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Method for biosynthesizing abietane diterpenoids by culturing cells from cloverleaf

A technique for rosinane diterpenes and culturing cells, which is applied in the field of inducing the biosynthesis of rosin alkane diterpenoids by liquid suspension culture cells of Tricuspid chinensis, can solve the problems of difficulty in obtaining, restricting research and application, low content and the like, and achieves improved purification. Efficiency, cost reduction, easy separation and purification

Inactive Publication Date: 2011-12-28
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, such compounds are mainly extracted from plants, and there are few reports on their chemical synthesis, and they are still in the stage of laboratory research.
Due to the low content of these compounds in plants, it is difficult to obtain them, which greatly limits their further research and application.

Method used

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  • Method for biosynthesizing abietane diterpenoids by culturing cells from cloverleaf
  • Method for biosynthesizing abietane diterpenoids by culturing cells from cloverleaf
  • Method for biosynthesizing abietane diterpenoids by culturing cells from cloverleaf

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Experimental program
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Effect test

Embodiment 1

[0038] (1) Cultivation of the cloverleaf cells:

[0039] Cells were cultured in suspension in liquid suspension:

[0040] The cell strain in the present invention is initially obtained from the suspension culture of the callus induced by the young stem of C. fortunei (C. fortunei), and the Murashige-Skoog (MS) medium is used, and the added concentration is 2 mg / L 2,4-dichloro Phenoxyacetic acid, 0.1mg / L kinetin and 30g / L sucrose, the pH value of the medium is 5.8. 7.5 grams of wet cells obtained by filtration were inoculated into a 500 ml shake flask containing 100 ml of the above medium, and then cultured on a rotary shaker at 100 rpm at 25° C. in the dark.

[0041] (2) Combined induced synthesis of abietane diterpenes:

[0042] On the 7th day of cell growth, methyl jasmonate (MJA) (final concentration in the medium was 300 μM / L) and XAD-7HP resin (100 g / L) were added to the medium, and the culture was continued until the 14th day, respectively. The cells and XAD-7HP were ...

Embodiment 2

[0097] The culture conditions and separation and purification conditions are the same as in Example 1, except that the difference from Example 1 is that MJA (final concentration in the medium is 150 μM / L) and XAD-7HP (100 g / L), continue to cultivate to 14 days, the results are as follows: compound 1 (5.6mg), compound 2 (15.5mg), compound 3 (17.4mg), compound 4 (5.5mg), compound 5 (220mg), compound 6 ( 5mg), Compound 7 (13mg)

Embodiment 3

[0099] The culture conditions and separation and purification conditions are the same as in Example 1, except that the difference from Example 1 is that jasmonic acid (final concentration in the medium is 150 μM / L) and XAD-7HP (100 g / L), continue to cultivate to 14 days, the results are as follows: compound 1 (7.5mg), compound 2 (16mg), compound 3 (20mg), compound 4 (3mg), compound 5 (190mg), compound 6 (6mg), Compound 7 (11.7 mg)

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Abstract

The invention relates to a plant cell culture technology, in particular to a method for biosynthesizing abietane diterpenoid compounds by using cephalotaxus fortune culture cells. The method comprises the following steps of: placing cephalotaxus fortune cells in a common culture medium, and culturing, wherein50 to 150g of the cells are inoculated in each liter of wet weight of the culture medium; and adding an inducer at the final concentration of 10 to 2,000 mu M and 50 to 200g of adsorbent into each liter of culture medium at the initial stage of cell exponential growth, wherein the inducer is an abiological inducer or fungal elicitor. The method is a biosynthesis method and is stable and environment-friendly; and products are completely adsorbed onto the adsorbent, so that the operation process of separation and purification is greatly simplified, the cost is reduced and industrial production is easy to realize. Meanwhile, seven abietane diterpenoid compounds comprising a new compound are synthesized. The abietane diterpenoid compounds have high bioactivity, and are important lead compounds, and the technology has important commercial application prospect.

Description

technical field [0001] The invention relates to a plant cell culture technique, in particular to a method for inducing a liquid suspension culture cell of the cloverleaf to biosynthesize an abietane diterpenoid compound, and a new compound is included in the product at the same time. Background technique [0002] Abietin diterpenoids belong to tricyclic diterpenoids and are an important class of natural compounds. The main reason why this kind of compounds attract people's research interest is that many substances in them have good activity, and many folk medicines contain such substances. For example, hinokiol (compound 2) in the present invention has inhibitory effects on Staphylococcus aureus, Streptococcus, Escherichia coli, Pseudomonas aeruginosa, etc., and is a good antibacterial lead compound. However, these compounds are mainly extracted from plants at present, and there are few reports on their chemical synthesis, and they are still in the stage of laboratory resea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C49/245C07C45/00C07C39/17C07C37/00C07C49/747C07C35/42C07C29/00C07C49/755C12N5/04
Inventor 张卫徐晓辉
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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