Method for preparing rare ginsenoside CK by fermenting protopanoxadiol with penicillium adametzii
A technology of protopanaxadiol and adapenicillium, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc.
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Embodiment 1
[0018] (1) Cultivation of microorganisms: inoculate Penicillium adabilis (3.3203) from the solid medium into the pre-prepared liquid potato medium, the liquid potato medium contains 1kg of glucose in 100L; place it in a fully automatic fermenter, The culture temperature is 24 degrees, the aeration ratio is 0.1% (v / v), the stirring rate is 360 rpm / min, and the fermentation culture is 36 hours;
[0019] (2) Dissolve protopanaxadiol in methanol to make 100g / L, take 3L of the substrate drug solution and add it to the fermentation broth prepared in step (1), culture at 24°C, stir at 360rpm / min, and continue to ferment and cultivate 120 hours;
[0020] (3) Separation and purification of transformed products: collect the filtrate after centrifugation and filtration of the fermentation broth, concentrate under reduced pressure to 1 / 10 of the original volume, separate by HP20 macroporous resin column chromatography, and elute with deionized water to 2 times the column retentio...
Embodiment 2
[0025] (1) Cultivation of microorganisms: inoculate Penicillium adabilis (3.3203) from the solid medium into the pre-prepared liquid potato medium, and the liquid potato medium 100L contains 5kg of glucose; put it in a fully automatic fermenter, The culture temperature is 32 degrees, the aeration ratio is 10% (v / v), the stirring rate is 100 rpm / min, and the fermentation culture is 72 hours;
[0026] (2) Dissolve protopanaxadiol in 50% ethanol water solution by volume to prepare 10g / L, take 3L of the substrate drug solution and add it to the fermentation broth prepared in step (1), culture at 32°C, and stir at 100rpm / min, continue to ferment and cultivate for 48 hours;
[0027] (3) Separation and purification of transformed products: collect the filtrate after centrifugation and filtration of the fermentation broth, concentrate under reduced pressure to 1 / 20 of the original volume, separate by D101 macroporous resin column chromatography, and elute with deionized wate...
Embodiment 3
[0029] (1) Culture of microorganisms: inoculate Penicillium adabilis (3.3203) from the solid medium into the pre-prepared liquid potato medium, the liquid potato medium contains 2kg of glucose in 100L; put it in a fully automatic fermenter, The culture temperature is 28 degrees, the aeration ratio is 5% (v / v), the stirring rate is 220rpm / min, and the fermentation culture is 48 hours;
[0030] (2) Dissolve protopanaxadiol in methanol aqueous solution with a concentration of 50% by volume to make 50mg / ml, take 3L of the substrate solution and add it to the fermentation broth prepared in step (1), culture at 28 degrees, and stir at For 220rpm / min, continue to ferment and cultivate for 96 hours;
[0031] (3) Separation and purification of the transformed product: collect the filtrate after centrifugation and filtration of the fermentation broth, concentrate it under reduced pressure to 1 / 15 of the original volume, separate it by AB-8 macroporous resin column chromatogra...
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