Cephalosporium acremonium corda rhzomorph, production method and uses thereof
A technology of spores and seeds, applied in the field of acremonium, to achieve the effect of guaranteed source, low cost and simple operation process
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Embodiment 1
[0026] Example 1: Separation and purification of Cephalosporium acremonium IFB-E007
[0027] Take fresh and healthy Trachelospermum root, put it into a fresh-keeping bag, seal and mark, and quickly take the sample to the laboratory (20°C), and isolate the endophytic bacteria within 5 hours. Wash the Trachelospermum root with clean water, cut into small pieces of about 1cm, and soak in 75% alcohol for 1min, 1% sodium hypochlorite solution (containing free chlorine>2.5%) for 10min, 75% alcohol for 1min, and then use the double antibody (200IU / mL) Penicillin and 150IU / ml streptomycin) WA medium (20g agar, distilled water 1000mL) plate culture. After the colony grows from the incision, it is transferred to PCA medium (20g potato, 20g carrot, 20g agar, distilled water 1000mL) for the isolation of endophytic fungi. The colony morphology observation, spore morphology and 18S rDNA molecular identification Cephalosporium acremonium, Cephalosporiumacremonium IFB-E007, preservation unit: Gen...
Embodiment 2
[0028] Example 2: Solid fermentation of Cephalosporium acremonium IFB-E007
[0029] Activate the plant endophyte Cephalosporium acremonium (Cephalosporium acremonium IFB-E007), inoculate the fresh mycelium mass into a 1000 mL conical flask, each flask contains 400 mL of PDA medium, cultured at 140 rpm, 28 ℃ for 3 days as Seed liquid: inoculate 20 mL of seed liquid in a jar containing solid medium, and ferment for 30 days at 28°C. The composition of the solid medium is 7.5g of millet, 7.5g of bran, and 0.5 of yeast extract in each bottle. g, FeSO 4 ·7H 2 O 0.01g, sodium tartrate 0.1g, sodium glutamate 0.1g, corn oil 0.1mL, distilled water 15mL.
Embodiment 3
[0030] Example 3: Extraction and Separation of Cephalosporin
[0031] The fermented product obtained in Example 2 was extracted 3 times at room temperature with chloroform / methanol (volume ratio 1:1), with a material-to-liquid ratio of 2:5 (g / ml), filtered and concentrated in a low-temperature vacuum to obtain a black crude extract F; The crude extract F was dissolved in methanol with a material-to-liquid ratio of 7:10 (g / mL). After heating and refluxing for 1 hour, it was cooled to room temperature and placed in a refrigerator at -18°C overnight. Oil and salt substances were repeated three times to obtain extract F1; the extract F1 was subjected to silica gel column chromatography, followed by 10 times the column retention volume of chloroform, chloroform / methanol (volume ratio 100:1), chloroform / methanol ( Volume ratio 100:2), elution with chloroform / methanol (volume ratio 100:4), combine the eluents eluted with chloroform / methanol (volume ratio 100:4) to obtain extract F2; add ...
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