A kind of marine fungi-bacteria symbiont and its metabolite and its application in the preparation of antibacterial drugs
A symbiotic, fungal technology, applied in the fields of chemicals for biological control, antibacterial drugs, microorganism-based methods, etc., can solve problems such as no medicine, residue, serious environment, etc. The effect of antifungal activity, reliable source and simple process
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Embodiment 1
[0031] Embodiment 1: Separation and purification of bacterial strains
[0032] Take an appropriate amount of seabed silt from Daya Bay in the northern part of the South my country Sea, put it in a clean petri dish, and air-dry it for later use (about a week). Put appropriate amount of air-dried sea mud and fresh sea mud into a 250mL Erlenmeyer flask filled with 30mL of artificial seawater (a small amount of glass beads can be added inside), shake at 200rpm on a shaker at 28°C for 30min, and then heat-shock in a water bath at 55°C. 5min standby. Use a pipette gun to draw 350 μL of the prepared sea mud sample, and evenly spread it on the plate of the prepared fucose-proline agar. The formula of the medium is: proline 1g, fucose-proline 5g, K 2 HPO 4 1g, MgSO 4 .7H 2 O 1g, (NH 4 ) 2 SO 4 1g, multivitamin (p-aminobenzoic acid, riboflavin, folic acid, inositol, pantothenic acid, Vc, VB1, VB6, VB12 50mg each, biotin 25mg), sea salt 30g, CaCO 3 2g, 20g of agar powder, add ...
Embodiment 2
[0033] Embodiment 2: the identification of bacterial strain
[0034] 1. Morphological identification
[0035] The obtained culture host fungus F190 (fungus / bacteria symbiont Spiromastix sp.SCSIO F190 / Alcaligenes faecalis SCSIO B001) bacteria cake was inoculated onto the poured ISP2, ISP4 and MEA plates, cultured in the dark at 28°C, and the growth morphology of the colonies was observed regularly every day . The sporulation structure and the morphology of the spores were observed with an optical microscope and a scanning electron microscope, and the size of the spores was measured. Its ecological characteristics are as figure 2 As shown, the colony grows faster on the ISP2 medium, and the diameter of the colony is 1.0-3.5mm in 3 days under the dark condition of 28°C. The colony is white at the beginning, and then gradually turns orange. Such as figure 2 A-D); The growth of the colony on the ISP4 medium is slow, and the diameter of the colony is smaller for 3 days at 28°C...
Embodiment 3
[0042] Embodiment 3: Preparation of the fermentation broth of fungi / bacteria symbiont Spiromastix sp.SCSIO F190 / Alcaligenes faecalisSCSIO B001
[0043] Pick an appropriate amount of fungal / bacterial symbiont Spiromastix sp.SCSIOF190 / Alcaligenes faecalis SCSIO B001 from the prepared ISP2 medium plate and add it to 50mL of PDB medium, culture at 28°C and 200r / min on a shaker for 2-3 days Obtain seed liquid. Then carry out expanded fermentation, add all 50mL seed liquid into a 1L Erlenmeyer flask containing 200mL PDB medium, culture on a shaker at 28°C for 7 days at 200r / min to collect the fermentation liquid, centrifuge the fermentation liquid at 3900r / min to get the supernatant , that is, the strain fermentation broth is used for subsequent use.
[0044] Remarks: The PDB medium used above is the potato dextrose water medium purchased from Huankai Biotechnology Co., Ltd.
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