High-yield antrodia cinnamomea mycelium fermentation method for triterpenoids
A fermentation method and technology of triterpenoids, applied in the field of fermentation of Antrodia camphorata mycelium with high yield of triterpenoids, can solve the problems of low yield of triterpenoids, achieve shortening of fermentation time, increase of yield, and increase of yield Effect
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[0033] 1. Preparation of oleanolic acid standard substance: Accurately weigh 10 mg of oleanolic acid standard substance, and use absolute ethanol as solvent to prepare 100 mL of oleanolic acid ethanol solution with a concentration of 0.1 mg / mL.
[0034] 2. Preparation of vanillin glacial acetic acid solution: Accurately and quickly weigh 0.552g of vanillin, quickly dissolve it with an appropriate amount of glacial acetic acid and immediately pour it into a 10mL volumetric flask, and quickly dilute it to the mark with glacial acetic acid for use on the day.
[0035] 3. Draw the standard curve: Accurately draw 0.1, 0.2, 0.3, 0.3, 0.4, 0.5mL of oleanolic acid standard solution and place them in a stoppered test tube, heat to evaporate the solvent, and then add 0.4mL of newly prepared 5% vanillin- Glacial acetic acid solution and 1.5mL perchloric acid were heated in a constant temperature water bath at 70°C for 15 minutes, cooled to room temperature with running water, then adde...
Embodiment 1
[0039] Embodiment 1 High yield triterpenoids Antrodia camphorata mycelia fermentation method
[0040] (1) Activation of slant strains
[0041] Transfer the slant strains of Antrodia camphorata (ATCC200183) to the EMA plate medium for activation and cultivation of the slant strains. When the colony diameter of Antrodia camphorata is greater than 2cm, transfer to fresh EMA plate medium for culture again, at 25°C, After culturing for 18 days, the mycelia covered the plate and formed orange-yellow spores.
[0042] (2) Shake flask liquid seed preparation
[0043] Spores of Antrodia camphorata were aseptically collected and placed in shake flasks for liquid culture at a temperature of 25°C and a rotational speed of 100 rpm for 6 days. The seed liquid appeared turbid but no obvious hyphae balls were formed.
[0044] Liquid seed medium composition: glucose 10g / L, malt extract 10g / L, peptone 1.0g / L, magnesium sulfate 0.5g / L, pH 5.0, distilled water to volume, 121°C, autoclaved for 20...
Embodiment 2
[0047] The mycelia fermentation method of Antrodia camphorata of embodiment 2 high-yield triterpenoids
[0048] (1) Activation of slant strains
[0049] Transfer the slant strains of Antrodia camphorata (ATCC200183) to the EMA plate medium for activation and cultivation of the slant strains. When the colony diameter of Antrodia camphorata is greater than 2cm, transfer to fresh EMA plate medium for culture again, at 28°C, After culturing for 15 days, the mycelia covered the plate and formed orange-yellow spores.
[0050] (2) Shake flask liquid seed preparation
[0051] Liquid seed medium composition: glucose 20.0g / L, malt extract 20.0g / L, peptone 5.0g / L, magnesium sulfate 0.5g / L, pH 4.0, distilled water to volume, 121°C, high-pressure steam sterilization for 20min.
[0052] Spores of Antrodia camphorata were aseptically collected and placed in shake flasks for liquid culture at a culture temperature of 28°C and a rotational speed of 120 rpm for 5 days. The seed liquid appeare...
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