Method for improving yield of antibacterial lipopeptide bacillomycin D through fatty acid metabolism regulation
A fatty acid metabolism and fatty acid technology, applied in the field of microbial fermentation, can solve problems such as affecting the synthesis of bacillomycin D
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Embodiment 1
[0039] (1) Strain activation: streak the Bacillus subtilis fmbJ stored at -20°C on LA medium, place it in a 37°C constant temperature incubator for 16 hours, and pick single colonies in LB medium at 37°C, Activated overnight at 180 rpm.
[0040] (2) Seed culture: insert the activated bacterial liquid into the seed medium, and cultivate to OD at 37°C and 180rpm. 600 to 0.8-1.0.
[0041] (3) Fermentation culture: First, sodium propionate was prepared into a concentrated stock solution of 2.5 mol / L, and the sodium propionate solution was filtered and sterilized with a 0.22 μm organic filter membrane, and the final concentration was 0, 0.25 mmol / L, 0.5 mmol / L, 1mmol / L, 2.5mmol / L, and 5mmol / L were added to the sterilized fermentation medium. Then the seed liquid was inoculated into the fermentation medium of each group, and it was fermented at 33°C and 180rpm for 120h.
[0042] (4) Further screening of the added concentration: the sodium propionate solution is filtered and ster...
Embodiment 2
[0055] (1) Strain activation: streak the Bacillus subtilis fmbJ stored at -20°C on LA medium, place it in a 37°C constant temperature incubator for 16 hours, and pick single colonies in LB medium at 37°C, Activated overnight at 180 rpm.
[0056] (2) Seed culture: insert the activated bacterial liquid into the seed medium, and cultivate to OD at 37°C and 180rpm. 600 to 0.8-1.0.
[0057] (3) Fermentation culture: filter and sterilize propionic acid with a 0.22 μm organic filter membrane, add sterilized The pH of the fermentation medium was adjusted to about 7.0 with sterilized 4M NaOH. Then the seed liquid was inoculated into the fermentation medium of each group, and it was fermented at 33°C and 180rpm for 120h.
[0058] (4) Product detection: The fermentation broth was transferred to a centrifuge tube, centrifuged at 8000 rpm for 20 min, and the supernatant was collected. The pH of the supernatant was adjusted to 2.0 with 6M HCl and left at 4°C overnight. Centrifuge at 85...
Embodiment 3
[0061] (1) Strain activation: streak the Bacillus subtilis fmbJ stored at -20°C on LA medium, place it in a 37°C constant temperature incubator for 16 hours, and pick single colonies in LB medium at 37°C, Activated overnight at 180 rpm.
[0062] (2) Seed culture: insert the activated bacterial liquid into the seed medium, and cultivate to OD at 37°C and 180rpm. 600 to 0.8-1.0.
[0063] (3) Fermentation culture: filter and sterilize butyric acid with a 0.22 μm organic filter membrane, add sterilized The pH of the fermentation medium was adjusted to about 7.0 with sterilized 4M NaOH. Then the seed liquid was inoculated into the fermentation medium of each group, and it was fermented at 33°C and 180rpm for 120h.
[0064](4) Product detection: The fermentation broth was transferred to a centrifuge tube, centrifuged at 8000 rpm for 20 min, and the supernatant was collected. The pH of the supernatant was adjusted to 2.0 with 6M HCl and left at 4°C overnight. Centrifuge at 8500r...
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