A kind of preparation method and use of secondary metabolites of endophytic fungus chaetomium sp of Eucalyptus margina
A technology of secondary metabolites and endophytic fungi, applied in the direction of microorganism-based methods, biochemical equipment and methods, botany equipment and methods, etc.
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experiment example 1
[0050] Experimental example 1 Fermentation of endophytic fungus Chaetomium sp.
[0051] First, pick a little endophytic fungus Chaetomium sp. from the 4°C cryotube Inoculate on the PDA solid medium plate for activation and culture for 5-7 days, subculture for 4-5 days, then inoculate the mycelia in a 500mL Erlenmeyer flask filled with 200mL PDB liquid medium, place at 25°C, 150rpm Shake culture for 6-7 days. The obtained liquid seeds were inserted into the sterilized rice culture medium, and then transferred to a constant temperature incubator at 28°C for fermentation for 60 days, and a total of 6Kg of rice culture medium was fermented.
experiment example 2
[0052] Preparation of Experimental Example 2 Secondary Metabolites
[0053] (1) Preparation of ethyl acetate layer crude extract
[0054] The endophytic fungus Chaetomium sp. solid fermentation product was taken out from the fermentation bottle and placed in the extraction barrel, the fungus block was smashed up with a shovel, poured into an appropriate amount of methanol for cold extraction, and the extraction was repeated 3 times, each time for 3 days. The filtrate was obtained by filtering with gauze, the fermentation broth was vacuum filtered, and concentrated to dryness under reduced pressure to obtain a crude extract. The crude extract was suspended in water, extracted with petroleum ether and ethyl acetate, and then concentrated under reduced pressure to obtain the crude extract of petroleum ether layer (15.15 g) and the crude extract of ethyl acetate layer (51.40 g), respectively.
[0055] (2) column chromatography under reduced pressure of ethyl acetate layer crude e...
experiment example 3
[0077] The antibacterial activity of experimental example 3 secondary metabolites
[0078] (1) Test bacteria
[0079] The selected test bacteria were: Staphylococcus aureus (S.aureus ATCC29213 and drug-resistant strain S.aureus N50) (G + ), Escherichia coli (G - ), Agrobacterium tumefaciens ATCC 11158 (G - ), Pseudomonas lachrymansATCC 11921 (G - ), Ralstonia solanacearum (G - ) and tomato scab (Xanthomonasvesicatoria ATCC 11633) (G - ).
[0080] (2) Solution preparation
[0081] Phosphate buffer solution (PBS) configuration: use NaH 2 PO 4 and Na 2 HPO 4 Prepare 0.2 mol / L PBS buffer solution with pH=7.2.
[0082] Preparation of thiazolium blue (MTT) solution: the above-mentioned PBS buffer solution was prepared into a MTT solution with a concentration of 5 mg / mL.
[0083] Configuration of sample solution: Accurately weigh 2.0 mg of each secondary metabolite separated above and dissolve in 0.3 mL of acetone, then add 0.7 mL of distilled water to make a mother solut...
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