Aspergillus oryzae for converting ginsenoside Rg3 to produce Rh2, production method and application

A technology of ginsenoside and Aspergillus oryzae, applied in the field of Aspergillus oryzae, can solve the problems of limited drug efficacy and low water solubility, and achieve the effects of low cost, simple operation, and promotion of modernization and industrialization

Active Publication Date: 2014-08-06
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, ginsenoside Rg 3 and Rh 2 Both have very important medicinal value, compared to ginsenoside Rh 2 , in practice due to Rg 3 The water solubility is low, which limits the efficacy to a certain extent

Method used

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  • Aspergillus oryzae for converting ginsenoside Rg3 to produce Rh2, production method and application
  • Aspergillus oryzae for converting ginsenoside Rg3 to produce Rh2, production method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0024] 1. Screening of Aspergillus oryzae

[0025] Take Daqu from a certain winery, and prepare 10 times with sterile saline -2 、10 -3 、10 -4 、10 -5 、10 -6 For suspensions with different dilution concentrations, draw 0.2 mL of suspensions with different concentrations and drop them on the surface of PDA medium added with chloramphenicol, spread evenly, and place the plate upside down and place it in a constant temperature incubator at 30°C for cultivation. After the colonies grew out, single colonies were picked and further purified. Focusing on the appearance of the colony, observe the shape of the bacteria with a microscope, and obtain a series of purely cultured filamentous fungal strains after multiple isolation and purification.

[0026] The three strains obtained by separation and purification were named DQ-MJ01, DQ-MJ02 and DQ-MJ03, and the ginsenoside conversion experiments were carried out respectively. The results showed that the strain DQ-MJ03 had a higher conv...

Embodiment 2

[0035] Aspergillus oryzae of the present embodiment transforms ginsenoside Rg 3 produce Rh 2 method, including the following steps:

[0036] (1) Preparation of strains: Inoculate Aspergillus oryzae AS.mq01 on PDA slant medium, culture at 36°C for 40 hours, then pick spores or colonies of the strain and transfer them to wort medium, and ferment at 28°C and 160rpm for 3 sky;

[0037](2) Preparation of enzyme liquid: filter the fermentation liquid in step (1), centrifuge the filtrate at 10,000 rpm for 20 min at 15°C, remove bacterial precipitates, and collect the supernatant; add ammonium sulfate to the supernatant to 70% saturation temperature; stand overnight at 4°C, centrifuge at 10,000rpm for 20min at 15°C, discard the supernatant, and collect the protein precipitate; dissolve the protein precipitate with 0.02mol / L, pH 5.0 acetic acid-sodium acetate buffer solution and dialyze Dialyze the bag with the same acetic acid-sodium acetate buffer solution for 24 hours; after dial...

Embodiment 3

[0041] Aspergillus oryzae of the present embodiment transforms ginsenoside Rg 3 produce Rh 2 method, including the following steps:

[0042] (1) Preparation of strains: inoculate Aspergillus oryzae AS.mq01 on PDA slant medium, culture at 32°C for 48 hours, then pick the spores or colonies of the strain and transfer them to wort medium, and ferment at 32°C and 200rpm for 3 sky;

[0043] (2) Preparation of enzyme liquid: filter the fermentation liquid in step (1), centrifuge the filtrate at 13,000 rpm for 10 min at 20°C, remove bacterial precipitates, and collect the supernatant; add ammonium sulfate to the supernatant to 60% saturation temperature; stand at 6°C overnight, centrifuge at 13,000rpm for 10min at 20°C, discard the supernatant, and collect the protein precipitate; dissolve the protein precipitate with 0.02mol / L, pH 4.5 acetic acid-sodium acetate buffer solution and dialyze Dialyze the bag with the same acetic acid-sodium acetate buffer solution for 30 hours; after...

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Abstract

The invention discloses aspergillus oryzae for converting ginsenoside Rg3 to produce Rh2, a production method and an application. The aspergillus oryzae is named AS.mq01 and preserved in the CCTCC (China Center for Type Culture Collection) in Wuhan University located on Mountain lojia, Wuchang, Wuhan City, Hubei Province on September 17th, 2013 with the preservation number of CCTCC NO: M2013430. Aspergillus oryzae metabolites obtained through screening contain ginsenoside glycosidase capable of powerfully hydrolyzing the ginsenoside Rg3 to convert the ginsenoside Rg3 to produce the Rh2 within 24 h, the conversion rate is up to 56.6%, and in addition, the Rh2 produced through conversion cannot be further decomposed and converted.

Description

technical field [0001] The present invention relates to a kind of transformed ginsenoside Rg 3 produce Rh 2 of Aspergillus oryzae, also involved in the conversion of Aspergillus oryzae to ginsenoside Rg 3 produce Rh 2 The method and application of the invention belong to the technical field of traditional Chinese medicine. Background technique [0002] Ginseng (Panax ginseng G.A. Meyer) is a precious traditional Chinese medicine in my country, which has been used in Chinese medicine for more than two thousand years. The main active ingredient of ginseng is ginsenoside (ginsenoside), among which protopanaxadiol saponins belong to dammarane-type tetracyclic triterpene saponins, mainly including Rb1, Rc, Rd, Rg 3 and Rh 2 Wait. Among them, ginsenoside Rg 3 and Rh 2 Both have very important medicinal value, compared to ginsenoside Rh 2 , in practice due to Rg 3 The water solubility is low, which limits the efficacy to a certain extent. In addition, ginsenoside Rg 3 I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P33/20C12R1/69
Inventor 李东霄马金友莫海珍常景玲华承伟吴羽晨张蕾
Owner HENAN INST OF SCI & TECH
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