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Culture method of magnetotactic bacteria AMB-1

A technology of magnetotactic bacteria and culturing method, applied in the direction of bacteria, etc., can solve the problems of harsh culture conditions of AMB-1, the relationship is not clear enough, the number of magnetotactic strains is small, and the effect of improving the density of bacteria and the yield of magnetosomes is achieved.

Inactive Publication Date: 2017-02-22
HUAQIAO UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, due to many factors such as the harsh culture conditions of AMB-1, the low level of artificial culture, the culture process is easily contaminated, and the number of isolated and purified magnetotactic strains is small, it is difficult to cultivate large-scale and obtain a large number of magnetosomes. its applied research
Secondly, the relationship between cell density and magnetosome production is not clear enough, and long-term culture leads to a decrease in the amount of synthetic magnetosomes

Method used

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  • Culture method of magnetotactic bacteria AMB-1
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  • Culture method of magnetotactic bacteria AMB-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: the activation of AMB-1 thalline

[0025] (1) The original strain AMB-1 (purchased from the American Type Culture Collection (ATCC)) was activated, and the components of the liquid medium (1653 MSGM) used were prepared according to the ATCC official website. The method of activation and cultivation is as follows: quickly take out the original strain from the -80°C freezer, and immediately place it in a 37°C water bath until the strain changes completely from solid to liquid and no ice-like solids are visible. Immediately inoculate the bacteria into a 10ml screw-top test tube filled with sterile liquid medium, tighten it and seal it with a parafilm, place it in a constant temperature incubator at 30°C and cultivate it for 5 days, until the culture medium is cloudy and gray-brown precipitates can be seen at the bottom, The activated bacterial liquid is obtained.

[0026] (2) Centrifuge 10 mL of the activated bacterial liquid obtained in step (1), remove the...

Embodiment 2

[0029] Embodiment 2: comparative test of static culture in incubator and stirring culture in fermenter

[0030] Get the 3rd enlarged culture bacterium liquid that embodiment 1 makes and carry out following experiment:

[0031] Static culture in the incubator: Inoculate the third expanded culture solution into 1L of sterilized ATCC1653MSGM medium (pH6.75) according to the inoculum amount of 5%, the iron source addition amount is 0.02μmol / L, and the temperature is static at 30°C. Place and cultivate for 120h, measure the growth curve and Cmag value at the same time, the results are as follows: figure 1 As shown, after 48 hours of static culture, the OD value of the bacterial cell density was 0.879. The bacteria were collected by centrifugation, ultrasonically crushed to obtain magnetosomes, and weighed after purification. The obtained magnetosomes had a wet weight of 37.1 mg, a dry weight of 0.3 mg, and a yield of 0.3 mg / L.

[0032] Stirring culture in fermenter: according to ...

Embodiment 3

[0033] Embodiment 3: the influence of different iron source additions in fermentor culture AMB-1 thalline

[0034] This example is basically the same as the fermenter stirring culture experiment in Example 2, except that the addition of iron source is divided into three levels: 0.02, 0.2 and 2 μmol / L, and three batches of fermentation culture are carried out. During the cultivation process, samples were taken to measure the OD at the beginning and end and the increment of OD value. The results are shown in Table 1. The wet weights of magnetosomes obtained in the two batches of iron source additions of 0.02 and 0.2 μmol / L were 190.1 and 235.0 mg, the dry weights were 22.7 and 63.3 mg, and the yields were 7.6 and 21.1 mg / L, respectively. The amount of iron source added is 2 μmol / L, and the amount of magnetosomes obtained in batch experiments is extremely low, so it cannot be weighed.

[0035] In order to prove whether AMB-1 synthesizes magnetosomes, observe the arrangement of m...

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Abstract

The invention discloses a culture method of magnetotactic bacteria AMB-1. The culture method comprises the following steps that 1, activating bacteria; 2, obtaining first enlarged culture bacteria; 3, obtaining second enlarged culture bacteria; 4, obtaining third enlarged culture bacteria; 5, taking a proper amount of the third enlarged culture bacteria liquid obtained in the step 4 to be inoculated into a sterile fluid nutrient medium 1653 MSMG, and conducting fermenter culture; 6, conducting centrifugation to collect the bacteria in the materials obtained in the step 5, and conducting ultrasonication washing, purification and freeze drying to obtain magnetosomes. According to the method, the aim that the AMB-1 bacteria density and the magnetosome yield are greatly improved though fermenter culture can be achieved under the conditions that only the addition amount of an iron source is changed and pH is controlled to be constant.

Description

technical field [0001] The invention relates to the technical field of microorganism culture, and in particular relates to a method for cultivating magnetotactic bacteria AMB-1. Background technique [0002] Magnetotactic bacteria are bacteria that can use geomagnetism or an external magnetic field to "navigate" and orient organisms. Magnetosomes are biofilm-coated nanoscale magnetic crystal particles synthesized in the cells and arranged in chains in the cells. The magnetotactic bacterium Magnetospirillum magneticum AMB-1 is a spiral magnetotactic bacterium isolated from the sediments of natural freshwater springs in Tokyo by the Japanese scholar Mataunaga. It is one of the few strains that can be cultivated under laboratory conditions and can produce Fe. 3 o 4 magnetosome particles. The magnetosomes synthesized by magnetotactic bacteria are nano-scale single magnetic domain crystals, non-cytotoxic, and have the characteristics of relatively large surface area and volume...

Claims

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Application Information

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IPC IPC(8): C12N1/20
CPCC12N1/20
Inventor 刘源岗洪雅真王士斌于汝祺刘华青
Owner HUAQIAO UNIVERSITY
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