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Method for black moss cell high-density culture of stable high-yield black moss polysaccharide

A technology of high-density cultivation and growing vegetables, which is applied in the cultivation of microalgae cells and in the field of biology, and can solve the problems of low cell density, high cost, and long cultivation process

Inactive Publication Date: 2007-10-31
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the cell culture of hair vegetables reported in the literature is all using photoautotrophic culture. This culture method has the disadvantages of low cell density, dependence on light intensity, long culture process and high cost. Therefore, heterotrophic culture and mixed nutrient culture are the best ways to achieve high density effective way to cultivate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Shake flask mixed nutrient culture Nostocella cells

[0022] 1. Preservation and pre-cultivation of Nostocia spp. cell species

[0023] The culture medium used in the embodiment of the present invention to preserve the Nostocella cell species consists of: sodium bicarbonate 1.2%, sodium nitrate 0.12%, ammonium nitrate 0.03%, adding dipotassium hydrogen phosphate, calcium chloride, magnesium sulfate, copper sulfate, sulfuric acid Ferrous, zinc sulfate, sodium molybdate, boric acid, cobalt chloride, pH 8.5, stored at 8°C, 500lux under low light, transferred once a month. Before carrying out formal cultivation, all will pre-cultivate the Nostocella cell species, so that it transitions from a slow growth state to a fast growth state, and reaches a certain cell concentration. The pre-culture liquid medium used is the basal medium supplemented with glucose, according to OD 750 Inoculate about 0.05% of the inoculum into the pre-culture medium, culture staticall...

Embodiment 2

[0028] Example 2: Bioreactor Mixed Nutrition Cultivation of Nostocella cells

[0029] 1. Preservation and pre-cultivation (same as Example 1) of Nostocella cell species.

[0030] 2. Bioreactor Culture of Nostocella cells

[0031]Get the pre-cultivated seed solution, inoculate it into a mixed nutrient medium with a glucose concentration of 2.0-4.5 g / L and a sodium nitrate concentration of 1.0-4.5 g / L with an inoculum size of 5-10% (v / v), and form the The medium was added to a 20L photobioreactor, and tap water was added to 15L to sterilize it. When the temperature dropped to about 25°C, it was inoculated with Nostocella cell seed according to the volume ratio of 10%, and the cultivation was started. Culture conditions: light intensity 40~120μmol / m 2 .s, the pH is controlled above 7.0, the culture temperature is 20-30°C, and the air flow rate is 0.06-0.1vvm; when the growth of Nostocella cells enters the late stage of the stable period, the culture is terminated, and the cells...

Embodiment 3

[0032] Embodiment 3: 5L closed bioreactor heterotrophic culture of Nostocella cells

[0033] After pre-cultivating the Nostocella cells, they were inserted into a 5L closed bioreactor with a 10% inoculation amount. The culture conditions were: no light, and other conditions were the same as in Example 2.

[0034] Advantage and positive effect of the present invention are:

[0035] The high-density culture process established by the present invention adopts mixed nutrition or heterotrophic culture mode, which breaks through the traditional photoautotrophic nutrition mode of Nostocella cell growth, reduces the dependence on light in the culture process, and adds organic carbon sources to greatly improve The growth rate of the Nostoc sativa cells is improved, the culture period of the Nostoc sativa cells is shortened, and the yield of the Nostoc sativa polysaccharides can be improved at the same time. By using the method of the present invention to cultivate the cells of Nostocc...

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Abstract

The invention discloses a high density culture method of stable and high output hair weed cell, which comprises the following steps: pre-culturing hair weed cell seed; adjusting the initial pH value of the culture medium in shake or biological reactor above 7. 0; switching in pre-culturing hair weed cell seed in the culture medium; ending the culture when the growth of hair weed cell enter late time of the stable period; collecting the hair weed cell and culture liquid. This invention adopts mixed nourishing or heterotrophy culture model, which lays the groundwork for endurance utilization of hair weed source.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the field of culture of microalgae cells, in particular to a method for high-density cultivation of Nostocchi sativa cells with stable and high yield polysaccharides. Background technique [0002] Nostoc flagelliforme belongs to Prokaryote (Pro-caryotae), Cyanophyta (Cyanophta), Cyanophyceae (Cyanophceae), Hormogonales, Nostocaceae, Nostoc ( Nostoc) is a terrestrial economic cyanobacteria distributed in the arid regions of northern and northwestern my country. Because of the special ecological environment requirements and geographical distribution of Nostocs, and its natural growth is very slow, the unreasonable harvesting of Nostomas will easily lead to the reduction of wild Nostomas resources, which will cause serious damage to the vegetation in the place where Nostilatis grows, resulting in The land is desertified and the ecological environment is deteriorating day by day. Since July 20...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12P19/04
Inventor 贾士儒于海峰苏建宇林永贤
Owner TIANJIN UNIV OF SCI & TECH
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