Bacillus Velez g9y and its application in ginsenoside biotransformation

A technology of ginsenoside and bacillus, applied in the field of new high-efficiency ginsenoside transformation bacteria, can solve the problems of long fermentation transformation period, increased purification, difficulties, etc., and achieve the effects of short fermentation transformation time, easy extraction and purification, and low cost

Active Publication Date: 2022-04-01
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention patent application document with the publication number CN104140931A discloses a method for converting and synthesizing ginsenoside CK by using PeniciIlium adametzii 3.3203; The method of ginsenoside Rd, this method adopts Coniochaetasp.EA-9 or its secreted crude enzyme solution to biotransform the total saponins of Panax notoginseng to obtain ginsenoside Rd and C-K; the product of this method is not single Ginsenoside Rd increases the difficulty for subsequent purification, and the fermentation conversion period is longer (6-12d)

Method used

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  • Bacillus Velez g9y and its application in ginsenoside biotransformation
  • Bacillus Velez g9y and its application in ginsenoside biotransformation
  • Bacillus Velez g9y and its application in ginsenoside biotransformation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: screening and identification of bacterial strain

[0046] 1) Medium:

[0047] Beef extract peptone (NA) solid medium: NaCl 5.0g / L, beef extract 3.0g / L, peptone 10.0g / L, agar 18g / L, dissolved in distilled water, autoclaved at 121°C for 20min, stored at 4°C for later use ;

[0048] Beef extract-peptone (NA) liquid medium: NaCl 5.0g / L, beef extract 3.0g / L, peptone 10.0g / L, dissolved in distilled water, autoclaved at 121°C for 20min, and stored at 4°C for later use.

[0049] 2) Isolation of strains

[0050] Pick the disease-free American ginseng (root), rinse the soil on the surface with flowing water and transfer it to the ultra-clean workbench. After rinsing with sterile water several times, use sterile filter paper to remove surface moisture, disinfect with alcohol (70% v / v) for 3 minutes, then immerse in sodium hypochlorite solution (4% w / v) for 5 minutes, and then rinse thoroughly with sterile water, Take 100ul of sterile water for the last rinse as a...

Embodiment 2

[0058] Embodiment 2: the preparation of Bacillus veleisi G9y bacterium liquid and the conversion of ginsenoside Rc→Rd

[0059] (1) Slant culture: Inoculate strain G9y on NA slant medium, culture at 37°C for 36 hours, and obtain bacterial slant;

[0060] (2) Seed culture: pick bacteria from the slope of the bacteria in step (1) and inoculate them into NA liquid medium, and cultivate them at 37°C and pH 6 for 16 hours to obtain G9y seed liquid;

[0061] (3) Expanded cultivation: inoculate the seed solution obtained in step (2) into NA liquid medium with an inoculum volume concentration of 5%, and cultivate it at 37° C. and pH 7 for 4 hours to obtain a G9y culture solution, which is used as a G9y bacterial solution For the fermentation transformation of ginsenoside Rc;

[0062] (4) Fermentation conversion: ginsenoside Rc is uniformly mixed with the G9y nutrient solution of step (3), in the fermented solution where Rc is mixed with the G9y bacterium solution, the initial ginsenos...

Embodiment 3

[0063] Example 3: Preparation of Bacillus Velez G9y bacterium liquid and transformation of saponins in ginseng powder

[0064] (1) Slant culture: Inoculate the strain G9y on a conventional LB slant medium, culture at 37°C for 36 hours, and obtain a bacterial slant;

[0065] (2) Seed culture: pick the bacteria from the slant of the bacteria in step (1) and inoculate them into a conventional LB liquid medium, cultivate at 37°C and pH 6 for 36 hours, and prepare the G9y seed liquid;

[0066] (3) Expanded cultivation: inoculate the seed solution obtained in step (2) into a conventional LB liquid medium with an inoculum volume concentration of 10%, and cultivate at 37° C. and pH 7 for 16 hours to obtain a G9y culture solution;

[0067] (4) Bacteria purification: Centrifuge the G9y culture solution at 6000rpm for 10min, discard the supernatant, and suspend the precipitate with 0.1mol / L, pH value of 7.0 phosphate buffer to prepare the G9y bacteria solution, which is used for the prod...

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Abstract

The present invention provides a new strain of high-efficiency ginsenoside transformation bacteria——Bacillus velezensis sp. G9y (preserved in China Center for Type Culture Collection, numbered CCTCC No: M 2019837, date of preservation is October 2019 17th) and its application in ginsenoside biotransformation. The Bacillus velezensis sp. G9y cultivation and fermentation process of the present invention is simple and low in cost; the G9y bacterial liquid is mixed with ginsenoside Rc or ginsenoside raw materials containing Rc, and the ginseng can be selectively, rapidly and efficiently transformed into ginseng by fermentation Saponin Rc → Rd, and the product is a single ginsenoside Rd, which is convenient for extraction and purification; the required fermentation conversion time is short (16-24h), and the conversion speed is much higher than that of the prior art (6-12d), which can greatly improve production efficiency and has great advantages. Good application prospects.

Description

technical field [0001] The invention relates to a novel high-efficiency ginsenoside transforming bacterium—Bacillus velesi G9y and its application in ginsenoside biotransformation. Background technique [0002] Ginsenoside is the most important medicinal active ingredient of Panax genus plants (ginseng, Panax notoginseng, American ginseng, etc.), which has high pharmacological activity and medicinal value. According to the structure of ginsenogenin, it can be divided into three categories: protopanaxadiol, protopanaxatriol and oleanolic acid. Ginsenoside Rd is a protopanaxadiol saponin. Compared with other protopanaxaponins (Rb1, Rb2, Rc), its sugar chain is shorter and easier to be absorbed by the human intestine. Extensive studies have shown that ginsenoside Rd has protective functions on the nervous system, such as repairing nerve gas or ischemic stroke caused by cell damage, promoting the differentiation of neural stem cells into astrocytes, and increasing the vitality ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P33/20C12R1/07
CPCC12N1/20C12P33/20C12R2001/07C12N1/205
Inventor 朱大恒张臣许妍妍朱润琪
Owner ZHENGZHOU UNIV
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