Fengyuansu, composition including Fengyuansu, fusion gene encoding Fengyuansu, recombinant plasmid, recombinant bacterium and application
A technology of Fengyuansu and composition, which is applied in Fengyuansu, recombinant bacteria and its application fields, and can solve problems such as poor control effect
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Embodiment 1
[0059] Example 1 Extraction of lipopeptide compounds produced by Bacillus velesi HN-Q-8
[0060] A single colony was cultured at 37°C for 24h by the cross streaking method, and the newly activated HN-Q-8 strain was inoculated into 100mL LB liquid medium for 24h at 37°C and 200r / min to obtain seed liquid; the seed liquid was inoculated to In 100mL LB liquid medium, the inoculum amount is 10%, and continue to cultivate at 37°C, 200r / min for 72h: centrifuge the fermentation broth at 4°C, 8000rpm for 20min, collect the supernatant, and slowly add 6mol / min to the supernatant. The pH was adjusted to 2.0 with 1 L of HCl, and the precipitate was left to stand at 4°C overnight; then, the precipitate was collected by centrifugation at 8000 rpm at 4°C for 20 min, and the precipitate was dried in a fume hood and 30 mL of methanol was added to fully dissolve the precipitate. The methanol extract was filtered through a bacterial filter to obtain lipopeptide compounds.
Embodiment 2
[0061] Example 2 Bacteriostatic activity of lipopeptide compounds produced by Bacillus velesi HN-Q-8
[0062] The ZB4 strain of Potato nevus was used as the model bacteria, and the concentrations of lipopeptides were formulated into 10 mg / mL, 5 mg / mL, 1 mg / mL, 0.5 mg / mL, and 0.1 mg / mL. The model pathogenic bacteria cake (5 mm) was inserted in the center of the plate, and holes were drilled on both sides 2.5 cm away from the bacterial cake. Each concentration was a treatment, and each treatment was repeated 3 times, and an untreated blank plate was used as a control. When the blank control group is about to fill the petri dish, observe and take pictures.
[0063] The result is as figure 1 shown, figure 1 A is the bacteriostatic activity of lipopeptides at a concentration of 10 mg / mL; B is the bacteriostatic activity of 5 mg / mL lipopeptides; C is the bacteriostatic activity of 1 mg / mL lipopeptides; D is 0.5 mg / mL lipopeptides bacteriostatic activity of lipopeptide compounds;...
Embodiment 3
[0064] Example 3 Determination of antibacterial spectrum of lipopeptide compounds produced by Bacillus velesi HN-Q-8
[0065] 1. Determination of the inhibition of lipopeptide compounds on pathogens: the concentration of 10 mg / mL was used for the determination of the antibacterial spectrum. Fungal bacteria (5 mm) were inserted into the center of the plate, holes were drilled on both sides 2.5 cm away from the bacterial cake, and 50 μL of methanol was added to one side, and 50 μL of lipopeptide compounds were added to the other side. Each bacteria was treated as one treatment, and each treatment was repeated three times, and an untreated blank plate was used as a control. When the blank control group covered the petri dish, it was observed and photographed.
[0066] 2. Determination of the inhibition of lipopeptides against bacteria and actinomycetes: the dilution factor is 1×10 -5 of potato blackleg and the dilution factor is 1×10 -2 Take 100 μL of bacterial liquid from eac...
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