Preparation method of active matter capable of inhibiting legionella in bacillus HZ16
A technology of Bacillus and HZ16, which is used in the preparation of bioactive components by fermentation, the application of new drug development, the active field of Bacillus strains, and can solve the problems of difficult treatment and killing of other diseases.
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Embodiment 1
[0028] The preparation method of the active substance inhibiting Legionella in Bacillus HZ16 is prepared according to the following steps:
[0029] Step 1, inoculate the purified Bacillus HZ16 on the LB solid medium with pH 5 under aseptic conditions, and cultivate at 28 degrees Celsius for 1 day to obtain white solid colonies;
[0030] Step 2, inoculate the solid bacterial strain into the 300 ml conical flask containing the LB substratum of 5 with 150 ml pH;
[0031] Step 3, placing the inoculated shake flask in a biological incubator, and culturing it under the conditions of 28 degrees Celsius and 180 r / min for 1 day-based fermentation to obtain a fermentation broth;
[0032] Step 4, utilize freeze-drying technology to remove moisture, compare the solid dry weight obtained;
[0033] Step 5, extracting crude extract with organic solvent ethyl acetate, and concentrating for subsequent use;
[0034] Step 6. It is verified that the biologically active peptide obtained in Examp...
Embodiment 2
[0037] Step 1. Inoculate the purified Bacillus HZ16 on the LB medium with pH 8 under aseptic conditions, and cultivate at 28 degrees Celsius for 1 day to obtain white solid colonies;
[0038] Step 2, inoculate solid mycelium to containing 150 ml of pH 8 M 2 50% SW medium in a 300 ml Erlenmeyer flask;
[0039] Step 3, placing the inoculated shake flask in a biological incubator, and culturing it for 4 days under the conditions of 28 degrees Celsius and 200 r / min to obtain a fermentation broth;
[0040] Step 4, utilize freeze-drying technology to remove moisture, compare the solid dry weight obtained;
[0041]Step 5, extracting crude extract with organic solvent ethyl acetate, and concentrating for subsequent use;
[0042] Step 6: Dissolving the crude extract in DMSO to prepare a solution of 50 μg / mL, and performing the same activity test for inhibiting Legionella as in Step 6 in Example 1, the 48-hour inhibition circle size was 22 mm.
Embodiment 3
[0044] Step 1, inoculate the purified Bacillus HZ16 on the high's medium with pH 8 under aseptic conditions, and cultivate at 28 degrees Celsius for 4 days to obtain white solid colonies;
[0045] Step 2, inoculate solid mycelium into 300 milliliters of conical flasks containing 150 milliliters of pH 8 Gao's culture medium;
[0046] Step 3, placing the inoculated shake flask in a biological incubator, and culturing it for 4 days under the conditions of 28 degrees Celsius and 200 r / min to obtain a fermentation broth;
[0047] Step 4, utilize freeze-drying technology to remove moisture, compare the solid dry weight obtained;
[0048] Step 5, extracting crude extract with organic solvent ethyl acetate, and concentrating for subsequent use;
[0049] Step 6: Dissolving the crude extract in DMSO to prepare a solution of 50 μg / mL, and performing the same activity test of inhibiting Legionella as in Step 6 in Example 1, the 48-hour inhibition circle size is 18 mm.
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