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Method for producing anoectochilus roxburghii polysaccharose through suspension cultivation of cell

A technology of suspension culture cells and clematis, which is applied in the direction of fermentation, etc., can solve the problems of long production cycle, low yield of target product, low growth rate of biomass, etc., achieve uniform supply, shorten cell culture cycle, and increase product yield rate effect

Active Publication Date: 2015-03-11
WEST ANHUI UNIV
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

The technology of using cell engineering technology to produce clematis polysaccharides has been studied at home and abroad for a long time, and many achievements have been made. However, there are many difficult problems in the production process, such as low biomass growth rate, low yield of target products, and production cycle. long wait
There is no report on the production of Clematis clematis polysaccharides by suspension culture of A.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0016] 1. Training stage

[0017] (1) Single cell preparation

[0018] Take clematis capsules, sterilize them according to the requirements of tissue culture technology, and carry out aseptic sowing. The sowing medium is an improved 1 / 2MS solid medium. Take them out after they germinate into protocorms. Choose protocorms with bright color, large individuals and vigorous vitality. For bulbs, separate and prepare single cells by enzymatic hydrolysis, filter with a microporous filter with a pore size of 80 microns, separate the filtrate through a low-speed centrifuge, and collect the precipitated cells;

[0019] (2) Single cell line culture

[0020] Inoculate the collected single cells into a baffle shake flask containing improved 1 / 2MS liquid medium, the inoculum amount of fresh cells is 25±2g / L, the light intensity is 3000lux, the color temperature is 5000K, shake culture, adjust the pH value 5.6±2, temperature 22±2°C, when the cell density reaches 10000 cells / mL, the culture...

specific Embodiment 2

[0029] 1. Training stage

[0030] (1) Single cell preparation

[0031] Take clematis capsules, sterilize them according to the requirements of tissue culture technology, and carry out aseptic sowing. The sowing medium is an improved 1 / 2MS solid medium. Take them out after they germinate into protocorms. Choose protocorms with bright color, large individuals and vigorous vitality. For bulbs, single cells were separated by enzymatic hydrolysis, filtered through a microporous filter with a pore size of 60 microns, the filtrate was separated by a low-speed centrifuge, and the precipitated cells were collected;

[0032] (2) Single cell line culture

[0033] Inoculate the collected single cells into a baffle shake flask containing improved 1 / 2MS liquid medium, the inoculum amount of fresh cells is 25±2g / L, the light intensity is 2000lux, the color temperature is 6000K, shake culture, adjust the pH value 5.6±2, temperature 22±2°C, when the cell density reaches 20000 cells / mL, the c...

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Abstract

A method for producing anoectochilus roxburghii polysaccharose through suspension cultivation of cells. In the method, the anoectochilus roxburghii polysaccharose is produced through a cell cultivation reactor for culturing anoectochilus roxburghii cells. The method includes following steps: separating undifferentiated single cells from a high-activity protocorm through an enzymolysis method, wherein the protocorm is formed through sterile seeding and germinating of seeds of anoectochilus roxburghii; further performing cultivation to obtain a single cell line; and finally performing cell mass amplification cultivation. In the amplification cultivation, a plant growth regulator and a biological enzyme preparation are added to a culture medium so that cell merisis can be activated and cell aggregation agglomeration growth can be prevented. Meanwhile, proper illumination intensity and color temperature are selected for regulating expression of target products. The method can greatly reduce a cultivation period of the cells, can increase yield of the anoectochilus roxburghii polysaccharose, is simple in processes, is stable and is suitable for industrialized production.

Description

technical field [0001] The invention relates to sugar production technology, in particular to plant polysaccharide production technology, in particular to a method for producing clematis polysaccharide by adopting a cell culture method. Background technique [0002] Clematis clematis polysaccharides are one of the main components of clematis clematis, which can treat bronchitis, nephritis, cystitis, diabetes, hematuria, rheumatoid arthritis, acute and chronic liver disease, hypertension, arteriosclerosis, and cerebral thrombosis. etc. have auxiliary effects. The pharmacological effects of clematis polysaccharides are mainly manifested in the functions of nutrition, anti-aging, and regulating the immunity of the human body. With the development of society, people pay more and more attention to health, and its demand will also increase. However, the traditional method of extracting clematis polysaccharides is to use the roots, stems, leaves and other plant bodies of clematis ...

Claims

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Application Information

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IPC IPC(8): C12P19/04
Inventor 邓辉陈乃富刘文中陈存武韩邦兴
Owner WEST ANHUI UNIV
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