Secalonic acid H derived from penicillium oxalicum and application of secalonic acid H for preparing medicine for preventing human colon cancer
A technology of Penicillium oxalicum and human colon cancer cells, applied in the field of medicine, to achieve the effect of inhibiting the proliferation activity of human colon cancer cells
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Embodiment 1
[0019] Fermentative production and separation and purification of the compound of embodiment 1
[0020] 1 Fermentation production
[0021] Fermentation culture of production bacteria: according to the conventional method of cultivating microorganisms, take Penicillium oxalicum ( Penicillium oxalicum ) IBPT-6 (preserved in China Center for Type Culture Collection on December 25, 2013, address: Wuhan University, Wuhan, deposit number is: CCTCC NO: M 2013714) appropriate amount, inoculated on PDA solid slant medium at 28 Cultivate in an incubator for 3 days.
[0022] Penicillium oxalicum ( Penicillium oxalicum ) Appropriate amount of IBPT-6, inoculated into 400mL culture medium [Culture medium composition (g / L): mannitol 20.0, yeast extract 3.0, maltose 20.0, monosodium glutamate 10.0, glucose 10.0, KH 2 PO 4 0.5, MgSO 4 ·7H 2 O 0.3, NaCl 15.0, constant volume] in a 1000mL Erlenmeyer flask, after 30 days of static culture at 28°C, the mycelia and fermentation broth were ob...
Embodiment 2
[0031] Example 2 Test of Anti-tumor Activity in Vitro
[0032] 1 Experimental samples and experimental methods
[0033] Preparation of the tested sample solution The test sample is the pure product of the compound separated and refined in the above-mentioned implementation 1. Accurately weigh an appropriate amount of sample, and prepare a solution with the required concentration with methanol for activity measurement.
[0034] The subculture of cell lines and cells adopts tumor cell lines, and the tumor cells are treated with DMEM medium containing 10% FBS at 37°C with 5% CO 2 Subculture in an incubator.
[0035] Cell Proliferation Inhibitory Activity Test Method
[0036] Tetrazolium salt (MTT) method Take tumor cells in the logarithmic growth phase and adjust the cell density to 1×10 per ml 5 Cells were inoculated into 96-well cell culture plates at 200 microliters per well, and filled with 5% CO at 37°C 2for 4 hours in an incubator. Add 2 microliters of sample solution...
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