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Method for oriented biosynthesis of GAMG

A biosynthesis and seed technology, applied in the biosynthesis of monoglucuronic acid glycyrrhetinic acid, and the field of directional catalysis of glycyrrhizic acid to synthesize GAMG, can solve the problems of poor catalysis orientation, easy environmental pollution, and low chemical synthesis selectivity, Achieve the effect of low cost, single product and high reaction efficiency

Inactive Publication Date: 2009-12-16
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of this invention is to provide a kind of method of directional biosynthesis monoglucuronyl glycyrrhetinic acid (GAMG), to overcome the low selectivity of chemical synthesis of existing GAMG, poor catalytic orientation and the deficiency that environment is easily polluted

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0019] Example 1: Biosynthesis of GAMG

[0020] Seed medium: Each liter of medium contains 5g of glycyrrhizic acid, 3g of glucose, NH 4 NO 3 3g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5g, KCl 0.5g, FeSO 4 ·7H 2 O 0.001g, sterilized at 121°C for 20min;

[0021] Fermentation culture: Each liter of medium contains 5g of glycyrrhizic acid, NH 4 NO 3 3g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5g, KCl 0.5g, FeSO 4 ·7H 2 O 0.001g, sterilized at 121°C for 20min;

[0022] Transformation solution: Glycyrrhizic acid 10g, 0.05M pH 7.0 phosphate buffer solution to volume 500ml.

[0023] The Penicillium purpura slant strains were inserted into the seed medium, and cultured on a shaker at 30°C for 24 hours at 180r / min. The seed liquid is transferred to the fermentation medium with the inoculation amount of 1:10 by volume ratio, 28 ° C, 180r / min shaker culture for 96 hours to obtain the fermentation liquid, the fermentation liquid is added into the transformation liquid in a ratio of 50%,...

example 2

[0024] Example 2: Biosynthesis of GAMG

[0025] Seed medium: glycyrrhizic acid 0.5g, glucose 1g, NH 4 NO 3 3g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5g, KCl 0.5g, FeSO 4 ·7H 2 O 0.001g, distilled water 1000mL, sterilized at 121°C for 20min;

[0026] Fermentation medium: glycyrrhizic acid 0.5g, NH 4 NO 3 3g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5g, KCl 0.5g, FeSO 4 ·7H 2 O 0.001g, distilled water 1000mL, sterilized at 121°C for 20min;

[0027] Transformation solution: Glycyrrhizic acid 8g, 0.05M pH 7.0 phosphate buffer solution to volume 500ml.

[0028] The Penicillium purpura slant strains were inoculated into the seed medium, cultured on a shaker at 25°C for 20 hours at 100 r / min. The seed liquid is transferred to the fermentation medium with the inoculum amount of 1:10 by volume ratio, 25 ° C, 100r / min shaker culture for 72 hours to obtain the fermentation liquid, the fermentation liquid is added into the transformation liquid in a ratio of 30%, the concentration of g...

example 3

[0029] Example 3: Biosynthesis of GAMG

[0030] Seed medium: glycyrrhizic acid 8g, glucose 5g, NH 4 NO 3 3g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O0.5g, KCl 0.5g, FeSO 4 ·7H 2 O 0.01g, 1000mL distilled water, sterilized at 121°C for 20min;

[0031] Fermentation medium: glycyrrhizic acid 8g, NH 4 NO 3 3g, KH 2 PO 4 1g, MgSO4 ·7H 2 O 0.5g, KCl 0.5g, FeSO 4 ·7H 2 O 0.01g, 1000mL distilled water, sterilized at 121°C for 20min;

[0032] Transformation solution: 40 g of glycyrrhizic acid, 500 ml of 0.05M pH 7.0 phosphate buffer.

[0033] The Penicillium purpurea slant strains were inoculated into the seed medium, cultured on a shaker at 40°C for 24 hours at 300 r / min. The seed liquid is transferred to the fermentation medium with an inoculation amount of 1:10 by volume ratio, 40 ° C, 150r / min shaker culture for 96 hours to obtain a fermentation liquid, and the fermentation liquid is added to the transformation liquid in a ratio of 50%. The concentration of glycyrrhizic aci...

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Abstract

The invention provides a method for oriented biosynthesis of GAMG, and belongs to the technical fields of biosynthesis and biotransformation. The method comprises the following steps: producing corresponding beta-D-glucuronidase by penicillium purpurogenum stoll in a fermentation medium; adding the obtained beta-D-glucuronidase into conversion solution containing glycyrrhizic acid substrate, orientedly hydrolyzing the glycyrrhizic acid under the action of enzyme, and obtaining a coarse product of GAMG; and performing organic solvent extraction, resin adsorption and purification to obtain a GAMG product. The method takes microbialenzyme as a catalyst, and has high reaction efficiency, no pollution, simple steps, and low cost; and the product is single and has high purity.

Description

technical field [0001] The invention relates to a biosynthetic method of monoglucuronyl glycyrrhetinic acid (GAMG), in particular to a method for directionally catalyzing glycyrrhizic acid to synthesize GAMG under the action of microbial enzymes, and belongs to the technical field of biosynthesis and biotransformation. Background technique [0002] Licorice is a leguminous plant of the genus Glycyrrhizae. Glycyrrhizic acid is one of the main active ingredients in licorice, which has anti-inflammatory, anti-virus, anti-tumor, and anti-digestive ulcer effects. In addition, glycyrrhizic acid is also an important natural sweetener, its sweetness is about 200 times that of sucrose, and its calories are low, so it is widely used as a food additive. [0003] Glycyrrhizin (GL) is one of the main active ingredients of the traditional Chinese medicine licorice. It consists of two glucuronic acids connected by pentacyclic triterpene saponins through β-1,2 glycosidic bonds and β-1,3 gl...

Claims

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Application Information

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IPC IPC(8): C12P19/56C12R1/80
Inventor 李春王小艳邹树平冯世江戴大章
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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