Application of mycelium extract of strain culture of marine fungus umbelopsis sp. in preparation of antibacterial drugs
A technology of marine fungi and mycelium, which is applied in antibacterial drugs, fungi, medical raw materials derived from fungi, etc., can solve the problems of large toxic side effects, strong drug resistance, and lack of antifungal drugs.
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Embodiment 1
[0036] Example 1: Isolation and identification of the marine fungus Umbelopsis sp. QZ042
[0037] 1. Isolation of Fungi
[0038] Take a sample of marine rot wood in a sterile conical flask, weigh 5g, mash it with a sterile glass rod, add 50ml of sterile saline, add glass beads and shake on a shaker for 20min, after standing for stratification, take the upper layer to make 10 times Gradient dilution, take the original solution, 10 times and 100 times dilutions Tubu PDA solid medium, culture at a constant temperature of 26 ~ 30 ℃, after the colonies grow, pick the colonies and streak to purify.
[0039] 2. Fungal culture and morphological observation
[0040] Observation of culture characteristics: the strains were inoculated on PDA medium, and cultured at 26°C to obtain colonies. Record the color of the primary hyphae, the color change of the colony, the surface characteristics, and the presence or absence of pigmentation.
[0041] Film production and microscopic examination...
Embodiment 2
[0056] Embodiment 2: Extraction of active substance of marine fungus Umbelopsissp. QZ042
[0057] 1. Seed culture
[0058] Medium: 180-220g of potatoes, 18-22g of glucose, 20g of agar, 1000mL of 50% artificial seawater. Fungal hyphae were picked and inoculated on the culture medium for slant culture, and cultured at 28°C for 5-7 days. Wherein, the formula of artificial seawater (salinity 3.34%) is as follows: every liter of water contains: sodium chloride NaCl26.726g, magnesium chloride MgCl 2 2.26g, magnesium sulfate MgSO 4 3.248g, calcium chloride CaCl 2 1.153g, sodium bicarbonate NaHCO 3 0.198g, potassium chloride KCI0.721g, sodium bromide NaBr0.058g, boric acid H 3 BO 3 0.058g, sodium silicate Na 2 SiO 3 0.0024g, sodium silicate Na 2 Si 4 o 9 0.0015g, phosphoric acid H 3 PO 4 0.002g, aluminum hexachloride Al 2 Cl 6 0.013g, ammonia NH 3 0.002g, lithium nitrate LiNO 3 0.0013g.
[0059] 2. Fermentation culture
[0060] Fermentation medium: 180-220g of potat...
Embodiment 3
[0064] Example 3: Antibacterial activity of mycelium extracts from cultures of the marine fungus Umbelopsis sp. QZ042
[0065] 1. Preparation of indicator bacteria
[0066] Inoculate E.Coli, B.cereus, Arthrobacternicotianae, Klebsiellapeneumoniae, and Vibrioparahaemolyticus in beef extract-peptone liquid medium respectively, and culture them overnight on a shaker at 37°C at a speed of 150r / min. The indicator fungi Eurotiumrubrum, Alternariasp., Lasiodiplodiapseudotheobromae, Diaporthephaseolorum, and Pestalotiopsismicrospora were inoculated on PDA plates and cultured to 1 / 2 of the petri dish. These bacteria and fungi are preserved by our laboratory. Inoculate 2 plates for each indicator strain.
[0067] 2. Determination of Antibacterial Activity
[0068] Weigh the mycelium extract prepared in Example 2, and dilute the solution to 400 μg / ml, 200 μg / ml, 100 μg / ml, 50 μg / ml, 25 μg / ml, 12.5 μg / ml, 6.25 μg by adding sterile water / ml, and its antibacterial activity was determin...
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