Industrial preparation process for producing paclitaxel by fermenting taxus endophytic fungus mutant
A technology of endophytic fungi and preparation technology, which is applied in the field of bio-metabolism synthesis and purification, can solve the problems of no industrial production of paclitaxel, no prospect of industrial application, low yield of paclitaxel compounds, etc., and achieve large production capacity, low product cost, The effect of optimizing energy consumption
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[0102]The preparation and industrial fermentation method of bacterial classification are as follows:
[0103] i. The formula of liquid shake flask culture medium is PDA, pH 7, initial pH value is 5.0, a total of 250ml is prepared, divided into 250mL triangular flasks, each bottle is 30ml, sterilized at 121°C for 30 minutes, then cooled to room temperature for use;
[0104] ii. Pick and transfer the cultured strains on the slant of the test tube into a 250ml Erlenmeyer flask equipped with a shaker culture medium, and place it in a shaking table at 28°C and 120 rpm for 24 hours;
[0105] iii. Prepare the seed culture medium according to the formula, the formula of the described seed culture medium is 40g of sucrose, 10g of peptone, 10g of yeast extract powder, 15g of agar, 1000mL of distilled water, pH 7; the seed culture medium is pumped into No. 1-8 500L- Grade seed culture tanks, sterilized by steam sterilization in real tanks, with a sterilization temperature of ≥121°C and a...
Embodiment 1
[0162] The isolation, mutagenesis and preservation method steps of Trichoderma HQ-24 bacterial strain are as follows:
[0163] a. Using the conventional tissue separation method, cut the fresh Taxus chinensis fruit in Yunnan area into 0.5-1cm 2 small pieces. Soak the fruit in 75% alcohol for about 5 minutes, rinse it with sterile water for 3 to 4 times, and then soak it with 2.5% sodium hypochlorite for 3 minutes; plant the contact culture medium on the cutting part, and use the tissue imprinting method as a control to test whether the surface is disinfected. thorough;
[0164] b. Culture the culture medium in a constant temperature incubator at 28°C. Observe regularly, transfer the newly grown hyphae to the new solid medium in turn with an inoculation needle, purify each strain 2 to 3 times, and then transfer to the slant of the test tube to store two of each;
[0165] c. Preparation of bacterial suspension: Take a few slants of dominant strains cultured for one week, add ...
Embodiment 2
[0201] Strain preparation and industrial fermentation method:
[0202] i. The formula of liquid shake flask culture medium is PDA, pH 7. The initial pH value is 5.0. A total of 250ml was prepared, divided into 250mL Erlenmeyer flasks, 30ml per bottle, sterilized at 121°C for 30 minutes, then cooled to room temperature for use;
[0203] ii. Pick and transfer the cultured strains on the slant of the test tube into a 250ml Erlenmeyer flask equipped with a shaker culture medium, and place it in a shaking table at 28°C and 120 rpm for 24 hours;
[0204] iii. Prepare the seed culture medium according to the formula standard, pump the seed culture medium into the No. 2 500L first-level seed culture tank, and sterilize it by steam sterilization in real tank, the sterilization temperature is ≥121℃, and the sterilization time is ≥30min;
[0205] iv. After the sterilization of the first-level seed culture tank is completed, the temperature of the culture medium in the tank is lowered t...
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