Application of tremella fuciformis thalli in preparation of ginkgolide B
A technology of ginkgolide and tremella fungus, which is applied in the field of bioengineering and can solve problems such as difficulty in mass production of ginkgolide and the like
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Embodiment 1
[0014] The preparation of embodiment 1 test tube bacterial classification
[0015] Preparation of Potato Dextrose Agar Medium: Weigh 200g of potatoes, cut them into slices, add 1000mL of water, boil for 30 minutes, filter, dilute the filtrate to 1000mL, add 20g of agar, heat until the agar is completely melted, and pack into 18×200mm 15mL medium per tube, sterilized at 120°C for 30 minutes, cooled to 50°C, placed on a slope, the length of the slope is half the length of the test tube; cut the preserved Tremella fungus isolated from the fruiting body of Tremella into 4× 4mm small block bacteria, pick a small piece and transfer it to a test tube containing potato dextrose agar medium, culture at 20°C for 20 days, and prepare the test tube slant strain, and store the test tube slant at 4°C for later use;
Embodiment 2
[0016] Embodiment 2 contains the preparation of the fermented liquid of ginkgolide B
[0017] 1. Preparation of strains in liquid shake flasks
[0018] The formula of liquid shake flask culture medium is (in g / L) glucose 5, corn flour 5, peptone 1, KH 2 PO 4 1. MgSO 4 1. The initial pH value is 5.0. A total of 5500mL was prepared, divided into 250mL Erlenmeyer flasks, each bottle was 60mL, a total of 84 bottles were sterilized at 100°C for 60 minutes. A total of 5500mL was prepared, divided into 250mL triangular flasks, each bottle was 60mL, a total of 84 bottles, the preserved Tremella test tube slant strain isolated from the fruiting body of Tremella yunnanensis was cut into small pieces of 4×4mm size, and 3 pieces were picked for transfer Put it into a 250mL Erlenmeyer flask filled with shaker culture medium, and place it on a shaker at 22°C and 90 rpm for 24 hours.
[0019] 2. Preparation of seed tank strains
[0020] The 75L seed tank is filled with 45L seed culture...
Embodiment 3
[0025] Embodiment 3 contains the preparation of the fermented liquid of ginkgolide B
[0026] 1. Production of strains in shake flasks
[0027] The formula of shake flask culture medium is (unit: g / L): glucose 20, corn flour 15, peptone 6, KH 2 PO 4 3. MgSO 4 2. The initial pH value is 6.5, a total of 4500mL is prepared, divided into 250mL Erlenmeyer flasks (total 50 bottles), each bottle is 90mL, and sterilized at 120°C for 40 minutes. After cooling, cut the preserved tremella test tube slant strains separated from Shanxi tremella fruiting bodies into small pieces of 4×4mm size, and cut the test tube slant strains into small pieces of 4×4mm size, pick 4 pieces and transfer them into Place the Erlenmeyer flask with shaker culture medium on a shaker at 25°C and 120 rpm for 36 hours.
[0028] 2. Preparation of seed tank strains
[0029] 100L seed tank is equipped with 50L seed culture medium, wherein the formula of the seed tank culture medium is (unit is g / L): glucose 20, ...
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