Method for preparing epipodophyllotoxin diglucoside and special strain used thereby
A technology of epipodophyllotoxin and diglucoside, which is applied in the field of preparation of epipodophyllotoxin diglucoside, which can solve the problems of wild podophyllophyllous plants with complex growth environment, difficult to simulate, long growth cycle, etc., and achieve the protection of ecological diversity of plants The effect of high reliability, strong operability and short production cycle
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Embodiment 1
[0024] Embodiment 1, isolation and identification of bacterial strain
[0025] 1. Separation
[0026] The roots and rhizomes of the fresh plant Sinopodophyllum hexandrum (Royle) Ying were collected from Dacaotan (2700m above sea level) in Zhang County, Gansu Province. Take the root and rhizome of the fresh plant Taoerqi, roughen the surface, and wash it with tap water. In the ultra-clean bench, treat with 75% ethanol for 5 minutes, rinse with sterile water for 3-5 times, then treat with 2.5% sodium hypochlorite for 10 minutes, and rinse with sterile water for 3-5 times. Use sterilized tweezers and razor blades to peel off the outer skin of the material, then cut into small pieces of 0.5cm×0.5cm and plant them on PDA solid medium, and cultivate them at 28°C for 3-7 days. At the same time, the same sterilized materials were not peeled and cut, rolled on the PDA medium for one week, cultured under the same conditions, and observed as a control.
[0027] After a few days of cul...
Embodiment 2
[0039] Embodiment 2, preparation epipodophyllotoxin diglucoside
[0040] 1. Culture strains
[0041] Put 100ml of prepared potato dextrose liquid culture medium in a 250ml Erlenmeyer flask, sterilize, pick mycelia or spores and inoculate in the Erlenmeyer flask, culture at 28°C and 120r / min on a shaker for 7 days to obtain a fermentation broth.
[0042] Potato dextrose liquid culture medium is prepared according to the following method: Weigh 200g of potatoes (peeled, cut into about 2cm 2 small pieces), boil in water for 30 minutes, then filter with double gauze, take the filtrate and add 20g of glucose, then make up water to 1000mL, natural pH.
[0043] The experiment was repeated 3 times.
[0044] Second, the extraction of the product:
[0045] Use a suction filter to filter the fermentation broth twice, collect mycelia and fermentation broth respectively; extract the fermentation broth with an equal volume of chloroform, and use TLC to detect the chloroform extract. The...
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