Method for detecting nutrition environment change in subculture process of vitamin C production strain
A technology for the production of bacterial strains and environmental changes, applied in the field of industrial microorganisms, can solve problems such as unclear mechanism of action, achieve the effect of increasing production and optimizing the fermentation process
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Embodiment 1
[0040] A method for detecting changes in the nutritional environment during the subculture of vitamin C production strains, comprising the steps of:
[0041] (1) Subculture of mixed bacteria:
[0042] ① Solid culture:
[0043] Inoculate 500 μL of Gluconobacter oxydans stored in 15% aqueous glycerol and 500 μL of Bacillus megaterium stored in 15% aqueous glycerol in liquid nitrogen, respectively. Incubate on solid medium at 28°C for 24 hours;
[0044] ②Seed cultivation:
[0045] Transfer the Bacillus megaterium and Gluconobacter oxidans cultivated in step (1) into the seed culture medium respectively, and cultivate them on a shaker at 28°C and 200r / min for 24 hours to obtain the Bacillus megaterium seed liquid and Gluconobacter oxidans seed liquid ;
[0046] Inoculate the new seed medium with Bacillus megaterium and Gluconobacter oxydans so that the density of Bacillus megaterium is 2×10 7 CFU / mL, so that the density of Gluconobacter oxidans is 2×10 8CFU / mL, at 28°C, 200r...
Embodiment 2
[0077] A method for detecting changes in the nutritional environment during the subculture of vitamin C production strains, comprising the steps of:
[0078] (1) Subculture of mixed bacteria:
[0079] ① Solid culture:
[0080] Take 10 μL of Gluconobacter oxydans (Gluconobacter oxydans) preserved in 20% aqueous glycerol solution and 10 μL of Bacillus megaterium (Bacillus megaterium) preserved in 20% aqueous glycerol solution and inoculate them respectively. Incubate on solid medium at 30°C for 36 hours;
[0081] ②Seed cultivation:
[0082] Transfer the Bacillus megaterium and Gluconobacter oxidans cultured in step (1) into the seed culture medium respectively, and cultivate them on a shaker at 30°C and 250r / min for 36 hours to obtain the Bacillus megaterium seed liquid and Gluconobacter oxidans seeds respectively liquid;
[0083] Inoculate a new seed medium with Bacillus megaterium and Gluconobacter oxydans so that the density of Bacillus megaterium is 2 x 10 8 CFU / mL, so ...
Embodiment 3
[0101] A method for detecting changes in the nutritional environment during the subculture of vitamin C production strains, comprising the steps of:
[0102] (1) Subculture of mixed bacteria:
[0103] ① Solid culture:
[0104] Take 200 μL of Gluconobacter oxydans (Gluconobacter oxydans) preserved in 30% aqueous glycerol solution and 200 μL of Bacillus megaterium (Bacillus megaterium) preserved in 30% aqueous glycerol solution stored in liquid nitrogen and inoculate them respectively Incubate on solid medium at 35°C for 48 hours;
[0105] ②Seed cultivation:
[0106] Transfer the Bacillus megaterium and Gluconobacter oxidans cultivated in step (1) ① into the seed culture medium respectively, and culture at 35°C and 280r / min shaking table for 48 hours to obtain the Bacillus megaterium seed liquid and Gluconobacter oxidans seed liquid ;
[0107] Inoculate a new seed medium with Bacillus megaterium and Gluconobacter oxydans so that the density of Bacillus megaterium is 2 x 10 ...
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