A Bacillus coagulans that regulates allobaculum
A technology of Bacillus coagulans and dry bacteria, which is applied in the field of microorganisms to achieve the effects of strong spore-producing ability, broad application prospects and good tolerance.
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Embodiment 1
[0036] Example 1 Bacillus coagulans spore-producing ability
[0037] (1) Activation of bacteria
[0038] Bacillus coagulans WX12, LY11 or NJ23 was inoculated into MRS medium at an inoculation amount of 2 mL / 100 mL, and cultured at 37°C for 18 hours to obtain activated Bacillus coagulans strains, which were continuously activated for 2 generations. Colonies of Bacillus coagulans NJ23 are characterized by good growth on MRS medium, forming beige, opaque, round colonies (see figure 1 ). The characteristics of the bacteria are that the bacteria are rod-shaped, with blunt ends, arranged in single, pair or chain, the Gram staining reaction is positive, and the spores are terminal ( figure 2 ).
[0039] (2) Preparation of spores
[0040] Inoculate the above-mentioned activated strains into sporulation medium with an inoculum amount of 2mL / 100mL, culture in a shaker flask at 250r / min at 37°C for 48h, heat in a water bath at 85°C for 13min, and then serially dilute, MRS medium Po...
Embodiment 2
[0045] Example 2 Bacillus coagulans spore tolerance to simulated gastrointestinal fluid
[0046] Preparation of simulated gastric juice: Dissolve 0.3g of pepsin (1:15000, SIGMA) in 100mL of sterile normal saline (pH adjusted to 3.0), filter through a 0.22um sterile filter, and prepare immediately for use.
[0047] Prepare simulated intestinal juice: Dissolve 0.1g trypsin (1:250, Sangon) and 0.3g bile salt (BIOLEAF) in 100mL sterile normal saline (adjust pH to 8.0), filter with 0.22μm sterile filter membrane, and prepare now use.
[0048] The spores were prepared according to the method of Example 1, centrifuged at 6000g for 10 min, discarded the supernatant, washed 3 times with sterile normal saline, and resuspended in sterile normal saline until the concentration of the spores was 1×10 7 cfu / mL to obtain the initial spore fluid. Take 5mL of the initial spore liquid and centrifuge to discard the supernatant, resuspend with 5mL of simulated gastric juice, incubate at 37°C for...
Embodiment 3
[0052] Embodiment 3 Germination ability of bacillus coagulans spores in simulated gastrointestinal environment
[0053] The spores of Bacillus coagulans basically have no metabolic effect, and only maintain potential germination ability. After they germinate into vegetative cells, the metabolites produced by growth and reproduction play a probiotic role. Germination capacity in the environment.
[0054] The spore itself has a high degree of refraction. Once the spore germinates, the refraction will disappear immediately. The spectrophotometer detects the optical density change at the wavelength of 600nm, which reflects the germination rate of the average germination of the group of spores. The preparation of the simulated gastrointestinal fluid is the same as in Example 2, and the spores are prepared according to the method of Example 1, centrifuged at 6000 g for 10 min to discard the supernatant, washed 3 times with sterile normal saline, and resuspended in sterile normal sal...
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