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Biotransformation method of rare ginsenoside Rg3

A technology of rare ginseng saponins and biotransformation, applied in the direction of fermentation, can solve the problems of high cost, low conversion efficiency, long time consumption, etc., and achieve the effects of high conversion efficiency, improved conversion efficiency and environmental friendliness

Active Publication Date: 2019-01-11
NORTHEAST FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing preparation methods of rare ginseng saponin Rg3 have defects such as low conversion efficiency, long time consumption, and high cost to varying degrees, which need to be improved urgently

Method used

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  • Biotransformation method of rare ginsenoside Rg3
  • Biotransformation method of rare ginsenoside Rg3
  • Biotransformation method of rare ginsenoside Rg3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Prokaryotic expression and functional verification of β-glucosidase BglPm, α-L-arabinofuranosidase Abf22-3, β-glucosidase Bgp1 and α-L-arabinopyranosidase Bgp2

[0033] 1. Construction and induced expression of prokaryotic expression vectors of BglPm, Abf22-3, Bgp1, Bgp2

[0034] 1.1 Construction of expression vector

[0035] According to the sequence of these four enzymes provided by Genbank, the base sequence is optimized, which is more conducive to the induction of prokaryotic expression. The optimized sequence is added with the cleavage site of the endonuclease KpnI / BamHI 5'-3'GGTACC / 5'- After 3'GGATCC, gene synthesis is carried out. Since the Bgp1 nucleotide sequence contains the restriction site of KpnI / BamHI, the restriction site of endonuclease NdeI / XmaI is added at 5'-3'CATATG / 5'-3 ' CCCGGG, see the sequence listing for the optimized gene sequence.

[0036] The synthesized gene sequence was subjected to double enzyme digestion at 37°C for 2 hours, ...

Embodiment 2

[0045] Example 2 Enzyme Combination Transformation of Total Ginsenosides to Generate Rg3

[0046] 1 Preparation of ginseng total saponins stock solution

[0047] (1) Take by weighing 15g of ginseng adventitious root after drying, put it into a mortar and grind until it becomes powdery, stop grinding and put it into a 50mL centrifuge tube;

[0048] (2) Add 25mL of 80% methanol into a 50mL centrifuge tube, put it into an ultrasonic cleaner for 1 hour, shake once every 10 minutes, and take the supernatant after the end; repeat the above steps once.

[0049] (3) Centrifuge the extract obtained in the previous step at room temperature for 10 min at 3000 rpm.

[0050] (4) filter to get the filtrate.

[0051] (5) The obtained filtrate was evaporated to dryness in a rotary evaporator.

[0052] (6) Add 10 mL of 80% methanol to redissolve.

[0053] 2 Purification of Ginseng Total Saponins Extract

[0054] (1) Soak 50mL D101 macroporous resin in 95% ethanol for 24 hours, wash with w...

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PUM

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Abstract

The invention discloses a biotransformation method of rare ginsenoside Rg3, comprising the steps: (1) extracting ginseng to obtain total ginsenoside extract; (2) adding any one or more of Beta-Glucosidase BglPm, a-L-Arabinofuranase Abf22-3, Beta-Glucosidase Bgp1 or a-L-arabinopyranosidase Bgp2 in the total ginsenoside extract for carrying out biotransformation to obtain a transformation product containing rare ginsenoside Rg3. The invention adds any one or more enzymes of Beta-Glucosidase, a-L-Arabinofuranase, Beta-Glucosidase or a-L-arabinopyranosidase in the total ginsenoside extract for biotransformation, and the biotransformation efficiency of the rare ginsenoside Rg3 is remarkably improved. The biotransformation method of the invention is environmentally friendly, has high conversionefficiency and is suitable for large-scale production and application.

Description

technical field [0001] The invention relates to a preparation method of rare ginseng saponin Rg3, in particular to a biotransformation method of rare ginseng saponin Rg3, and belongs to the field of preparation of rare ginseng saponin Rg3. Background technique [0002] Rg3 is a tetracyclic triterpene saponin, its molecular structure is as follows figure 2 As shown, the molecular formula is C 42 h 72 o 13 , with a molecular weight of 784.3, which was first prepared and determined by Japanese scholar Isao Kitagawa in 1980, and it was proposed that ginsenoside Rg3 has the effect of selectively inhibiting tumor cell infiltration and metastasis. Among the more than 180 saponins that have been discovered, ginsenoside monomers Rb1, Rb2, Rc, etc. account for more than 50% of the total saponins, while Rg3, which is called a rare ginsenoside, only accounts for 0.015% of the dry weight of ginseng. [0003] The existing preparation methods of rare ginseng saponin Rg3 have defects su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/06
CPCC12P33/06
Inventor 王宇李玉花吴昊曹领改尚艳茹李方远
Owner NORTHEAST FORESTRY UNIVERSITY
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