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Supercritical carbon dioxide method for extraction separation of magnetosomes from magnetotactic bacteria

A technology of carbon dioxide and magnetotactic bacteria, applied in the biological field, can solve problems such as poor stability, poor monodispersity, and low yield, and achieve the effect of overcoming long extraction cycle, good monodispersity, and large particle size

Inactive Publication Date: 2013-12-18
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above-mentioned methods cannot remove some immunogenic substances, and the outer membrane of the magnetic particles extracted by breaking the wall with lysozyme and NaOH has been destroyed, which is easy to cause agglomeration; Hitting may cause the binding protein on the magnetosome membrane to fall off (see Fu Gang, Jiang Wei, Li Ying, etc., Electron Microscopy Observation and Purification of Magnetospira Magnetosome Formation, China Journal of Modern Medicine.2004, 14(5 ):45–49)
Magnetosomes extracted by traditional biological methods have problems such as long production cycle, poor monodispersity, poor stability, difficult removal of residual impurities, and low yield.

Method used

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  • Supercritical carbon dioxide method for extraction separation of magnetosomes from magnetotactic bacteria
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  • Supercritical carbon dioxide method for extraction separation of magnetosomes from magnetotactic bacteria

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Effect test

Embodiment 1

[0026] (1) Organic solvent pretreatment: The magnetotactic bacteria culture medium cultured to the stable phase was separated in a centrifuge (4000 rmp, 20 min), the supernatant was discarded, the bacteria were collected, and the bacteria cells were transferred to 50 mL In the beaker, dissolve the bacterial cells with 0.3 ml of a mixed solvent of chloroform and methanol with a volume ratio of 2:1, shake for 5 min, and the color of the sample changes from pale yellow to milky white.

[0027] (2) Supercritical CO 2 Extraction and removal of organic solvent: add 5mL of redistilled water in the above-mentioned beaker, magnetically stir it to form a suspension, add the above-mentioned suspension to the supercritical carbon dioxide reactor, and put the reactor into a constant temperature of a certain temperature. Stable for a while in a water bath. Pour carbon dioxide gas into the reaction kettle, stir and extract for one hour at a temperature of 35 °C and a pressure of 10 Mpa in a...

Embodiment 2

[0030] (1) Organic solvent pretreatment: The magnetotactic bacteria culture medium cultured to the stable phase was separated in a centrifuge (4000 rmp, 20 min), the supernatant was discarded, the bacteria were collected, and the bacteria cells were transferred to 50 mL In the beaker, dissolve the bacterial cells with 0.3 ml of a mixed solvent of chloroform and methanol with a volume ratio of 2:1, shake for 5 min, and the color of the sample changes from pale yellow to milky white.

[0031] (2) Supercritical CO 2 Extraction and removal of organic solvent: add 5mL of redistilled water in the above-mentioned beaker, magnetically stir it to form a suspension, add the above-mentioned suspension to the supercritical carbon dioxide reactor, and put the reactor into a constant temperature of a certain temperature. Stable for a while in a water bath. Pour carbon dioxide gas into the reaction kettle, stir and extract for one hour at a temperature of 40 °C and a pressure of 25 Mpa in a...

Embodiment 3

[0034] (1) Organic solvent pretreatment: separate the magnetotactic bacteria culture solution cultivated to the stationary phase in a centrifuge (4000 rpm, 20 min), discard the supernatant, collect the bacteria, and transfer the bacteria cells to 50 mL In a beaker, dissolve the bacterial cells with 0.3ml of a mixed solvent of chloroform and methanol with a volume ratio of 2:1, shake well for 5 minutes, and the color of the sample changes from light yellow to milky white.

[0035] (2) Supercritical CO 2 Extraction and removal of organic solvents: add 5mL double-distilled water to the above beaker, stir magnetically to form a suspension, add the above suspension to a supercritical carbon dioxide reactor, and put the reactor into a constant temperature constant temperature Stabilize in a water bath for a period of time. Introduce carbon dioxide gas into the reaction kettle, under the conditions of a certain temperature of the reaction kettle of 60°C and a pressure of 20Mpa, stir...

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Abstract

The invention provides a supercritical carbon dioxide method for extraction separation of magnetosomes from magnetotactic bacteria. The method mainly includes the processes of organic solvent pretreatment, supercritical carbon dioxide extraction separation, organic solvent and water soluble residue removal. By using the method, magnetosomes can be extracted and separated from magnetotactic bacteria under optimal supercritical carbon dioxide fluid temperature, pressure conditions and extraction times. The magnetosomes obtained by extraction separation have the characteristics of small particle size, good monodispersity, high stability, no organic solvent residue, high saturation magnetization intensity, and high extraction separation efficiency, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, and more particularly, to a new method for extracting nanometer magnetosomes from magnetotactic bacteria. Background technique [0002] Magnetotactic bacteria are a class of aquatic bacteria that can move or arrange directionally along the earth's magnetic field or the direction of an external magnetic field. Such bacteria are able to synthesize magnetic particles with biofilm-coated, single-domain-scale crystals called magnetosomes. Magnetosomes have the characteristics of pure composition, unique shape, small and uniform, and outer membrane coating. Due to its huge surface area to volume ratio, no cytotoxicity, not easy to agglomerate and unique biological properties, the nano-scale magnetic particles have broad application prospects in bioengineering, magnetic memory materials, chemical industry, medicine and health, etc. . [0003] There are few literatures about the extraction and purificatio...

Claims

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Application Information

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IPC IPC(8): C01G49/08C01G49/12
CPCY02P20/54
Inventor 安学勤马四红
Owner EAST CHINA UNIV OF SCI & TECH
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