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Optimal dissolved oxygen control method for producing magnetosome by cultivating magnetotactic bacteria

A control method and technology of magnetosomes, applied in bacteria and other directions, can solve the problems that the speed cannot keep up with the increase rate of cells, the decline of magnetosome volume, and the decline of magnetosome yield, etc., to achieve fast cell growth and cell yield. High, easy-to-use effects

Inactive Publication Date: 2009-05-20
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] As the base number of cells increases, the growth of cells per unit time increases rapidly, while the synthesis of magnetosomes in a single cell is relatively stable, and the maturation of magnetosomes takes a long time, so magnetosomes The speed of synthesis will not keep up with the speed of cell growth, and the speed of iron absorption and utilization by cells will not keep up with the speed of cell growth, so that the number of magnetosomes in a single cell will decrease (see figure 2 , 3 ), which will lead to a decrease in the amount of magnetosomes produced by consuming a unit weight of nutrient substrate, that is, a decrease in the yield of magnetosomes

Method used

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  • Optimal dissolved oxygen control method for producing magnetosome by cultivating magnetotactic bacteria
  • Optimal dissolved oxygen control method for producing magnetosome by cultivating magnetotactic bacteria
  • Optimal dissolved oxygen control method for producing magnetosome by cultivating magnetotactic bacteria

Examples

Experimental program
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Embodiment 1

[0041] Example 1 The method of cultivating magnetobacteria to produce magnetosomes

[0042] (1) In the culture medium containing 0.03g / L sodium lactate, 0.05g / L ammonium chloride and 0g / L ferric citrate, shake culture Magnetospira MSR-1, obtain seed culture solution, wherein said culture medium initial pH The value is 6.5;

[0043] (2) Add K-free in 42L automatic fermenter 2 HPO 4 The above culture medium was sterilized, continuously fed with sterile air (0.3L / min), and continuously stirred at 100rpm. When the temperature of the culture medium dropped below 40°C, add pre-sterilized K 2 HPO 4 The final concentration is 0.1g / L. When the temperature of the culture medium drops to 35°C, inoculate the seed solution activated in step (1) into the tank, and adjust and control the pH to 6.5 with hydrochloric acid;

[0044] (3) During the culture process, samples were taken every 2 hours to detect the cell density and lactic acid content and the iron content of the culture solutio...

Embodiment 2

[0047] Example 2 The method of cultivating magnetobacteria to produce magnetosomes

[0048] (1) In the culture medium that contains 10.4g / L sodium lactate, 3.2g / L ammonium chloride and 2.08mg / L iron citrate, shake and cultivate Magnetospira MSR-1, obtain seed culture solution, wherein said culture medium initially The pH value is 7.5;

[0049] (2) Add K-free in 42L automatic fermenter 2 HPO 4 After the above-mentioned culture medium was sterilized, the sterile air (1.5L / min) was continuously fed in, and the stirring was continued at 200rpm. When the temperature of the culture medium dropped below 40°C, pre-sterilized K 2 HPO 4 The final concentration is 3g / L. When the temperature of the culture medium drops to 25°C, inoculate the seed solution activated in step (1) into the tank, and adjust and control the pH to 7.5 with lactic acid;

[0050] (3) During the culture process, samples were taken every 2 hours to detect the cell density and lactic acid content and the iron co...

Embodiment 3

[0053] Example 3 The method of cultivating magnetobacteria to produce magnetosomes

[0054] (1) In the culture medium that contains 2.6g / L sodium lactate, 0.61g / L ammonium chloride and 13mg / L iron citrate, shake and cultivate Magnetospira MSR-1, obtain seed culture solution, wherein said culture medium initial pH Value is 7.0;

[0055] (2) Add K-free in 42L automatic fermenter 2 HPO 4 The above culture medium was sterilized, continuously fed with sterile air (0.3L / min), and continuously stirred at 100rpm. When the temperature of the culture medium dropped below 40°C, add pre-sterilized K 2 HPO 4 The final concentration is 0.5g / L. When the temperature of the culture medium drops to 28°C, inoculate the seed solution activated in step (1) into the tank, and adjust and control the pH to 7.0 with hydrochloric acid;

[0056] (3) During the culture process, samples were taken every 2 hours to detect the cell density and lactic acid content and the iron content of the culture sol...

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Abstract

The invention provides an optimal dissolved oxygen controlling method for producing magnetosome through magnetic bacteria cultivation, under the precondition ensuring that cell growth is not limited by nutrition, the method takes the speed slowing of cell growth as a signal that dissolved oxygen reaches and gets lower than the critical dissolved oxygen of cell growth, and the method controls rotation speed and aeration flow speed so as to maintain the dissolved oxygen constantly at the critical dissolved oxygen of cell growth, which is the optimal dissolved oxygen level that is suitable for both cell growth and magnetosome synthesis. Experiments prove that by adopting the optimal dissolved oxygen controlling method for producing magnetosome through magnetic bacteria cultivation, not only the final cell cultivation level and the magnetosome yield, but also the cell growth speed and the magnetosome throughput rate achieve the highest level in existing reports.

Description

technical field [0001] The invention relates to the field of bacterial culture, in particular to an optimal dissolved oxygen control method for cultivating magnetobacteria to produce magnetosomes. Background technique [0002] Magnetotactic bacteria are a class of Gram-negative bacteria with magnetotropism, which can synthesize nano-magnetic particles that are arranged in chains and coated with lipid membranes—magnetosomes. Magnetosomes have the main properties and characteristics of excellent carriers, and have broad application prospects. However, it is limited to the laboratory level and has not yet been applied commercially. The main reason is that magnetotactic bacteria are difficult to cultivate artificially, and the synthesis conditions of magnetosomes are not easy to control. The reasons are: 1) magnetotactic bacteria have strict requirements on the nutrition in the environment, and can only use a few kinds of organic acids as carbon sources; 2) in nature Magnetota...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 李颖孙建波姜伟田杰生王珍芳李季伦
Owner CHINA AGRI UNIV
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