Method for producing Syringin, chlorogenic acid and 1, 5-dicaffeoylquinic acid by cell culture of herba saussureae involucratae

A technology of caffeoylquinic acid and syringin, which is applied in the directions of biochemical equipment and methods, chemical instruments and methods, plant cells, etc., can solve the problems of non-disclosure, etc., and achieves strong production capacity, stable and controllable quality, and ease of use. The effects of the resource crisis

Inactive Publication Date: 2013-09-18
INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the prior art does not disclose a method for simultaneously producing three active

Method used

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  • Method for producing Syringin, chlorogenic acid and 1, 5-dicaffeoylquinic acid by cell culture of herba saussureae involucratae
  • Method for producing Syringin, chlorogenic acid and 1, 5-dicaffeoylquinic acid by cell culture of herba saussureae involucratae
  • Method for producing Syringin, chlorogenic acid and 1, 5-dicaffeoylquinic acid by cell culture of herba saussureae involucratae

Examples

Experimental program
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Effect test

Embodiment 1

[0079] Example 1, Screening of Saussurea sativa cell line with high yield of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid

[0080] The tissues and organs derived from wild Saussurea tianshanensis plants, regenerated seedlings or aseptic seedlings after germination are used as explants, preferably aseptic seedling leaves or hypocotyls after seed germination, to induce the production of isolated tissue cells, and the induction medium is Additional 0.1~2.0mg·L -1 NAA, 0.1~2.0mg·L -1 6-BA and 0.1~2.0mg·L -1 2,4-D hormone combination, and subculture in the same composition of medium to screen high-producing cell lines. For the subculture, the loose and light yellow tissue cells with good growth status were selected as seeds, and the whole cultivation process was carried out in the dark, subcultured once every 10 days, combined with the analysis of secondary metabolite components to screen and cultivate for more than 1 year, and high-yield syringin, syringin, and Chlo...

Embodiment 2

[0084] Example 2, the establishment of a high-yield syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid Saussurea sauraceae cell suspension system

[0085] The Saussurea tianshanensis cell line with high yields of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid obtained through screening was transferred into liquid medium for suspension culture in shake flasks. The bed is rotated and shaken for culture, and the speed of the shaker is 80-140r·min -1 , the inoculum amount is 5-10 g dry weight of cells per liter of medium, and the suspension culture is carried out under dark conditions. Subculture once every 8d to 10d, after 3 to 4 generations, a stable and high-yield syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid suspension cell culture system of Saussurea chinensis was established, and the cells in the suspension system showed the Granular, good dispersion, rapid growth, the biomass per liter of medium can reach 25g~30g dry cell weight.

[0086] Prepar...

Embodiment 3

[0087] Embodiment 3, the cell suspension system of Snow Lotus Tianshan is used for mass production of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid

[0088] By adding phenylpropanoids to synthesize precursor L-phenylalanine at the initial stage of culture, and adding elicitor methyl jasmonate during the culture process, the production of syringin, chlorogenic acid and 1 in Snow Lotus cells can be improved, 5-dicaffeoylquinic acid capacity.

[0089] Preparation and culture method of production medium: the production of active ingredients adopts additional 0.1-2.0mg·L -1 NAA, 0.1~2.0mg·L -1 6-BA and 0.1~2.0mg·L -1 2, 4-D hormone combination, and add 3% to 8% sucrose, add 10 to 100mg·L on the 4th to 8th day of culture -1 Methyl jasmonate, harvested on the 12th to 14th day of cultivation, the biomass per liter of the medium can reach 21g to 26g, and the yields of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid can reach 500 to 500g respectively by HPLC analy...

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Abstract

The invention provides a method for simultaneously producing and preparing three active constituents, including Syringin, chlorogenic acid and 1, 5-dicaffeoylquinic acid by cell culture of herba saussureae involucratae on large scale. The method comprises application of flask shaking suspension culture and bioreactor culture, and separation and preparation process for the three constituents in the cultured cell. The invention provides a new method for production of the three medicinal active constituents, and the new method can provide enough resource for clinical medicine application; moreover, the method can protect the rare wild plants resource while realizing harmless, pollution-free and low-cost large scale production of the herba saussureae involucratae active constituents, so that the resources crisis is eased effectively.

Description

technical field [0001] The invention relates to a method for producing medicinal active ingredients syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid by culturing cells of Snow Lotus Tianshan, and preparing the three active ingredients respectively by chromatography. Background technique [0002] Plant cells are totipotent, a plant cell has the same genetic information as the parent plant, not only has the ability to develop into a complete plant, but also has the ability to synthesize compounds that the whole plant has, and the parent plant can be obtained through plant cell culture The main medicinal active ingredients, which also provides a new way for the production of medicinal active ingredients. [0003] Tianshan Saussurea involucrata (Saussurea involucrata Kar.et Kir.), also known as Xinjiang Saussurea involucrata, Saussurea involucrata Kar.et Kir., is a perennial herbaceous plant belonging to the family Asteraceae and is a national medicine with great develop...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/02C07H15/203C07H1/08C07C69/732C07C67/48
Inventor 戴均贵陈日道杨林
Owner INST OF MATERIA MEDICA AN INST OF THE CHINESE ACAD OF MEDICAL SCI
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