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
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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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