Method for acquiring secondary metabolites including panax japonicus saponins and the like with large-scale culture and bioconversion technology for panax japonicus cells

A bamboo ginseng saponin and biotransformation technology, applied in the field of compounds, can solve problems such as difficult reproduction, limited growth, and difficult artificial cultivation

Active Publication Date: 2016-07-13
郭志刚 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although artificial cultivation has been carried out in Jiangxi, Hunan, Hubei and other places, it has been difficult to realize large-...

Method used

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  • Method for acquiring secondary metabolites including panax japonicus saponins and the like with large-scale culture and bioconversion technology for panax japonicus cells
  • Method for acquiring secondary metabolites including panax japonicus saponins and the like with large-scale culture and bioconversion technology for panax japonicus cells
  • Method for acquiring secondary metabolites including panax japonicus saponins and the like with large-scale culture and bioconversion technology for panax japonicus cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0038] (1) In the RW medium, add α-naphthyl acetic acid (NAA), 2,4-D or indole acetic acid (IAA) 2mg / liter and 6-bean aminopurine (6-BA) or kinetin (KT) 4mg / L, sucrose or glucose 20g / L, then adjust the pH value to 5.6 with acid or alkali, then add 2g / L gellan gum as a coagulant, and sterilize at 115-120°C and 0.1MPa pressure for 18 minutes, Cooled to make slant induction medium;

[0039] (2) Sterilize the young stems or buds of Panax ginseng as explants on the surface of 70% ethanol for 50 seconds, then sterilize them in 5% sodium hypochlorite or calcium hypochlorite solution for 20 minutes, rinse with sterile water three times , cut off the surface-sterilized shoot tips under sterile conditions, or cut the young stems into 0.5 cm or take the shoot tips and inoculate them in the above-mentioned slant induction medium. Then transfer to 25°C, 2000 Lux, 12 hours of light conditions for induction culture, and callus will form after 2 months;

[0040] (3) In the RW medium, add 1 ...

example 2

[0051] (1) In the RW medium, add α-naphthaleneacetic acid (NAA) 5mg / liter and 6-bean aminopurine (6-BA) 5mg / liter, sucrose or glucose 20g / liter, and then adjust the pH value with acid or alkali Adjust it to 5.5-7.0, then add 5g / L agar as a coagulant, sterilize at 115-120°C and 0.1MPa pressure for 15 minutes, and cool to make a slant induction medium;

[0052] (2) Sterilize the young stems or buds of Panax ginseng as explants on the surface of 75% ethanol for 30 seconds, then sterilize them in 6% sodium hypochlorite or calcium hypochlorite solution for 15 minutes, and rinse them twice with sterile water Finally, cut off the surface-sterilized shoot tip under aseptic conditions, or cut the young stem into 0.8 cm or get its shoot tip and inoculate it in the above-mentioned slant induction medium. Then transfer to 18°C, 2000 Lux, 12 hours of light conditions for induction culture, and callus will form after 2 months;

[0053] (3) Add 0.1mg / L indole acetic acid (IAA), 5mg / L and 6-...

example 3

[0064] (1) In the RW medium, add 2,4-D 0.5mg / L and kinetin (KT) 1mg / L, sucrose 30g / L, then adjust the pH value to 5.5 with acid or alkali, and then add 1.8g / L gellan gum is used as a coagulant, and after being sterilized at 115-120°C and 0.1MPa pressure for 18 minutes, it is cooled to make a slant induction medium;

[0065] (2) Sterilize the young stems or buds of Panax ginseng as explants on the surface of 70% ethanol for 40 seconds, then put them into 6% sodium hypochlorite solution for sterilization for 15 minutes, wash them three times with sterile water, and then dry them under aseptic conditions. Next, cut off the shoot tips after surface sterilization, or cut the young stems into 0.8 cm or get the shoot tips and inoculate them in the above-mentioned slant induction medium. Then transfer to 23°C, 2500 Lux, 14 hours of light conditions for induction culture, callus formed after 1 month;

[0066] (3) In the RW medium, add 2mg / L2, 4-D, 2mg / L and 6-bean aminopurine (6-BA),...

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Abstract

The invention relates to the field of plant cell culture and provides a method for acquiring secondary metabolites including panax japonicus saponins and the like with large-scale culture and a bioconversion technology for panax japonicus cells. The method comprises steps as follows: (1), auxins, cytokinins and a coagulator are added to an RW culture medium; (2), explant surface of panax japonicas is sterilized and callus is formed through induction; (3), the callus of the panax japonicas is subjected to subculture, and loose cell clusters are obtained; (4), a culture solution is prepared; (5), the loose cell clusters are transferred into the culture solution for culturing; (6), high-yield cell strains are cultured and screened; (7), the high-yield cell strains are subjected to multiplication culture; (8), a synthetic medium is prepared; (9), cells are continuously cultured in the synthetic medium, and bioconversion is completed; (10), purification is performed after extraction with a solvent. With the adoption of the method, the secondary metabolites of the panax japonicas can be produced, the production cost is low, the culture period is short, the method is not limited by natural environment, and annual production can be realized.

Description

technical field [0001] The invention relates to a method for obtaining secondary metabolites of Panax ginseng, in particular to a method for obtaining compounds such as Panax ginseng saponins by using large-scale cultivation and biotransformation technology of Panax ginseng cells. Background technique [0002] Bamboo ginseng (Panax japonicus C.A. Mey), also known as bamboo ginseng or white notoginseng, is a plant of the genus Panax japonicus. Commonly known as "herb king". Mainly distributed in my country's Jiangxi, Hubei, Guangxi, Sichuan, Guizhou, Yunnan, Tibet and other provinces and regions under alpine bushes, wetlands or beside rocky ditches. It has many different names. The "Dictionary of Chinese Medicine" says: Different books have different titles, such as: soil ginseng, soil essence, blood ginseng ("Hua Jing"), sweet seven, bamboo knot seven ("Herbs") "Convenience Recipe"), Bamboo Ginseng ("Scientific Folk Herbs"), Bamboo Whip Sanqi, Arhat Notoginseng ("Chinese M...

Claims

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Application Information

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IPC IPC(8): C12P33/20
CPCC12P33/20
Inventor 郭志刚弭博旌
Owner 郭志刚
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