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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  • Abstract
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  • 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 over

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0019] Example 1: Biosynthesis of GAMG

[0020] Seed medium: each liter of medium contains 5g glycyrrhizic acid, 3g 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℃ for 20min;

[0021] Fermentation culture: each liter of medium contains 5g 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℃ for 20min;

[0022] Transformation solution: 10g glycyrrhizic acid, 0.05M pH 7.0 phosphate buffer at a constant volume of 500ml.

[0023] The slant strain producing Penicillium violaceum was connected to the seed culture medium, and cultured in a shaker at 30°C for 24h at 180r / min. The seed liquid was transferred to the fermentation medium with a volume ratio of 1:10 inoculum, cultured on a shaker at 28°C and 180r / min for 96 hours to obtain a fermentation broth. The fermentation broth was added to the conversion broth at a ratio of 50%. The concentration of glycyrrhiz...

Example Embodiment

[0024] Example 2: Biosynthesis of GAMG

[0025] Seed medium: 0.5g glycyrrhizic acid, 1g 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, distilled water 1000mL, 121℃ sterilized 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, 121℃ sterilized for 20min;

[0027] Transformation solution: 8g glycyrrhizic acid, 0.05M pH 7.0 phosphate buffer solution with a constant volume of 500ml.

[0028] The penicillium violaceum-producing slant strain was connected to the seed culture medium, and cultured on a shaker at 25°C for 20 hours at 100 r / min. The seed liquid is transferred to the fermentation medium at a volume ratio of 1:10 inoculum, cultured in a shaker at 25°C and 100r / min for 72 hours to obtain a fermentation broth. The fermentation broth is added to the conversion broth at a ratio of 30%. The concentration of glycyrrhizic ...

Example Embodiment

[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, 121°C sterilization 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, 121°C sterilization for 20min;

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

[0033] The slant strain producing Penicillium violaceum was connected to the seed culture medium, and cultured on a shaker at 40°C for 24h at 300r / min. The seed liquid was transferred to the fermentation medium with a volume ratio of 1:10 inoculum. The fermentation broth was obtained by culturing at 40°C and 150r / min in a shaker for 96 hours. The fermentation broth was added to the conversion broth at a ratio of 50%. The concentration of glycy...

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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

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Claims

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Application Information

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Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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