New strain and method of producing pacilitaxel using said strain
一种紫杉醇、变种的技术,应用在使用真菌的方法、基于微生物的方法、生物化学设备和方法等方向,能够解决工业化生产困难、自然资源匮乏、耗费土地和人力等问题,达到满足临床用药的需要、保护自然资源、解决工艺问题的效果
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Embodiment 1
[0039] Example 1 Isolation of taxol-producing microorganisms from yew tree samples
[0040] We collected 310 samples from the branches, bark, leaves and root bark of seven Yunnan yew (Taxus yunnanensis) trees more than 300 years old in the mixed coniferous and broad-leaved forest at an altitude of 2500-3000 meters in Laojun Mountain, Lijiang area, Yunnan Province. Each was cut into small pieces of 1cm×1cm size, sterilized with 50-90% alcohol for 5 minutes, homogenized with a homogenizer, and then spread on PDA medium, cultured at 25°C for 4-5 days, and the grown The hyphae were transferred to the slope, cultured at 20°C-35°C for 4 days, and 1cm 2 Bacteria were inoculated in the fermentation medium (250ml Erlenmeyer flask capacity 100ml, medium formula: glucose 2%, soybean cake powder 0.5%, peanut cake powder 0.2%, MgSO 4 : 0.01%, KH 2 PO 4 : 0.05%), 25 ℃, 300rpm in the shaker culture 16 days, filter with double-layer gauze, get wet thallus, wet thallus is dried in oven (60 ...
Embodiment 2
[0041] Example 2 Purification and identification of new strains producing paclitaxel
[0042] Several paclitaxel-producing strains obtained in Example 1 were separated and purified on PDA medium plates for three generations (20-35°C, 4-10 days) to obtain the ST-026 strain. Strain ST-026 grows rapidly on PDA medium and produces abundant spores, which are usually black or olive green. Conidiophores grow directly from the agar medium, and branch conidia chains grow on the side shoots of the hyphae. The spore chains consist of 50-70 spores, the initial spores are produced directly from the sporophore, or from 1-2 spores are produced on the soil surface, spores are rod-shaped, oval-elliptic, about 22-31(25)X 9-13(11)UM in size, with 4-5 transverse septa, 2-3 ratio septa or oblique diaphragm. The initially formed young spores are narrowly oval, with dense, fine verrucous protrusions, such dense protrusions that the internal diaphragm of the spores cannot be seen clearly under a 10...
Embodiment 3
[0044] Embodiment 3 seed culture
[0045]Inoculate the obtained CGMCC No.0899 freeze-dried bacteria of Example 2 into a 250ml Erlenmeyer flask with 100ml seed culture medium (see Table 1), at 20~30°C, 100-300rpm shaker culture for 24-28 hours, to be ready Inoculated strains.
[0046] Table 1: Seed Media
[0047]
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