Endophytic fungus strain of Huperz serrata and its application
A technology of endophytic fungi and Huperzia serrata, applied in the field of microbiology and chemistry, can solve the problem of unclear pathogenesis of Alzheimer's disease
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Embodiment 1
[0034] Example 1 Separation and purification of the endophytic fungal strain Eurotiaceae Penicillium sp.L10Q37 of Huperz serrata according to the present invention
[0035] Take fresh Huperzia serrata (melaleuca) root section, rinse it, immerse it in 70% alcohol for disinfection for 4 minutes, rinse it with sterile water for 5 times, soak it in 1% sodium hypochlorite solution for 5 minutes, and then rinse it with sterile water for 5 times , then plant the root section on the PDA medium, put it in a 28°C incubator and cultivate it for 2 weeks, take the explants without fungal contamination and sterilize them with 1% sodium hypochlorite solution for 15 minutes, cut them and inoculate them in the PDA medium for cultivation. Use an inoculation needle to pick and inoculate the hyphae growing from the edge of the sample on a PDA medium plate, culture at a constant temperature of 28°C, and after new colonies grow, pick the tip mycelium and transfer it to a PDA medium plate to repeat p...
Embodiment 2
[0036] Embodiment 2 Shaking bed cultivation method of the endophytic fungus strain Eurotiaceae Penicillium sp.L10Q37 of Huperz serrata of the present invention
[0037] PDA liquid culture medium: the method is to wash and peel the potatoes first, then weigh 200g of potatoes and cut them into small pieces, add water and boil them (boil for 20-30 minutes, it can be pierced by a glass rod), filter with eight layers of gauze, Add 20g of glucose to the filtrate, stir evenly, and then replenish the water to 1000ml after cooling slightly, and the pH is natural. Divide into test tubes or Erlenmeyer flasks, stopper and bandage, sterilize at 121°C for about 20 minutes, take out and shake well, cool and store for later use.
[0038] PDA solid medium: the method is to wash and peel the potatoes first, then weigh 200g of potatoes and cut them into small pieces, add water and boil them (boil for 20-30 minutes, it can be pierced by a glass rod), filter with eight layers of gauze, Heat, add ...
Embodiment 3
[0041] Embodiment 3 Fermentation method of the endophytic fungus Eurotiaceae Penicillium sp.L10Q37 of Huperz serrata according to the present invention
[0042] Preparation of seed solution: Aseptic operation, pick the activated slant strains in Example 2, inoculate them in PDA liquid medium, and culture them on a constant temperature shaker at 28° C. at 150 rpm for 48 hours. The fermented liquid was filtered with 4 layers of sterilized gauze to obtain a uniform first-grade seed liquid. The primary seeds were inserted into the liquid PDA medium with an inoculum amount of 10%, and cultured on a constant temperature shaker at 150 rpm at 28° C. for 48 hours to obtain secondary seed liquid.
[0043] Fermentation and cultivation process: Add 20L liquid PDA medium into a 30L ventilated stirring fermenter, sterilize at 121°C for 20 minutes and cool to 28°C, add 1% of the secondary seed liquid, and ferment to obtain a fermentation product. The fermentation parameters are: ventilation...
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