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Liquid medium for producing plasmin by utilizing cordyceps sobolifera and plasmin production method thereof

A technology of liquid culture medium and fibrinolytic enzyme, which is applied in the field of microbiology, can solve the problems that Paecilomyces cicadae has not been reported, and achieve the effects of easy preparation, simple production process and simple formula

Inactive Publication Date: 2017-02-08
李伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although plasminase has been reported in other Cordyceps fungi such as Cordyceps sinensis and cordyceps militaris, it has not been reported in Paecilomyces cicadae

Method used

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  • Liquid medium for producing plasmin by utilizing cordyceps sobolifera and plasmin production method thereof
  • Liquid medium for producing plasmin by utilizing cordyceps sobolifera and plasmin production method thereof
  • Liquid medium for producing plasmin by utilizing cordyceps sobolifera and plasmin production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A kind of screening and cultivation method of the liquid culture medium that utilizes cicada flower to produce fibrinolytic enzyme:

[0033] 1 Medium used

[0034] (1) Basic medium: glucose 30g, potassium dihydrogen phosphate 1g, yeast extract 3g, magnesium sulfate heptahydrate 0.2g. 1000g of water. 1000 g of water is equivalent to 1000 mL of water.

[0035] (2) Liquid induction medium: Add 10g of the following ingredients to the basic medium: peptone, acid hydrolyzed casein, tryptone, starch, casein, malt extract ( malt extract), milk powder, soytone (Soytone), and made into different culture media for inducing plasmin. Routine high-temperature sterilization (1.1 atmospheres, 121°C for 20 minutes).

[0036] (3) Potato dextrose agar medium (potato dextrose agar, PDA): Cut 200g of potatoes into small pieces, add water and boil for 15min, then filter through 2 layers of gauze, discard the potato pieces, add 20g of glucose and 20g of agar to the filtrate Add distilled...

Embodiment 2

[0059] A kind of optimization of the liquid culture medium that utilizes cicada flower to produce plasmin:

[0060] 1 Medium used

[0061] (4) Basic medium: glucose 30g, potassium dihydrogen phosphate 1g, yeast extract 3g, magnesium sulfate heptahydrate 0.2g. 1000g of water. 1000 g of water is equivalent to 1000 mL of water.

[0062] (5) Liquid induction medium: Add different tryptones in the basic medium according to the following gradient: 5g, 7.5g, 10g, 12.5g, 15g, 17.5g, 20g, and make different induction fibrinolysis Enzyme medium. Routine high-temperature sterilization (1.1 atmospheres, 121°C for 20 minutes).

[0063] (6) Potato dextrose agar medium (potato dextrose agar, PDA): Cut 200g of potatoes into small pieces, add water and boil for 15min, then filter through 2 layers of gauze, discard the potato pieces, add 20g of glucose and 20g of agar to the filtrate Add distilled water to make up to 1000 mL; then perform conventional high-temperature sterilization (1.1 at...

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Abstract

The invention discloses a liquid medium for producing plasmin by utilizing cordyceps sobolifera and a plasmin production method thereof and belongs to the technical field of microbes. The liquid medium comprises the following active ingredients in parts by weight: 8-15 parts of tryptone, 30 parts of glucose, 1 part of monopotassium phosphate, 3 parts of yeast extract, 0.2 part of magnesium sulfate heptahydrate and 1000 parts of water. The method for producing plasmin by utilizing the liquid medium for producing plasmin by utilizing cordyceps sobolifera comprises the following steps of: (1) inoculating a paecilomyces cicadae strain (B5) onto a PDA panel with the diameter of 9cm, and culturing at the temperature of 25 DEG C for 5 days; (2) filling 200mL of the prepared liquid medium into a 500mL triangular flask, sterilizing at the temperature of 121 DEG C for 20 minutes, cooling to room temperature, teasing a piece of 3*3mm<2> hypha by using a teasing needle, inoculating to the prepared liquid induced medium panel, and culturing on a shaker at the temperature of 25 DEG C for 6-15 days; and (3) performing high-speed centrifugal separation on the culture solution at a speed of 10000 revolutions per minute for 5 minutes, and taking the supernatant as plasmin crude extract for later use.

Description

technical field [0001] The invention relates to a liquid culture medium for producing fibrinolytic enzymes by using cicada flowers and a method for producing fibrinolytic enzymes, which belong to the technical field of microbes, and in particular to a liquid medium and a culture method for inducing Paecilomyces cicadae to produce fibrinolytic enzymes. Background technique [0002] Thrombotic disease is a common disease with a relatively high fatality rate, mainly including cerebral thrombosis, pulmonary embolism, coronary artery blood flow, venous thromboembolism, atherosclerotic thrombosis, peripheral arterial thrombosis, etc., which seriously threaten human health. The treatment of thrombotic diseases mainly includes surgical thrombectomy and drug thrombus removal, and now the main method is drug thrombolysis. According to statistics, there are more than 15 million patients with various thrombotic diseases in the world, and this disease is developing at a younger age, requ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/68C12R1/79
CPCC12N9/6435C12Y304/21007
Inventor 李伟
Owner 李伟
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