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.
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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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