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Method for producing cordycepin with self-assembly silicon plastid electrostatic spinning fibrous membrane immobilized cells

A technology for cell production of cordycepin and electrospinning, applied in microorganism-based methods, biochemical equipment and methods, immobilized on or in inorganic carriers, etc., can solve complex synthesis processes, difficult product purification, and high production costs. problem, to achieve the effect of high reaction efficiency, low diffusion resistance and high porosity

Active Publication Date: 2018-02-13
芽米科技(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high production cost of chemically synthesized cordycepin, complex synthesis process, low yield and difficult product purification, cordycepin is mainly prepared by biosynthesis

Method used

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  • Method for producing cordycepin with self-assembly silicon plastid electrostatic spinning fibrous membrane immobilized cells
  • Method for producing cordycepin with self-assembly silicon plastid electrostatic spinning fibrous membrane immobilized cells
  • Method for producing cordycepin with self-assembly silicon plastid electrostatic spinning fibrous membrane immobilized cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The preparation of Cordyceps militaris protoplasts, the steps are as follows:

[0038] (1) Take 3 g of Cordyceps militaris cells, wash them thoroughly with sterile water, and centrifuge to get the precipitate.

[0039](2) Preparation of enzymolysis solution: take 0.1g of lysozyme, 0.1g of helicase, 0.1g of cellulase, and 0.1g of collapsing enzyme, and add them to the solution containing 0.3mol / L KCl and 0.4mol / L MgSO 4 10mL aqueous solution, fully dissolved, and then sterilized by filtration to obtain an enzymolysis solution.

[0040] (3) The precipitate was added to the enzymatic hydrolysis solution, vortexed to mix, enzymatic hydrolyzed in a water bath at 30°C for 8 hours, centrifuged, and the precipitate was the Cordyceps militaris biomass.

Embodiment 2

[0042] The preparation of Cordyceps militaris protoplasts, the steps are as follows:

[0043] (1) Take 4 g of Cordyceps militaris cells, wash them thoroughly with sterile water, and centrifuge to get the precipitate.

[0044] (2) Preparation of enzymolysis solution: take 0.1g of lytic enzyme, 0.1g of helicase, 0.1g of cellulase, and 0.1g of collapsing enzyme, and add them to the solution containing 0.5mol / L KCl and 0.2mol / L MgSO 4 5mL aqueous solution, fully dissolved, and then sterilized by filtration to obtain an enzymatic hydrolysis solution.

[0045] (3) The precipitate was added to the enzymatic hydrolysis solution, vortexed to mix, enzymolyzed in a water bath at 25°C for 3 hours, centrifuged, and the precipitate was the Cordyceps militaris biomass.

Embodiment 3

[0047] (1) Weigh 150mg of organic-inorganic composite lipid and dissolve it in 10mL of chloroform, remove the chloroform by rotary evaporation, and form a transparent lipid-like film at the bottom of the round-bottomed flask; Suspend 180g / L mannitol aqueous solution, add to the obtained lipid film, stir well and rapidly heat to 48°C, heat shock for 20s and ultrasonic in water bath; rapidly cool to 4°C, stand for 12h, and extruded through a body extruder (polycarbonate filter membrane with a pore size of 1.0 μm) to obtain a siliceous body suspension of Cordyceps militaris biomass with uniform particle size.

[0048] (2) Take chitosan (viscosity-average molecular weight 5.0×10 5 , deacetylation degree 85%), polyvinyl alcohol (weight average molecular weight is 200000) and polyethylene oxide (weight average molecular weight is 1×10 7 ) is completely dissolved in the acetic acid aqueous solution of 2% with the total mass concentration of 1.8%, adding the glycerol with the mass vo...

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Abstract

The invention discloses a method for producing cordycepin with self-assembly silicon plastid electrostatic spinning fibrous membrane immobilized cells. The method comprises the following steps that Chinese caterpillar fungus protoplasts are wrapped into a stable silicon plastid formed by organic-inorganic hybrid lipids by utilizing a electrostatic self-assembly method; chitosan, polyvinyl alcoholand polyethylene oxide are completely dissolved into an acetic acid solution, glycerol is added, then silicon plastid is added, a nanometer fibrous membrane is prepared with electrostatic spinning, the nanometer fibrous membrane is rolled, and the rolled nanometer fibrous membrane is charged in a chromatography column; a mixed solution of glucose, ribose, potassium dihydrogen phosphate, magnesiumsulfate, yeast powder and adenine is prepared, filtering and bacteria removal are carried out on the mixed solution, the mixed solution flows through the chromatography column at a certain flow rate,gas is introduced at the lower part of the chromatography column at the same time, and through detection, an effluent contains the cordycepin. The method with the immobilized cells has the characteristics of high efficiency, bacterium contamination resistance, high yield and high concentration of a product and is beneficial to separation and purification of a subsequent product.

Description

technical field [0001] The invention relates to a method for producing cordycepin by self-assembled siliceous body electrostatic spinning fiber membrane immobilized cells, belonging to the field of biochemical industry. Background technique [0002] Cordycepin is one of the main active ingredients in Cordyceps. Cordycepin has a variety of biological activities, such as anti-tumor, anti-proliferation, anti-metastasis, antibacterial, antiviral, immune regulation, and anti-inflammation. The preparation of cordycepin mainly includes chemical synthesis and biosynthesis. Due to the high production cost of chemically synthesized cordycepin, complex synthesis process, low yield and difficult product purification, cordycepin is mainly prepared by biosynthesis. There are two ways to prepare cordycepin by biosynthesis: one is to obtain Cordyceps fruiting body by solid fermentation, and then extract it; the other is to directly extract from the fermentation broth through liquid fermen...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10C12N11/08C12N11/04C12N1/14C12P19/40D04H1/4382D04H1/728C12R1/645
CPCC12N1/14C12N11/04C12N11/08C12N11/10C12N11/14C12P19/40D04H1/4382D04H1/728
Inventor 汤佳鹏葛彦姜正林
Owner 芽米科技(广州)有限公司
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