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Magnetospirillum magneticum as well as preparation method and application thereof

A magnetospiral, formula technology, applied in the direction of botany equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of not easy to form thrombus, no in-depth research and large-scale production and application, and achieve difficult The effect of thrombus formation, simple and efficient design, good biocompatibility and safety

Active Publication Date: 2015-06-17
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patents ZL02115867.3, ZL02115868.1 and CN1952112A respectively proposed "a kind of sludge magnetotactic bacteria and its separation and purification and preparation of magnetosomes", "a kind of iron ore magnetotactic bacteria and its separation and purification and preparation of magnetosomes". method for magnetosomes” and “a facultative anaerobic magnetotactic bacterium and its isolation, pure culture method, and purification and purification method of magnetosomes”, although magnetosomes can be obtained, there is no in-depth research and large-scale production and application reports
The production of magnetosomes with smaller particle diameter, less thrombus formation, and better dispersion performance will have greater application value than traditional magnetosomes. However, the use of magnetotactic bacteria to produce superparamagnetic magnetosomes has not been reported so far.

Method used

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  • Magnetospirillum magneticum as well as preparation method and application thereof
  • Magnetospirillum magneticum as well as preparation method and application thereof
  • Magnetospirillum magneticum as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] A kind of Magnetospira ME-1, obtained by screening through the following steps:

[0057] 1) Collection and capillary separation of magnetotactic bacteria samples:

[0058] (1) Magnetotactic bacteria collection: The collection device is inverted, and the cylindrical plastic collection box 1 is filled with filtered and sterilized in-situ water (the in-situ water is collected from the water sample close to the mud-water interface and sterilized by filtration with a 0.22 μm filter membrane. ), seal the filter paper 2 with a metal tripod 3, install the magnet 4 and the fishing line 5 into the natural water environment, make the device stand at the mud-water interface and the filter paper surface of the collection device is located 1-2 cm above the mud-water interface, and place it for 12 Take it out after 1 hour, remove the tripod, carefully open the filter paper, suck the collection solution in the collection box, and carry out optical microscope observation, electron micro...

Embodiment 2

[0088] Phylogenetic and gene island analysis of Magnetospirillum sp.ME-1 strain

[0089] ⑴ Genome Extraction

[0090] Genomic DNA of strain ME-1 was extracted by glass rod stirring method (refer to Journal of Molecular Biology, 1961, 3: 208-218 for the method). Roche GS FLX shotgun library was built, Roche GS FLX Titanium System454 sequencer was used for sequencing, and 79 contigs were generated after sequence splicing.

[0091] (2) Phylogenetic analysis of the measured 16SrDNA sequence of the strain (the sequence is shown in the sequence table as SEQ ID No. 2) shows that the strain belongs to the genus Magentospirillum, and the 16SrDNA sequence of the strain is similar to Magentospirillum magneticum AMB-1, Magentospirillum magnetotacticum MS-1, and Magentospirillum respectively. The 16SrDNA sequence of gryphiswaldenseMSR-1 has 99.4%, 98.1%, and 96.3% sequence similarity, so it was named Magentospirillum sp.ME-1.

[0092] (3) Using Mauve software, the measured genome was com...

Embodiment 3

[0096] Fermentation of Magnetospirillum sp.ME-1 Strain

[0097] Fermentation culture test of Magnetospirillum sp.ME-1 in 10L fermenter:

[0098] The specific conditions are: add 6L of growth medium to the fermenter, inoculate with 10% inoculum, culture conditions at 30°C, air flow at 20L / h, stirring speed coupled with 2% dissolved oxygen, feed coupled with pH 6.86. Magnetospirillum sp.ME-1 grows to OD in about 40 hours under this culture condition 565 2.5.

[0099] The formula of the growth medium is: vitamin mixture (see Table 2 for formula) 1ml / l, mineral mixture (see Table 3 for formula) 2ml / l, potassium phosphate buffer (0.1M, pH7.0) 10ml / l l, sodium succinate 0.05% (m / V), sodium acetate 0.05% (m / V), yeast extract 0.025% (m / V), ammonium chloride 0.05% (m / V), magnesium sulfate 0.01% ( m / V), ferric citrate 100μM, in which the vitamin mixture and mineral mixture need to be sterilized by the culture medium and then added with filter-sterilized solution separately.

[0100]...

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Abstract

Disclosed are a magnetospirillum magneticum, a preparation method therefor, and a use thereof. In the preset invention, a magnetospirillum magneticum Magnetospirillum sp. ME-1, CCTCC No: M2013260 which can produce superparamagnetic magnetosomes is separated fresh water. The strain is a magnetospirillum magneticum with a size of 2.62~4.83×0.44~0.62μm, and comprises a magnetosome chain consisting of 10 to 31 magnetosome particles. A component of each magnetosome is ferroferric oxide, is a cuboctahedron or an octahedron in shape, and has a size of 26±5nm which is in a superparamagnetic range. The magnetospirillum magneticum strain has a specific magnetosome gene island sequence and can produce magnetosomes with a yield of 177mg / L by directly using air fermental cultivation. The strain has excellent fermentability, and produces magnetosome particles which are superparamagnetic, and has a great industrialization popularization prospect.

Description

technical field [0001] The invention relates to the field of nanobiology technology, in particular to a magnetospiral ME-1 capable of producing superparamagnetic nano-magnetosomes, and also to a device for screening the magnetospiral, and to the magnetotactic The fermentation method of the spirilla also relates to the application of the magnetotactic spirilla in the preparation of superparamagnetic magnetosomes. Background technique [0002] Magnetotactic bacteria are a type of special bacteria that can form nano-magnetosomes in their cells. The magnetosomes formed by them have the characteristics of external magnetic field controllability, stable crystal form, nanometer size, excellent biological safety, and are not easy to be covered by biofilms. Reunion and other characteristics show great advantages over other materials in immune detection, drug delivery, multifunctional molecular carriers, tumor magnetic hyperthermia, nuclear magnetic resonance imaging, etc., especially...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/31C12M1/00C12R1/01
CPCC12N1/20C12P3/00C12N1/205C12R2001/01
Inventor 刘朋明高梅影陈亚军
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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