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Method for utilizing suspension cultivation cells to produce Dendrobium huoshanense flavone

A technology of suspension culture cells and Dendrobium Huoshanense, which is applied in biochemical equipment and methods, plant cells, tissue culture, etc., can solve the problems of low yield of target products, low growth rate of biomass, long production cycle, etc., and shorten the production cycle of cells The effect of cultivating cycle, uniform supply and improving product yield

Inactive Publication Date: 2016-08-10
WEST ANHUI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The technology of using cell engineering technology to produce flavonoids of Dendrobium huoshanense has been studied at home and abroad for a long time, and many achievements have been made, but there are many problems in the production process, such as complex process, low biomass growth rate, low yield of target product, Long production cycle, etc.
There is no report on the production of flavonoids of Dendrobium huoshanense by suspension culture of dendrobium huoshanense in air-lift stirred tank, so it is very meaningful to develop a method for the efficient production of flavonoids natural products of dendrobium huoshanense

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] This embodiment discloses a method for producing flavonoids from Dendrobium Huoshanense by suspension culture cells, comprising the following steps:

[0025] 1. Training stage

[0026] (1) Single cell preparation

[0027] The capsules of Dendrobium Huoshanense were taken out, sterilized according to the requirements of tissue culture technology, and then aseptically sowed. The sowing medium was a solid 1 / 2MS medium. After the protocorms germinated, they were taken out, and the protocorms with bright color, large individuals and vigorous vitality were selected. Separation and preparation of single cells by enzymatic hydrolysis, filtration with a microporous filter with a pore size of 80 microns, separation of the filtrate by a low-speed centrifuge, and collection of precipitated cells;

[0028] (2) Single cell line culture

[0029] Inoculate the collected single cells into a baffle shake flask containing the first MS liquid medium, the inoculum amount of single cells i...

Embodiment 2

[0038] 1. Training stage

[0039] (1) Single cell preparation

[0040] The capsules of Dendrobium Huoshanense were taken out, sterilized according to the requirements of tissue culture technology, and then aseptically sowed. The sowing medium was a solid 1 / 2MS medium. After the protocorms germinated, they were taken out, and the protocorms with bright color, large individuals and vigorous vitality were selected. Separation and preparation of single cells by enzymatic hydrolysis, filtration with a microporous filter with a pore size of 80 microns, separation of the filtrate by a low-speed centrifuge, and collection of precipitated cells;

[0041] (2) Single cell line culture

[0042] Inoculate the collected single cells into a baffle shake flask containing the first MS liquid medium, the inoculum amount of single cells is 25g / L, the light intensity is 3000lux, the color temperature is 5000K, shake culture, adjust the pH value to 5.8, and the temperature At 22°C, when the cell...

Embodiment 3

[0049] 1. Training stage

[0050] (1) Single cell preparation

[0051] The capsules of Dendrobium Huoshanense were taken out, sterilized according to the requirements of tissue culture technology, and then aseptically sowed. The sowing medium was a solid 1 / 2MS medium. After the protocorms germinated, they were taken out, and the protocorms with bright color, large individuals and vigorous vitality were selected. Separation and preparation of single cells by enzymatic hydrolysis, filtration with a microporous filter with a pore size of 80 microns, separation of the filtrate by a low-speed centrifuge, and collection of precipitated cells;

[0052] (2) Single cell line culture

[0053] Inoculate the collected single cells into a baffle shake flask containing the first MS liquid medium, the inoculum amount of single cells is 27g / L, the light intensity is 4000lux, the color temperature is 6000K, shake culture, adjust the pH value to 6.2, and the temperature At 24°C, when the cell...

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Abstract

The invention discloses a method for utilizing suspension cultivation cells to produce Dendrobium huoshanense flavone. The method is the one for producing Dendrobium huoshanense flavone by utilizing a cell cultivation reactor for suspension cultivation of Dendrobium huoshanense cells, and comprises the following steps: (1) single cell preparation, (2) single cell system cultivation, (3) cell mass scale-up cultivation, and (4) extraction of Dendrobium huoshanense flavone: when the cell density realizes 60,000-80,000 cells per ml, the cultivation temperature is lowered to 7-10 DEG C, cultivation is continued for 5-7 days. Dendrobium huoshanense cells are collected and Dendrobium huoshanense flavone is extracted. The method can greatly shorten the cell cultivation cycle and improve the yield of Dendrobium huoshanense flavone products, and is simple and stable in technology, so as to be suitable for industrial production.

Description

technical field [0001] The invention relates to the technical field of Dendrobium Huoshanense, in particular to a method for producing flavonoids from Dendrobium Huoshanense by suspension culture cells. Background technique [0002] Flavonoids of Dendrobium Huoshanense are one of the main components of Dendrobium Huoshanense. Flavonoids generally refer to a series of compounds formed by connecting two benzene rings (A- and B-rings) with phenolic hydroxyl groups through the central three carbon atoms. The mother nucleus is 2-phenylchromone. Functional groups such as phenolic hydroxyl group, methoxyl group, methyl group and isopentenyl group are often connected in the structure of flavonoids. In addition, it is often combined with sugars to form glycosides. Dendrobium Huoshanense flavonoids have cardiovascular system activity, antibacterial and antiviral activity, antitumor activity, antioxidant free radical activity, anti-inflammatory, analgesic activity, liver protection a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/06C12N5/02C12N5/04
CPCC12N5/04C12P17/06
Inventor 邓辉陈乃富陈存武韩邦兴孙传伯
Owner WEST ANHUI UNIV
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