Strain of Dongxiang wild rice endophytic fungus for producing GAMG (glycyrrhetinic acid mono-glucuronide) by efficiently converting glycyrrhizic acid and application thereof
A technology of endophytic fungi and glycyrrhizic acid, applied in the direction of fungi, microorganism-based methods, microorganisms, etc., can solve many problems such as low energy consumption, high yield, and simple operation
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Embodiment 1
[0027] The separation of Chaetomium globosum DX-THS3 of the present invention:
[0028] (1) Collect complete Dongxiang wild rice and return it to the laboratory for immediate processing. Roots, stems and leaves are all soaked in 75% alcohol for 5 minutes for preliminary sterilization, then soaked in 0.1% mercury chloride for 8 minutes, and then rinsed with sterile water to sterilize the surface of the tissue.
[0029] (2) The above-mentioned treated stems are all subtracted with sterilized scissors, and then the stems with the two ends removed are cut longitudinally, divided into two, and then cut into small pieces of 0.1cm×0.5cm. They were respectively placed on PDA medium supplemented with 60 μg / L streptomycin and 0.5 g / L potassium dichromate, and cultured in an incubator at 28°C. In order to check the effect of surface sterilization, a control group was set up to test the effect of sterilization. Control Ⅰ: The sterilized roots, stems and leaves are not subtracted from th...
Embodiment 2
[0033] Screening of Chaetomium globosum DX-THS3 of the present invention:
[0034] (1) Plate initial screening: Activate the Dongxiang wild rice endophytic fungus obtained from the above separation and purification, and then inoculate it into a solid screening medium with glycyrrhizic acid (glycyrrhizinate or licorice extract) as the only carbon source, and the control group uses glucose as the only carbon source. Placed in an incubator at 28°C, cultured continuously for 7 days, and screened out the strains that can grow well on the medium with glycyrrhizic acid as the only carbon source.
[0035] The composition and proportioning of the solid screening medium are: ammonium glycyrrhizinate 1.0g / L, KH 2 PO 4 2.2g / L, NH 4 NO 3 3.0g / L, NaCl 0.5g / L, yeast powder 0.05g / L, MgSO 4 ·7H 2 O 0.25g / L, CaCl 2 0.011g / L, ZnSO 4 ·7H 2 O2.87mg / L, MnSO 4 ·H 2 O 1.69mg / L, H 3 BO 3 0.03mg / L, agar powder 20g / L, pure water 1000mL, adjust the pH to 5.0-7.0, and sterilize under high-...
Embodiment 3
[0041] Transformation of licorice extract by Chaetomium globosum DX-THS3 to produce GAMG:
[0042] Inoculate the activated Chaetomium globosum DX-THS3 into the seed medium, place it in a shaker with a temperature of 28°C and a rotation speed of 120r / min, and cultivate it continuously for 3 days, according to 1-10% (w / Inoculation amount of v) Inoculate the above-mentioned cultivated seeds into the transformation medium containing glycyrrhizic acid, place them in a shaker at a temperature of 28° C. and a rotation speed of 120 r / min, and cultivate continuously for 7 days. The bacterial cells were removed by centrifugation, the fermentation broth was collected, and a 0.22 μm microporous filter head was used to detect the transformation product by HPLC.
[0043] The composition and proportioning of described seed culture medium are: glucose 20g / L, KH 2 PO 4 1.2~3.2g / L, NH 4 NO 3 2.0~5.0g / L, NaCl 0.5~1.0g / L, yeast powder 0.05~0.3g / L, MgSO 4 ·7H 2 O 0.25~0.5g / L, CaCl 2 0.01...
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