Culture medium for culturing akkermansia muciniphila and use method thereof

A culture medium and mucin technology, applied in the field of culture medium for Akkermansia muciniphila, can solve the problems that Akk bacteria cannot be cultivated and failed.

Pending Publication Date: 2019-05-28
GUANGZHOU KANGZE MEDICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But carry out the amplified culture of Akk with the substratum reported in this document, all fail to obtain success, can not cultivate and obtain Akk bacterium

Method used

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  • Culture medium for culturing akkermansia muciniphila and use method thereof
  • Culture medium for culturing akkermansia muciniphila and use method thereof
  • Culture medium for culturing akkermansia muciniphila and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Take a 250ml Erlenmeyer flask, add 200mL of pure water, then add 0.72g of glucose, 2.4g of soybean peptone, 0.6g of threonine, 0.88g of N-acetylglucosamine, 0.9g of sodium chloride and 0.4g of dihydrogen phosphate Sodium, after fully stirring and mixing, the medium is autoclaved, and after the medium is cooled, inoculate 100 μL of Akkermansia muciniphila bacteria liquid into the medium (Akkermans muciniphila in the bacterial liquid The bacteria content is 2.6×10 8 cfu / mL) and 200 μL cysteine, placed in an anaerobic box (85% CO 2 ,10%N2 and 5%H 2 ) (cultivation temperature is 37°C) after 3 days, take it out, centrifuge at 3000rpm for 10min, observe the amount of precipitated bacteria, resuspend the precipitate with 5mL of sterile PBS, and use NanoDrop to measure the OD of the bacterial solution 600 value, OD 600 The value is 0.0183.

[0033] After the bacterial liquid was inoculated in the mucin liquid medium, cultured for 2 to 3 days, taken out and centrifuged, the ...

Embodiment 2

[0037] Take a 250ml Erlenmeyer flask, add 200mL of pure water, then add 0.72g of glucose, 3.2g of soybean peptone, 0.8g of threonine, 1.056g of N-acetylglucosamine, 1g of sodium chloride and 0.5g of disodium hydrogen phosphate , after fully stirring and mixing, the culture medium was autoclaved, and after the culture medium was cooled, 100 μL of Akkermansia muciniphila bacteria liquid (Akkermans muciniphila in bacteria liquid) was inoculated into the culture medium. The bacteria content is 2.6×10 8 cfu / mL) and 200 μL cysteine, placed in an anaerobic box (85% CO 2 ,10%N 2 and 5%H 2 ) (cultivation temperature is 37°C) after 4 days, take it out, centrifuge at 3000rpm for 10min, observe the amount of precipitated bacteria, resuspend the precipitate with 5mL of sterile PBS, and use NanoDrop to measure the OD of the bacterial solution 600 value, OD 600 The value is 0.0343.

[0038] Whether the bacteria in the bacterial solution is Akkermansia muciniphila and the detection of th...

Embodiment 3

[0040] Take a 250ml Erlenmeyer flask, add 200mL of pure water, then add 0.72g of glucose, 4g of soybean peptone, 1g of threonine, 1.328g of N-acetylglucosamine, 1.1g of sodium chloride and 0.6g of disodium hydrogen phosphate, After fully stirring and mixing, the culture medium was autoclaved, and after the culture medium was cooled, 100 μL of Akkermansia muciniphila bacteria liquid (the amount of Akkermansia muciniphila in the bacteria liquid) was inoculated into the culture medium. The bacteria content is 2.6×10 8 cfu / mL) and 200 μL cysteine, placed in an anaerobic box (85% CO 2 ,10%N 2 and 5%H 2 ) (cultivation temperature is 37°C) after 4 days, take it out, centrifuge at 3000rpm for 10min, observe the amount of precipitated bacteria, resuspend the precipitate with 5mL of sterile PBS, and use NanoDrop to measure the OD of the bacterial solution 600 value, OD 600 The value is 0.0423.

[0041] Whether the bacteria in the bacterial solution is Akkermansia muciniphila and th...

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Abstract

The invention provides a culture medium for culturing akkermansia muciniphila and a use method thereof, and belongs to the technical field of microorganism culture mediums. Components of the culture medium include soy peptone, threonine, glucose, N-acetylglucosamine, sodium chloride and disodium hydrogen phosphate. By adopting the culture medium, the akkermansia muciniphila quantity required by functional studying of the akkermansia muciniphila can be reached through culture, the components of the culture medium have safe sources, and potential biological hazards do not exist.

Description

technical field [0001] The invention belongs to the technical field of microbial culture medium, and in particular relates to a culture medium for cultivating Akkermansia muciniphila and a use method thereof. Background technique [0002] Obesity and its associated metabolic diseases such as diabetes and cardiovascular disease have become a major threat to public health. The situation in my country is particularly severe. There are more than 100 million diabetic patients, and nearly 500 million people are pre-diabetic. Therefore, it is particularly important to study measures to prevent and treat such diseases. [0003] Gut flora is closely related to human health. Obesity and insulin resistance are two important early features of metabolic diseases, and gut flora plays an important role in the occurrence and progression of metabolic diseases. Altered gut microbiota, inflammation, and disrupted gut barrier function are important features of obesity and type 2 diabetes. Ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 彭永正邓露露易江丰
Owner GUANGZHOU KANGZE MEDICAL TECH CO LTD
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