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Culture method for increasing content of glycyrrhizic acid in liquorice suspension culture cells

A technology for suspension culture cells and culture methods, which is applied in the field of cultivation to increase the content of glycyrrhizic acid in suspension culture cells of Glycyrrhizae, can solve the problems of low yield of Glycyrrhiza cell cultures, cannot meet the needs of expanding cell culture to produce Glycyrrhizic acid, etc., and achieve gene Effect of expression increase, content increase, content enhancement

Active Publication Date: 2020-09-29
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the low yield of licorice cell culture in the prior art, which cannot meet the needs of expanded cell culture to produce glycyrrhizic acid, the present invention uses glycyrrhizic acid content and cell biomass as screening conditions, and specifically induces the increase of glycyrrhizic acid biosynthesis and accumulation. Research, screened out a culture method to increase the content of glycyrrhizic acid in licorice suspension culture cells

Method used

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Experimental program
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Effect test

Embodiment 1

[0017] A culture method for improving glycyrrhizic acid content in licorice suspension culture cells adopts the following steps:

[0018] (1) Obtain licorice aseptic seedlings and select licorice seeds with full grains, corrode them with concentrated sulfuric acid for 30 minutes, rinse them with clear water, then use 75% ethanol for surface disinfection for 30 seconds, wash them twice with sterile water, and then Soak in 1% sodium hypochlorite solution for 10 minutes, shake fully during the soaking period, and then wash with sterile water for 3 times, each time not less than 1 minute; inoculate the washed seeds with 30g. -1 In the MS basic medium of agar, the pH value is 6.0, and the temperature is 23°C, the light intensity is 2000Lux, and the light time is 16h / d, and it is cultivated for 25 days. The seeds germinate and grow, the hypocotyl grows to 2cm, and the cotyledons are unfolded to obtain licorice. sterile vaccine;

[0019] (2) Inducing the callus to select the licoric...

Embodiment 2

[0024]A culture method for increasing glycyrrhizic acid content in licorice suspension culture cells, comprising the steps of:

[0025] (1) Obtain licorice aseptic seedlings and select licorice seeds with full grains, corrode them with concentrated sulfuric acid for 60 minutes, rinse them with clear water, then use 75% ethanol for surface disinfection for 30 seconds, wash them twice with sterile water, and then Soak in 3% sodium hypochlorite solution for 20 minutes, shake fully during soaking, and then wash 5 times with sterile water, each time not less than 1 minute; inoculate the washed seeds into 30g.L-1 sucrose, 6g.L -1 In the MS basic medium of agar, the pH value is 6.0, and the temperature is 27°C, the light intensity is 4000Lux, and the light time is 16h / d, and the light conditions are cultivated for 30 days. The seeds germinate and grow, the hypocotyl grows to 3cm, and the cotyledons are unfolded to obtain licorice. sterile vaccine;

[0026] (2) Inducing the callus to...

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Abstract

The invention discloses a culture method for increasing content of glycyrrhizic acid in liquorice suspension culture cells. The method comprises culture steps as follows: acquiring aseptic liquorice seedlings, inducing callus, establishing a liquorice cell suspension culture system and inducing accumulation of glycyrrhizic acid in the suspension culture system cells. With adoption of the method, saccharose is supplementarily added on the eighth-tenth days of inducing an accumulation culture process of glycyrrhizic acid in the suspension culture system cells, methyl jasmonate is added at the final stage of culture, accumulation of glycyrrhizic acid is induced to increase the content of glycyrrhizic acid, on the basis of guaranteeing stable biomass of the liquorice suspension culture cells,yield of glycyrrhizic acid is increased by about 10 times as compared with a conventional method, besides, a large number of liquorice suspension culture cells with higher content of glycyrrhizic acidcan be obtained continuously, efficiently, stably and rapidly, the culture is not limited by seasons or natural ecological conditions, the liquorice cells can be cultured in a large scale for glycyrrhizic acid production, and the culture method has broad application prospects and remarkable social and economic benefits.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a culture method for increasing the content of glycyrrhizic acid in licorice suspension culture cells. Background technique [0002] Glycyrrhizic acid is the most important active ingredient in licorice, which has various pharmacological activities such as anti-inflammatory, anti-ulcer, anti-virus and immune regulation. It is also reported that glycyrrhizic acid has the effect of anti-SARS virus, and can enhance the immunity of SARS-infected patients. The update of separation and purification technology has gradually pushed the application of glycyrrhizic acid to modernization. The latest generation of licorice preparation diammonium glycyrrhizinate enteric-coated capsules is a drug with two national invention patents and unique preparation advantages. It is a specific treatment for various liver inflammations. drug. At the same time, because the sweetness of glycyrrhizic acid is 150 ti...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/02C12P33/20A01G22/40A01H4/00
CPCC12N5/04C12N5/0025C12P33/20A01G22/40A01H4/001
Inventor 陈彩霞李先恩王文全
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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