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Method for preparing epigallocatechin-3-gallate and epigallocatechin gallate by liquid-submerged fermentation

A technology of epicatechin gallate and epigallocatechin, which is applied in the field of biotechnology or bioengineering, and can solve problems such as uneconomical, cumbersome purification process of the mixture, and high price

Inactive Publication Date: 2013-06-12
徐向群
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is mainly obtained from tea leaves. However, there are several problems in the extraction and separation of tea leaves: organic solvent extraction is required, the obtained mixture needs cumbersome purification process, resulting in high cost and environmental impact, and low yield.
The preparation method has the problem of residual harmful organic solvents (chloroform, dichloromethane); the cost of obtaining high-purity epigallocatechin gallate (EGCG) and the purification and preparation of epicatechin gallate (ECG) is high and expensive ; The darker color of the extract affects the application in beverages and high-end skin care products, etc.
However, plant tissue cell culture faces the problems of uneconomical and difficult large-scale cultivation. Therefore, after 10 years of efforts by the scientific community, there is no commercial large-scale system for producing tea catechins from plant tissue cells.

Method used

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  • Method for preparing epigallocatechin-3-gallate and epigallocatechin gallate by liquid-submerged fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The mycelia of Inonotus obliquus from the Dutch CBS strain collection were inoculated in the slant medium. The culture temperature was 28° C., and cultured for 240 hours. The fresh slant strains were inserted into a 500mL shake flask filled with 200mL liquid seed medium, cultured at 28°C for 96 hours, and the shaker rotated at 150 rpm. The cultured liquid seeds are inserted into a fermenter equipped with a rice straw fermentation medium according to 10% (v / v). Incubate at 28°C for about 288 hours. The air flow rate is 1.0vvm, the internal pressure is 0.2 kg / cubic centimeter, and the stirring speed is 140 rpm.

[0022] In Example 1, the slant strains were cultured, and the composition of the culture medium was (g / 100mL): 25 malt extracts, 4 peptones, 15 agar, and the pH value was 5.4-5.6.

[0023] Shake flask seed culture in this example 1, medium composition (g / 100mL): glucose 3, peptone 0.4, yeast extract 0.1, KH2PO4 0.1, MgSO4 0.15, CaCl2 0.01.

[0024] Submerged ...

Embodiment 2

[0029] The mycelia of Inonotus obliquus from the Dutch CBS strain collection were inoculated in the slant medium. The culture temperature was 28° C., and cultured for 240 hours. The fresh slant strains were inserted into a 500mL shake flask containing 200mL liquid seed medium, and cultured at 28°C for 96 hours with a shaker speed of 140 rpm. The cultivated liquid seeds are inserted into the fermenter equipped with the sawdust fermentation medium according to 10% (v / v). Incubate at 28°C for about 302 hours. The air flow rate is 0.5vvm, the internal pressure is 0.1 kg / cubic centimeter, and the stirring speed is 140 rpm.

[0030] In Example 2, the slant strains were cultured, and the composition of the culture medium was (g / 100mL): 25 malt extracts, 4 peptones, 15 agar, and the pH value was 5.4-5.6.

[0031] Shake flask seed culture in Example 2, medium composition (g / 100mL): glucose 3, peptone 0.4, yeast extract 0.1, KH2PO4 0.1, MgSO4 0.15, CaCl2 0.01. The shaker rotates at ...

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Abstract

The invention relates to a method for increasing yield of epigallocatechin-3-gallate (EGCG) and epigallocatechin gallate (ECG) by liquid-submerged fermentation of white-rot fungi by adding cellulose, hemi-cellulose and lignin, fatty acids, organic solvents and surfactants. Fermentation in a liquid fermentation tank is carried out with preparation of catechins by fermentation white-rot fungi such as inonotus obliquus as an object and Holland CBS culture collection inonotus obliquus as initial strains. Fermentation conditions are as follows: a temperature is 27-28 DEG C; an air flow is 0.5-1.5 vvm; a stirring speed is 80-160 rounds per minute; an internal pressure is 0.15-0.35 kilograms per cubic centimeter; and a fermentation cycle is 10-14 days. The method can increase the yield of epigallocatechin-3-gallate (EGCG) and epigallocatechin gallate (ECG) by liquid-submerged fermentation of the inonotus obliquus.

Description

technical field [0001] The invention relates to a process for preparing epigallocatechin gallate (EGCG) and epicatechin gallate (ECG) by submerged fermentation of white-rot fungus liquid, belonging to the field of biotechnology or bioengineering. technical background [0002] Epigallocatechin gallate (EGCG) and epicatechin gallate (ECG) are tea catechins with the strongest antioxidant activity and are widely used in medicine, food and cosmetic industries. At present, it is mainly obtained from tea leaves. However, there are several problems in the extraction and separation from tea leaves: organic solvent extraction is required, and the obtained mixture needs cumbersome purification process, resulting in high cost and environmental impact, and low yield. The preparation method has the problem of residual harmful organic solvents (chloroform, dichloromethane); the cost of obtaining high-purity epigallocatechin gallate (EGCG) and the purification and preparation of epicatechin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/06C12R1/645
Inventor 徐向群
Owner 徐向群
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