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Polygonum cuspidatum endogenous bacterial strain aspergillus fumigates J3 bacterial strain for converting polydatin

A technology of Polygoniside and Aspergillus fumigatus, which is applied to microorganisms, fungi, microorganisms and other directions, can solve the problems of polluted environment, many by-products of chemical methods, and complicated reaction steps.

Inactive Publication Date: 2015-07-15
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to obtain active derivatives of polydatin with novel structures, the commonly used methods include chemical synthesis or modification of its structure and microbial transformation, but chemical methods have many by-products, cumbersome reaction steps, and pollute the environment, while microbial transformation has regio and stereoselectivity Strong, simple operation, mild reaction conditions, low cost, etc., can complete some reactions that are difficult to achieve by chemical methods

Method used

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  • Polygonum cuspidatum endogenous bacterial strain aspergillus fumigates J3 bacterial strain for converting polydatin
  • Polygonum cuspidatum endogenous bacterial strain aspergillus fumigates J3 bacterial strain for converting polydatin

Examples

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Embodiment 1

[0020] Example 1 Preparation of strain J3 of the present invention

[0021] Take wild Polygonum cuspidatum stalks collected from Lianyuan, Hunan Province, rinse with sterile water, then absorb the water with sterile filter paper, soak in 75% ethanol for 3-5 min, rinse with sterile water 3-4 times, 0.1% liter Sterilize with mercury for 30 seconds, and then use 75% ethanol for 1 minute. Cut off the root and stem phloem, take the xylem and cut it into small pieces of 0.5cm×0.5cm with sterile scissors. Rinse 3-4 times with sterile water and grind into a slurry in a sterilized mortar. After the grinding slurry is diluted 10-fold with sterile water, 1 ml of the grinding diluent is removed, and routinely spread on the primary screening medium. Each material was cultured three times in parallel at 28℃ for 72-120h, and the well-growing colonies with different morphologies were selected and streaked on a PDA plate. After 72h at 28℃, a single colony was collected and streaked 3 times to o...

Embodiment 2

[0022] Example 2 Preparation of fermentation broth of strain J3 of the present invention

[0023] The strain J3 of the present invention was inoculated into the PDA slant medium for activation at 28°C for 72 hours, and then 20% of the inoculum amount was connected to the transformation medium, that is, the PDB medium containing the substrate 10 mg polydatin (PDB medium: 200 g potato, glucose In 20 g, 1000 ml of water, natural pH), the fermentation broth was prepared by biotransformation at 28° C. and 180 r / min for 96 hours.

Embodiment 3

[0024] Example 3 Separation and purification of biotransformation products of fermentation broth of strain J3 of the present invention

[0025] Centrifuge the fermentation broth prepared above at 10,000 r / min for 10 min to remove mycelium and impurities, and collect the supernatant. Take 3L of the supernatant, absorb it with 300g AB-8 macroporous resin, and desorb it with 30%, 50%, 70%, 90%, and absolute ethanol, until the effluent of each stage is colorless, collect them separately, and use chloroform: methanol It is 5:1 as the developing agent, and the conversion product is detected according to TLC tracking (with the known 6 standard products of polydatin, resveratrol, α-glucin, ε-glucose, pterostilbene and oxidized resveratrol TLC developed After the development point with different Rf value is the target component), combine the collected liquids with the same Rf value and concentrate; separate the concentrated solution by preparative thin layer chromatography (PTLC), develop...

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Abstract

The invention relates to olygonum cuspidatum endogenous bacterial strain for converting polydatin. The olygonum cuspidatum endogenous bacterial strain is classified and named as aspergillus fumigates J3 (Aspergillus fumigates ) bacterial strain. The olygonum cuspidatum endogenous bacterial strain had been preserved in China Center for Type Culture Collection in March 25, 2015, and the collecting umber is CCTCC M2015161. Biotransformation fermental cultivation is conducted on the bacterial strain in a PDB culture medium with 10 mg substrate polydatin. A biotransformation product component B possesses ability of eliminating DPPH. free radical and OH free radical in vitro after being separated and purified, and the capability to elimite OH free radical of the biotransformation product component B is greater than that of substrate polydatin. In this way, the bacterial strain J3 can conduct biotransformation on polydatin, ramification with higher antioxidant activity is formed, and more lead compound is provided for screening of natural antioxidant activity drug.

Description

Technical field [0001] The invention relates to an endophyte of Polygonum cuspidatum, in particular to a strain of Polygonum cuspidatum endophyte J3 that can transform polydatin to form an antioxidant active substance. Background technique [0002] Polygonum cuspidatum is a perennial herbaceous plant belonging to the genus Polygonaceae, and its dry rhizome is rich in polydatin, the content of which can reach 2.55%. The chemical name of polydatin is 3,4'-5-trihydroxystilbene-3-β-D-glucoside, which is a stilbene polyphenol substance, which can protect the cardiovascular system, resist thrombosis, lower blood lipids, and relieve cough Various pharmacological effects such as asthma, anti-oxidation and anti-pathogenic microorganisms. In plants, polydatin is mainly a stable trans isomer, and the sugar group is the main substituent. In recent years, research has found that the hydrogen on the two hydroxyl groups, sugar groups or benzene ring of polydatin is replaced by methyl or isopen...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P1/02C12R1/68
CPCC12P1/02C12N1/145C12R2001/68
Inventor 易有金罗程印隆丽林柏连阳
Owner HUNAN AGRICULTURAL UNIV
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