A bifidobacterium strain that can be used for selenium-enriched culture and its active protein and application
A bifidobacterium and active protein technology, applied in the field of microorganisms, can solve the problems of affecting the intestinal colonization of bifidobacteria, reducing proton power, and destroying the osmotic pressure balance of bifidobacteria cells, so as to achieve good intestinal colonization ability, Effects of hepatoprotection, good gut protection and hepatoprotective function
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Embodiment 1
[0040]The acquisition of embodiment 1 bifidobacterium DD98 and strain isolation, screening
[0041] 1. Sample collection and expansion culture
[0042] Take 1g of the fresh mid-section feces of the sampled subjects, add 10ml of normal saline, mix well, let stand at room temperature for 5min, and collect the upper suspension. Take 1ml of the supernatant suspension, inoculate it into 50ml of RCM medium, and incubate anaerobically at 37°C for 24h to obtain a culture solution.
[0043] 2. Strain Isolation
[0044] The culture solution was taken and diluted 10 times with normal saline -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 times, to obtain a bacterial suspension, pipette 0.1ml of the bacterial suspension and spread it on the BBL medium, and culture it anaerobically at 37°C for 24 hours to obtain a single colony.
[0045] 3. Strain purification
[0046] Pick a single colony, streak and inoculate the colony culture on the BBL medium, and inoculate it anaerobically at 37°C for 24 ...
Embodiment 2
[0051] Example 2 Selenium-enriched domestication investigation of bifidobacterium DD98
[0052] Prepare a sodium selenite solution, filter it at 0.22um and add it to the culture medium, then distribute the culture medium to 96 deep-well plates and plates. Preliminary design of selenium-enriched acclimation gradient: 5, 10, 15, 20, 25, 50, 100, 200, 400, 600, 800 μg / ml.
[0053] Divide glycerol tubes of dominant strains of bifidobacteria into 96 deep-well plates containing 5 μg / ml sodium selenite medium, culture anaerobically for 24 hours, take 200 μl from each well of 96 deep-well plates to 96 microplate plates for OD600 For detection, the 3 wells with the highest OD value were diluted to 10 5 ~10 6 Double coated onto 5μg / ml sodium selenite plate. Put into anaerobic incubator and cultivate 18 hours after plate coating, after treating that single bacterium colony grows, pick the single bacterium colony (illustration that this bacterium colony grows well on the sodium selenit...
Embodiment 3
[0054] The bacteriological characteristic of embodiment 3 bifidobacteria DD98
[0055] 1. Colony Morphological Characteristics
[0056] Observing the isolated and purified DD98 with a scanning electron microscope, the bacteria are long rod-shaped or branched rod-shaped at the end, and the bacteria are generally about 10um in length, such as figure 1 shown.
[0057] The morphological characteristics of the colony: neat edges, soft texture, smooth surface, milky white protrusions, such as figure 2 shown.
[0058] Gram staining optical microscope observation: the staining result is purple, such as image 3 Shown, indicating Gram-positive bacilli.
[0059] 2. Biochemical characteristics
[0060] Using Bergey's Handbook as a reference, the characteristics of sugar fermentation and acid production of DD98 were investigated, and the results are as follows:
[0061]
[0062]
[0063] The results showed that: Bifidobacterium DD98 can utilize L-arabinose, D-galactose, D-fru...
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