Method for producing Dendrobium huoshanense alkaloids by cell suspension culture

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

Active Publication Date: 2016-08-31
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 Dendrobium alkaloids has been studied at home and abroad, and many achievements have been made. However, there are many difficult problems in the production process, such as complex process, low biomass growth rate, and low yield of target products. , long production cycle, etc.
There is no report on the production of Dendrobium alkaloids in Dendrobium Huoshanense by suspension culture in an air-lift stirred tank, so it is very meaningful to develop a method for efficiently producing alkaloid natural products in Dendrobium Huoshanense

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] This embodiment discloses a method for producing Dendrobium alkaloids 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 is 23g / L, the light...

Embodiment 2

[0041] 1. Training stage

[0042] (1) Single cell preparation

[0043] 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;

[0044] (2) Single cell line culture

[0045] 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

[0055] 1. Training stage

[0056] (1) Single cell preparation

[0057] 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;

[0058] (2) Single cell line culture

[0059] 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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PUM

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Abstract

The invention discloses a method for producing Dendrobium huoshanense alkaloids by cell suspension culture, particularly a method for producing Dendrobium huoshanense alkaloids by performing suspension culture on Dendrobium huoshanense cells by using a cell culture reactor. The method comprises the following steps: (1) unicell preparation; (2) unicell line culture; (3) cell population amplification culture; and (4) Dendrobium huoshanense alkaloid extraction: when the cell density reaches 60000-80000 / ml, collecting the Dendrobium huoshanense cells, lowering the culture temperature to 7-10 DEG C, continuing culturing for 5-7 days, and extracting the Dendrobium huoshanense alkaloids. The method can greatly shorten the cell culture period and enhance the Dendrobium huoshanense alkaloid product yield; and the technique is simple and stable, and is 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 Dendrobium Huoshanense alkaloids by suspension culture cells. Background technique [0002] Dendrobium Huoshanense alkaloids are one of the main components of Dendrobium Huoshanense. The pharmacological effects of alkaloids are mainly manifested in anti-tumor, cardiovascular, gastrointestinal inhibition, pain relief and antipyretic effects. With the development of society, people pay more and more attention to health, and their demand will also increase. However, the traditional method of extracting the alkaloids of Dendrobium Huoshanense is to extract them from the roots, stems, leaves and other plants of Dendrobium Huoshanense , the products obtained by this process can no longer meet the future market demand for intensive processing of Dendrobium huoshanense. [0003] With the development of plant cell engineering technology, the use of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D491/08C12N5/04C12N5/02
CPCC07D491/08C12N5/04C12N2500/10C12N2501/73C12N2501/998C12N2501/999
Inventor 邓辉陈乃富陈存武韩邦兴何晓梅
Owner WEST ANHUI UNIV
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