Marine fungi aspergillus terreus C23-3, fermentation liquor active extract as well as preparation method and application thereof
A technology for marine fungi and Aspergillus terreus, applied in the field of microorganisms, can solve the problems of high price, large toxic and side effects of drugs, and achieve the effects of anti-oxidation and inhibition of acetylcholinesterase activity, and good application prospects.
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Embodiment 1
[0060] Example 1 Isolation and Identification of Marine Fungus Aspergillus terreus C23-3
[0061] 1. Isolation method of strain
[0062] (1) Sample: peony coral in the South China Sea.
[0063] (2) Separation method
[0064] Wash the coral 3 times with sterile seawater, grind it to a muddy state with a sterilized mortar and pestle at room temperature and aseptic environment, take a little (about 1g) into 9 mL with a sterilized medicine spoon In sterile seawater, add glass beads to shake for 1 min, then let it stand for 20 min, and take the supernatant to make 10 -1 、10 -2 、10 -3 、10 -4 1-fold dilution, respectively draw 10 -1 、10 -2 、10 -3 、10 -4 Put 100 μL of the diluted sample dilution on the corresponding separation medium plate, spread evenly on the surface of the corresponding medium with a sterile glass coating rod, and culture in a constant temperature incubator at 28°C for 3 to 5 days. Then use a sterile razor blade to cut off the tip of the mycelium from the...
Embodiment 2
[0075] Example 2 Activity test of total extract from fermentation of marine fungus Aspergillus terreus C23-3
[0076] 1. Obtaining the total extract from fermentation of Aspergillus terreus C23-3
[0077] (1) Seed culture: Inoculate the strains from the slope onto a sterile seed solid medium plate, place the culture plate in a constant temperature (28°C) constant humidity (70%) incubator upside down for culture, and the culture time 4 to 5 days;
[0078] (2) Expansion culture: fill 1L of liquid medium into a 3L Erlenmeyer flask, seal it with a gas-permeable sealing film, and then sterilize in an autoclave at 121°C and 0.1 Mpa for 20 minutes; after cooling, inoculate the above-mentioned Step (1) After cultivation, the seed flat plate bacteria-carrying medium block is 30-40 cm 2 The proportion of inoculation, sealing, placed in a constant temperature of 28 ℃ indoor culture, natural light, culture time is 21 to 24 days, to obtain fermentation broth.
[0079] Wherein, the cultu...
Embodiment 3
[0092] Example 3 Extraction of Active Substances from Marine Fungus Aspergillus terreus C23-3 Fermentation Broth
[0093] 1. Medium
[0094] (1) Seed solid medium: add 20.0g sucrose, 20.0g sea salt, 5g peptone and 15.0g agar to every 500mL potato juice, pH is 7.4±0.2;
[0095] (2) The liquid medium is: add 20.0g sucrose, 20.0g sea salt and 5g peptone to every 500mL potato juice, the pH is 7.4±0.2;
[0096] Among them, potato juice: take fresh potatoes, peeled and cut into 1cm 3 For every 200g of potato pieces, add 500 mL of distilled water and boil for 20 minutes, filter through double-layered gauze cloth, and dilute to 500 mL with distilled water.
[0097] 2. Extraction method
[0098] (1) Seed culture: Inoculate the strains from the slope onto a sterile seed solid medium plate, place the culture plate in a constant temperature (28°C) constant humidity (70%) incubator upside down for culture, and the culture time 4 to 5 days;
[0099] (2) Expansion culture: fill 1L of li...
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