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High-density bacterium culturing method and its biological reactor

A technology of cell culture and bioreactor, which is applied in the field of cell high-density circulation or continuous culture method and its bioreactor device, which can solve the problem of difficult to eliminate bacteria pollution, difficult circulation or continuous high-density culture, etc. problems, to achieve the effect of improving equipment utilization, shortening production cycle, and reducing operating costs

Inactive Publication Date: 2006-05-31
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the organic membrane used in the high-density purebred culture method using an external hollow fiber membrane tube cannot be sterilized by heat, so it is difficult to eliminate the contamination of bacteria in practical applications, and it is even more difficult to complete the circulation or continuous high-density cultivation. density culture

Method used

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  • High-density bacterium culturing method and its biological reactor

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Effect test

Embodiment 1

[0024] High-density culture of lactic acid bacteria circulation

[0025] First, add beef extract 1%, peptone 1%, yeast extract 1%, glucose 15%, Tween 80 0.05% pH6.5 medium in the cleaned bioreactor, and bioreactor 2 and tank with the conventional method The pipelines connected to the body are heat sterilized, and the seeds can be inserted into the seed solution after being cooled by the jacket cooling water and cultivated at 30 °C. Before the end of the cultivation, when K2K3K4 is closed, open K1K6K8 and use steam to sterilize the inorganic membrane module, pump 4 and the pipeline connected to it, then close K6K7K8 and cool down to room temperature properly. When the concentration of lactic acid bacteria in the bioreactor reaches 10 5 cfu / ml-10 8 When cfu / ml, turn on K2K1K5, turn on pump 4, circulate and filter through the membrane tube, discard the clear liquid through the clear liquid outlet, and return the concentrated bacterial liquid to the bioreactor. After circulating...

Embodiment 2

[0027] Continuous high-density culture of Escherichia coli

[0028] First, add lactose 0.5%, tryptone 2%, potassium dihydrogen phosphate 0.275%, dipotassium hydrogen phosphate 0.275%, NaCl 0.5%, sodium lauryl sulfate 0.05% pH6.6- 7.0 culture medium, heat-sterilize the bioreactor 2 and the pipeline connected with the tank according to the conventional method, and after cooling through the jacket cooling water, it can be connected to the seed liquid for cultivation under certain conditions. Before the end of the cultivation, when K2K3K4 is closed, open K1K6K8 and use steam to sterilize the inorganic membrane module, pump 4 and the pipeline connected to it, then close K6K7K8 and cool it down properly. When the concentration of E. coli in the bioreactor reaches 10 5 cfu / ml-10 8 When cfu / ml, turn on K2K1K5, turn on pump 4, circulate and filter through the membrane tube, discard the clear liquid through the clear liquid outlet, and return the concentrated bacterial liquid to the bi...

Embodiment 3

[0030] Continuous High Density Culture of Yeast

[0031] First, add wort juice (2%-12%) in the cleaned bioreactor, carry out thermal sterilization to bioreactor 2 and the pipeline that communicates with the tank body according to the conventional method, after cooling by jacket cooling water, It can be cultivated under certain conditions by accessing the seed solution. Before the end of the cultivation, when K2K3K4 is closed, open K1K6K8 and use steam to sterilize the inorganic membrane module, pump 4 and the pipeline connected to it, then close K6K7K8 and cool it properly. When the yeast concentration in the bioreactor reaches 10 5 cfu / ml-10 8 When cfu / ml, turn on K2K1K5, turn on pump 4, circulate and filter through the membrane tube, discard the clear liquid through the clear liquid outlet, and return the concentrated bacterial liquid to the bioreactor. After circulating and filtering, the bacterial cells in the bioreactor After the concentration is increased by 5-10 times...

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Abstract

A culture of high-density cell and its biological reactor are disclosed. It is characterized by installing inorganic membrane filter assembly and feed-supplement pot on biological reactor and connecting the three parts into a biological reactor system for cell circulation high-density culture or high-density continuous culture by control loop. It can combine the cell culture with membrane filter concentrate to obtain the needed metabolic product. It costs low, has short reaction time and higher production efficiency.

Description

technical field [0001] The invention relates to a cell culture method and a biological reaction device thereof, in particular to a cell high-density circulation or continuous culture method and a biological reaction device thereof. Background technique [0002] Membrane bioreactor is a new type of equipment formed by introducing membrane separation technology into bioreactor. The existing membrane bioreactor (MBA) is mainly used for sewage treatment. Its working principle is to use membrane to intercept the activated sludge In order to improve the efficiency of sewage biological treatment by microorganisms, the core problem is to solve the problem of membrane filtration pollution, that is, the decrease of filtration flux in the process of sewage biological treatment. Since the biological sewage treatment is an open fermentation process, it does not involve the pollution of bacteria, that is, the membrane equipment used in this type of bioreactor does not need to consider the...

Claims

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Application Information

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IPC IPC(8): C12N1/00C12N5/00
Inventor 熊晓辉熊强陆利霞韩亦龙
Owner NANJING UNIV OF TECH
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