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Methods for screening biotransformation strain of resveratrol, identifying strain and biotransforming resveratrol by using strain

A resveratrol and biotransformation technology, applied in the field of biotransformation of plant active substances, can solve problems such as large pollution, high synthesis cost, and poor product safety

Inactive Publication Date: 2012-10-03
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the method of chemically synthesizing resveratrol has made great progress, all methods still have problems such as high synthesis cost, large pollution, and poor product safety, which need further improvement

Method used

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  • Methods for screening biotransformation strain of resveratrol, identifying strain and biotransforming resveratrol by using strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: the screening method of resveratrol biotransformation bacterial classification, concrete steps are:

[0026] (1) Resveratrol glycoside pretreatment and medium configuration

[0027] Add 0.5g of resveratrol glycosides to 10mL of sterile ethanol in a sterile working environment, and evaporate to dry naturally to obtain sterile resveratrol glycosides for use;

[0028] Weigh 0.3g of sodium nitrate, 0.1g of dipotassium hydrogen phosphate, magnesium sulfate (MgSO 4 ·7H 2 O) 0.05g, potassium chloride 0.05g, ferrous sulfate 0.001g in a volumetric flask, and distilled water to 100mL to obtain the basal medium. Before use, weigh 100 mg of sterile resveratrol glycosides and add it to the sterilized basal medium to prepare an enriched medium.

[0029] (2) Similarly, use the above-mentioned basal medium and add 1.5% agar by weight of the basal medium to fix it, heat to dissolve before use, weigh 100 mg of sterile resveratrol glycosides and add it when it is cooled t...

Embodiment 2

[0036] Example 2: Identification method of resveratrol biotransformation strains.

[0037] The morphological observation and molecular biological identification of the strain S-4 to be identified were carried out respectively. The specific methods are:

[0038] 1) For the morphological observation of the strains, culture the PDA as a solid, pour the Chase solid medium onto the plate, pick up a little mycelium with an inoculation hook after condensation, spot it on the plate, and culture it upside down in a 30°C incubator for more than 48 hours, and observe the colonies The process and form of formation;

[0039] 2) Prepare slices by soaking in water, add a drop of lactic acid carbolic acid blue cotton solution to the center of a clean slide, use a dissecting needle to draw a small piece of mycelium from the edge of the colony, put it into the lactic acid carbolic acid blue cotton solution, and cover it. slides, observed under a microscope. Observe whether there are diaphragm...

Embodiment 3

[0051] The method for converting resveratrol analogs, the steps are:

[0052] 1) Take 10.0 g of broken Polygonum cuspidatum and put it in a beaker, add 20 mL of deionized into it, let it stand for 16-24 hours, and obtain the Polygonum cuspidatum leachate for later use;

[0053] 2) Put another PDA medium into the Erlenmeyer flask, add 20 μL of Polygonum cuspidatum extract, and seal it for later use;

[0054] 3) Add 0.5 mL of the bacterial liquid to 1 mL of the polydatin solution obtained in 2), seal and incubate at 28°C, protected from light, and shake at 200 r / min for 4 days;

[0055] 4) After the culture solution was diluted five times with methanol, the conversion rate was analyzed by high performance liquid chromatography (chromatographic conditions: Dima C18 diamond column (200 × 4.6mm); detection wavelength: 306nm; mobile phase: 0.05% phosphoric acid aqueous solution: acetonitrile = 65:35; flow rate: 1 mL / min; injection volume 10 μL.). The results showed that 80 μg / mL r...

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Abstract

The invention discloses methods for screening a biotransformation strain of resveratrol, identifying the strain and biotransforming the resveratrol. The method for screening the biotransformation strain of the resveratrol comprises the following steps of: adding an enrichment culture medium into Japanese knotweed leaching liquor, sealing, performing shake culture at the rotating speed of 200r / min at the temperature of 28 DEG C for 4 days in a dark place, performing gradient dilution plate coating on a culture solution through which highest transformation efficiency is obtained on a screening culture medium, culturing at the temperature of 28 DEG C, taking a hydrolysis circle culture medium, extracting by using methanol, and performing thin-layer chromatography (TLC) analysis and high performance liquid chromatography (HPLC) analysis to obtain a strain S-4 with high transformation capacity. Through the identification, the strain is Penicillium camembert. The culture solution of the strain has strong catalytic capacity and can completely transform the polydatin into the resveratrol; and meanwhile, generated glucose is used as a carbon source of the strain, and strains which do not have catalytic capacity can die slowly in the growth process. The methods have an important significance for the high-efficiency, safe and low-cost production of the resveratrol.

Description

technical field [0001] The invention relates to the technical field of biotransformation of plant active substances, in particular to a screening method and identification of strains capable of efficiently transforming resveratrol analogues into resveratrol and using the strains to perform resveratrol Methods of biotransformation. Background technique [0002] As a natural active substance, resveratrol has a low content in nature, while resveratrol and glucose-bound glycosides—resveratrol glycosides, have a relatively high content. Research on the method of effectively increasing the production of resveratrol The approach has important industrial significance. Du Jianli and others established a chemical synthesis method, the total yield of resveratrol was 41.9%, and resveratrol was successfully synthesized in only three steps (Du Jianli, Zou Yong, Xiao Chunfen. The simple synthesis of resveratrol. Fine Chemical Industry, 2009, 26(6): 580-584). Ding Liugang et al. improved...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N1/02C12Q1/68C12Q1/04C12P7/22C12R1/80
Inventor 陈雪香曹庸李赟
Owner SOUTH CHINA AGRI UNIV
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