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Method for producing dendrobium huoshanense polysaccharide through suspension culture cells

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: 2016-08-17
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 polysaccharides 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 growth rate of biomass, low yield of target product, Long production cycle, etc.
There is no report on the production of Dendrobium Huoshanense polysaccharides by suspension culture of Dendrobium Huoshanense in an air-lift stirred tank, so it is very meaningful to develop a method for efficiently producing natural products of Dendrobium Huoshanense polysaccharides

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0022] Preparation of reference solution: take an appropriate amount of anhydrous glucose reference substance dried at 105°C to constant weight, accurately weigh, dissolve in water and quantitatively prepare a solution containing 100 μg per 1 ml.

[0023] Drawing of standard curve: draw 0.1, 0.2, 0.3, 0.4, 0.5ml of glucose reference solution (concentration: 0.1g / L) into a 10mL test tube, add distilled water to 1.0ml, add 1ml of 5% phenol solution, shake well , add 5ml of sulfuric acid, shake well, heat in a boiling water bath for 20min, take it out, put it in an ice bath to cool for 5min, take the corresponding reagent as a blank, measure the absorbance at the wavelength of 488nm, take the absorbance as the ordinate and the concentration as the horizontal Coordinates, draw a standard curve. Taking the absorbance A as the ordinate and the glucose concentration C as the abscissa, as a control curve, the regression equation was A=0.0367C+0.0412, r=0.9998, and the linear range was...

Embodiment 1

[0026] This embodiment discloses a method for producing Dendrobium huoshanense polysaccharide by suspension culture cells, comprising the following steps:

[0027] 1. Cultivation stage

[0028] (1) Single cell preparation

[0029] Take Dendrobium huoshanense capsules, sterilize them according to the technical requirements of tissue culture, and then perform aseptic sowing. The seeding medium is solid 1 / 2 MS medium. After germination into protocorms, take them out. Choose protocorms with bright colors, large individuals, and vigorous vitality. Enzymatic hydrolysis is used to separate and prepare single cells, which are filtered with a microporous filter with a pore size of 80 microns, the filtrate is separated by a low-speed centrifuge, and the precipitated cells are collected;

[0030] (2) Single cell line culture

[0031] The collected single cells were inoculated into the baffle shake flask containing the first MS liquid medium. The single cell inoculation volume was 23 g / ...

Embodiment 2

[0042] 1. Cultivation stage

[0043] (1) Single cell preparation

[0044] Take Dendrobium huoshanense capsules, sterilize them according to the technical requirements of tissue culture, and then perform aseptic sowing. The seeding medium is solid 1 / 2 MS medium. After germination into protocorms, take them out. Choose protocorms with bright colors, large individuals, and vigorous vitality. Enzymatic hydrolysis is used to separate and prepare single cells, which are filtered with a microporous filter with a pore size of 80 microns, the filtrate is separated by a low-speed centrifuge, and the precipitated cells are collected;

[0045] (2) Single cell line culture

[0046] The collected single cells were inoculated into a baffled shake flask containing the first MS liquid medium. The single cell inoculation volume was 25 g / L, the light intensity was 3000 lux, the color temperature was 5000 K, the culture was shaken, the pH value was adjusted to 5.8, and the temperature was adjust...

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Abstract

The invention discloses a method for producing dendrobium huoshanense polysaccharide through suspension culture cells. According to the method, dendrobium huoshanense polysaccharide is produced by conducting suspension culture on the dendrobium huoshanense cells through a cell culture reactor. The method comprises the following steps of 1, single-cell preparation; 2, culture of a single-cell system; 3, amplification culture of a cell population; 4, extraction of dendrobium huoshanense polysaccharide, wherein when the cell density reaches 60000-80000 cells / ml, the dendrobium huoshanense cells are collected, and dendrobium huoshanense polysaccharide is extracted. By means of the method, the cell culture cycle can be remarkably shorted, the yield of dendrobium huoshanense polysaccharide products is raised, the process is simple and stable, and the method is suitable for industrialized production.

Description

technical field [0001] The invention relates to the technical field of Dendrobium huoshanense, in particular to a method for producing Dendrobium huoshanense polysaccharide by suspension culture cells. Background technique [0002] Dendrobium huoshanense polysaccharide is one of the main components of Dendrobium huoshanense. The pharmacological effects of Dendrobium huoshanense polysaccharide are mainly manifested in immunity, strengthening, anti-aging, anti-tumor and other effects. With the development of society, people pay more and more attention to health, and its demand will also increase. However, the traditional extraction method of Dendrobium huoshanensis polysaccharide is extracted 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 Huoshan. [0003] With the development of plant cell engineering technology, the use of cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12N5/02C12N5/04
CPCC12N5/04C12P19/04
Inventor 邓辉陈乃富陈存武韩邦兴王广林
Owner WEST ANHUI UNIV
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