Bacillus subtilis and application
A Bacillus subtilis, strain technology, applied in bacteria, medical preparations containing active ingredients, antidote, etc., to achieve the effect of good colonization and great application potential
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Embodiment 1
[0062] This example provides the screening, isolation and detection of the strain with the preservation number CCTCC No: M 20211386. The specific operations are as follows:
[0063] (1) 85 strains were isolated from selenium-rich soil in Enshi, Hubei. Strain isolation: uniformly disperse the collected soil sample particles on LB solid medium, and culture at 37°C. Single colonies were transferred the next day. Preliminary screening: The single colonies isolated from above were inoculated into LB plates containing 9, 17, 26, 35, 43, 52 mg / mL sodium selenite respectively. According to whether there are reddish to dark red colonies on the plate, select the strains that can produce dark red colonies for re-screening.
[0064] Re-screening: The seed liquids of different strains after primary screening were inoculated in LB liquid medium containing 9, 17, 26, 35, 43, 52 mg / mL sodium selenite respectively. Shake culture at a constant temperature at 37°C, observe whether the color of...
Embodiment 2
[0073] In this embodiment, the Bacillus subtilis screened out in Example 1 is tested for performance, and the specific operations are as follows:
[0074] (1) Transfer about 1ml of overnight-grown bacterial solution to 9ml of fresh LB medium adjusted to pH 2.5 and 3.5 with hydrochloric acid (LB liquid medium: peptone 10g / L, yeast extract 5g / L, NaCl 10g / L) and incubated at 37°C for 4h. Similarly, for bile salt tolerance, the T5 strain was transferred to LB medium containing 0.3w / v% and 0.6w / v% bovine bile salt and incubated at 37°C for 4h. At 0 and 4 hours of incubation, the number of viable cells was determined by serially diluting the cultures and then plating them on LA plates. Viability was determined by the number of viable cells present on the agar and was calculated as colony forming units (CFU) per milliliter of sample. The results showed that the survival rate of T5 could reach 70.63% in the environment of pH 3.5.
[0075] (2) Take the T5 bacterial suspension whose ...
Embodiment 3
[0080] Embodiment 3 utilizes the T5 bacterial strain that embodiment 1 separates to prepare nano-selenium, and concrete operation is as follows:
[0081]Inoculate the activated T5 strain into the LB medium of sodium selenite, the inoculum amount is 1v / v%, culture at 37°C with constant temperature shaking, collect the red bacterial liquid, centrifuge at 9000rpm for 20min, and collect the precipitate. Grind the precipitate with liquid nitrogen, ultrasonically pass the red suspension through 20, 10, 5, 3, 1.2, and 0.8 μm filter membranes respectively, fully extract the filtered red suspension with n-hexane, collect the lower layer, centrifuge at 10,000 rpm for 10 min, and collect The precipitate is the product, freeze-dried and weighed. The concentration and incubation time of the sodium selenite in Example 3 are shown in Table 2. The nano-selenium product in Example 3 was added to the pre-cooled electron microscope fixative solution and fixed overnight at 4°C. Fixture is carrie...
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