Method for preparing epipodophyllotoxin diglucoside and special bacterial strain thereof

A technology for epipodophyllotoxin and diglucoside, which is applied in the field of preparing epipodophyllotoxin diglucoside, can solve the problems of complex growth environment, long growth cycle, and difficulty in simulation of wild podophyllotoxins, and achieves protection of plant ecological diversity. The effect of stability, short production cycle and strong operability

Inactive Publication Date: 2012-02-22
BEIJING NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The growth environment of wild podophyllum plants is complex, difficult to simulate, and the growth cycle is long, so it is difficult to carry out large-scale artificial cultivation, and tissue culture technology is not suitable for podophyllum plants; the chemical structure of podophyllotoxins is complex, and chemical synthesis Although it can be carried out, the cost is too high; genetic engineering means to construct engineered bacteria cannot be carried out due to the large number of enzymes involved in the formation of such substances.

Method used

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  • Method for preparing epipodophyllotoxin diglucoside and special bacterial strain thereof
  • Method for preparing epipodophyllotoxin diglucoside and special bacterial strain thereof
  • Method for preparing epipodophyllotoxin diglucoside and special bacterial strain thereof

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Experimental program
Comparison scheme
Effect test

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. Take...

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Abstract

The invention discloses a method for preparing epipodophyllotoxin diglucoside and a special bacterial strain thereof. The strain is Aspergillus parasiticus TEQB, and its preservation number is CGMCC No.3116. The bacterial strain of the present invention can be used to prepare epipodophyllotoxin diglucoside. The method for preparing epipodophyllotoxin diglucoside of the present invention has the advantages of simple operation, low cost, easy operation and short production cycle. The method of the present invention provides a new way for preparing epipodophyllotoxin diglucoside, avoids the limitations of other various preparation methods in the prior art, has strong operability, is suitable for industrialized large-scale production, and simultaneously It is of great significance to protect the biodiversity of plants.

Description

technical field [0001] The invention relates to a method for preparing epipodophyllotoxin diglucoside and a special bacterial strain thereof. Background technique [0002] Podophyllotoxin (podophyllotoxin) mainly exists in the Berberidaceae Podophyllum and its related plants. It is a class of natural compounds with a wide range of biological activities, including estrogen-like, anti-estrogen-like, anti-cancer, anti-viral , anti-fungal, sedative, inhibiting plant growth, anti-oxidation and other effects. In particular, it has strong anticancer activity, and its mechanism of action is to inhibit the depolymerization of tubulin to prevent cell division, and to inhibit the activity of DNA topoisomerase. Its glycoside derivatives etoposide and teniposide have low toxicity and have good curative effects on various tumor diseases such as small cell lung cancer and lymphoma, and have become a broad-spectrum anti-tumor drug in clinical application. [0003] At present, podophylloto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P19/60C12R1/66
Inventor 赵长琦贾栗朱妍妍艾嫦
Owner BEIJING NORMAL UNIVERSITY
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