Method for preparing verticillium bacteria A and application
A technology of Verticillin and Verticillium, which can be applied in the field of medicine and can solve the problem of high mortality
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Embodiment 1
[0023] Example 1: Separation and purification of anti-tumor active compounds from the Amanita chrysalis infected by the parasite Alternaria sp. and its structural identification
[0024] experimental method:
[0025] 1. Extraction and purification: the wet weight of Amanita amanita fungus (including the fungal parasite Alternaria spp.) is 1500g, vacuum freeze-dried and crushed to obtain 120g dry powder, put it in a brown glass bottle and add 600ml ethyl acetate to extract at room temperature, from time to time Stir, filter after 24 hours, extract the residue with 4×500ml ethyl acetate, combine the filtrates, concentrate and recover the solvent by rotary evaporation to obtain 4.55g dark brown viscous extract, take 1.38g of the extract solvent to dissolve and put it on C8 reaction Phase preparation column (20*250mm) for separation and purification, the mobile phase is 10%-100% acetonitrile or methanol gradient elution, the cells are collected to screen for active elution peaks, ...
Embodiment 2
[0055] Example 2: Test of Verticillin A Proliferation Inhibitory Activity on Human Tumor Cell Lines
[0056] experimental method:
[0057] 1. Human tumor cell lines and sources: A total of ten tumor lines were made, namely erythroleukemia K562, breast cancer BCAP37, breast cancer MCF-7, cervical cancer HeLa, liver cancer SMMC 7721, liver cancer HepG2, lung adenocarcinoma SPC-A1, prostate cancer Cancer DU145, colorectal cancer SW620, and lymphoma Jurkat were all from Cancer Institute of Zhejiang University.
[0058] 2. Verticillin A and paclitaxel (products of Sichuan Taiji Pharmaceutical Co., Ltd.) were dissolved in DMSO to make a 1mg / ml mother solution, placed in a 4°C refrigerator, and diluted with culture medium to obtain a final concentration of 0, 0.01, 0.1, 1, 10ug / ml for use, MTT (AMRESCO company product) with phosphate-buffered saline (Phosphate-buffered Saline referred to as PBS) to prepare 5mg / ml mother solution, put in 4 ℃ refrigerator for later use.
[0059] 3. C...
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