, wrote in "Compendium of Materia Medica )" contains more than 20 kinds of fungi. The existing research results show that there are nearly 300 species of fungi known to have an inhibitory rate of 60% to 100% against sarcoma and Ehrlich’s carcinoma, belonging to more than 70 genera in 51 families. The antitumor activity of most fungi is due to their specific structure. polysaccharides ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/68
Inventor 陈启和何国庆朱建良阮晖章海锋
Owner ZHEJIANG UNIV
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