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Thiobacillus ferrooxidans with high production of magnetosome and application thereof

A technology of Thiobacillus ferrooxidans and ferrous oxide, applied in the biological field, can solve problems such as scarcity

Active Publication Date: 2019-01-22
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching, there are very few reports on the mass production of magnetosomes by Acidophilus ferrooxidans

Method used

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  • Thiobacillus ferrooxidans with high production of magnetosome and application thereof
  • Thiobacillus ferrooxidans with high production of magnetosome and application thereof
  • Thiobacillus ferrooxidans with high production of magnetosome and application thereof

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specific Embodiment approach

[0027] The technical solution of the present invention is not limited to the specific implementations listed below, but also includes any combination of the specific implementations.

[0028] The invention provides a high-yield magnetosome-producing Thiobacillus ferrooxidans BYM (Acidithiobacillus ferrooxidans BYM, A. ferrooxidans BYM) and its application. A strain of A.ferrooxidansBYM with high magnetosome production was bred by the combination of Race-track liquid magnetophoresis separation and semi-solid plate magnetophoresis separation. The strain has the advantage of being easy to cultivate, the growth period is 24-36 hours, and 6.28 mg of magnetosomes are produced per 1 g of bacteria. The laboratory can use the high-yield magnetosome strain A.ferrooxidans BYM as a biological factory to produce magnetosomes, and the produced magnetosomes can be used for drug carriers and other purposes.

[0029] The following is an introduction through a specific implementation manner. ...

specific Embodiment approach 1

[0032] Acidithiobacillus ferrooxidans (A. ferrooxidans) was isolated from a water sample of a copper mine in Baiyin City, Gansu Province.

[0033] Acidophilus ferrooxidans (A.ferrooxidans) was inoculated in the liquid culture medium, and the liquid culture medium was prepared according to the following ratio: in every 1000ml liquid culture medium, by 2.4g (NH 4 ) 2 SO 4 , 0.1g of KCl, 0.5g of K 2 HPO 4 , 0.5g of MgSO 4 ·7H 2 O, 0.01g of Ca(NO 3 ) 2 , 40g of FeSO 4 and the remainder of distilled water, and adjust the pH to 2.0.

[0034] According to the inoculum amount of 8-12% (v / v), in this embodiment, the acidophilic Thiobacillus ferrooxidans (A. ferrooxidans) is inoculated in the liquid medium. Cultivate on a shaker at 120 rpm at a temperature of 30° C., and cultivate for 24 to 48 hours until the logarithmic phase is ready for use.

specific Embodiment approach 2

[0035] Separation of Thiobacillus ferrooxidans Race-track liquid magnetophoresis:

[0036] ① Take the bacterium liquid cultured to the logarithmic phase in Embodiment 1, and filter it with a microporous membrane (the pore size is 0.22 μm) to obtain acidophilic Thiobacillus ferrooxidans (A.ferrooxidans) thallus;

[0037] ② Use 1mL of H at pH 2 2 SO 4 Rinse the solution and centrifuge at 2000rmp for 5min;

[0038] ③Take the supernatant, centrifuge at 10000rmp for 5min, and take the precipitate;

[0039] ④ Repeat steps ② and ③ 3 times to obtain precipitation;

[0040] ⑤ Suspend with 500 μL of 0K solution, and collect in Pasteur tube for 30 minutes;

[0041] ⑥Take 3 Pasteur tubes, and add the bacterial solution to the end of the Pasteur tubes. After 30 minutes of magnetic collection, take the tip 2cm, suck out the liquid with a 1ml syringe, inject it into the liquid medium, and cultivate it on a shaker at 120rpm at a temperature of 30°C.

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Abstract

The invention relates to thiobacillus ferrooxidans with high production of magnetosome and application thereof. The strain of thiobacillus ferrooxidans has an accession number of CCTCC NO: M 2018630.The strain has the advantages of easy cultivation, the growth cycle is 24 to 36 hours, and 6.28 mg of magnetosomes is produced in per 1 g of thalli of thiobacillus ferrooxidans. The thiobacillus ferrooxidans with high production of magnetosome can be used as a biological plant to produce magnetosomes, and is further used for pharmaceutical carriers and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a high-magnetosome-producing Thiobacillus ferrooxidans and its application. Background technique [0002] Magnetosomes are magnetic nanoparticles with uniform size distribution, non-toxicity, high biological activity, and high saturation magnetization. Compared with ordinary artificial nano-magnetic particles, the biggest feature is that the magnetic particles are encapsulated in biofilms. Based on the various characteristics of magnetosomes, magnetosomes are widely used in biochemistry, clinical medicine, environmental protection and other fields (patent: CN201711325632.5) (patent: CN 201410748312.0). [0003] At present, the vast majority of magnetosomes come from magnetotactic bacteria, but magnetotactic bacteria are generally obligate microaerobic or facultative anaerobic bacteria, which grow slowly and have low growth. It is difficult to simulate their living environ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12R1/01
CPCC12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 晏磊张爽刘涛王伟东王彦杰
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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