Antitumor application of penicillium enol A2 from penicillium citrinum
A technology of Penicillium citrinum and IBPT-5, which is applied in the direction of anti-tumor drugs, methods based on microorganisms, and medical preparations containing active ingredients, etc. It can solve problems such as drugs that have not yet been seen, and achieve significant anti-tumor activity.
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Embodiment 1
[0020] Fermentative production and separation and purification of the compound of embodiment 1
[0021] 1 Fermentation production
[0022] Fermentation culture of production bacteria: according to the conventional method of cultivating microorganisms, take Penicillium citrinum ( Penicillium citrinum ) IBPT-5 (preserved in China Center for Type Culture Collection on December 25, 2013, address: Wuhan University, Wuhan, deposit number: CCTCC NO: M 2013713) appropriate amount, inoculated on PDA solid slant medium at 28 Cultivate in an incubator for 4 days.
[0023] Get the Penicillium citrinum ( Penicillium citrinum ) appropriate amount of IBPT-5, 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 0.3, NaCl 30.0 constant volume] in a 1000mL Erlenmeyer flask, after 30 days of static culture at 28°C, the mycelia and fermentation broth were obtain...
Embodiment 2
[0032] Example 2 Test of anti-tumor activity in vitro
[0033] 1 Experimental samples and experimental methods
[0034] 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.
[0035] Cell lines and cell subculture Tumor cell lines were used, and tumor cells were subcultured in DMEM medium containing 10% FBS at 37°C in an incubator filled with 5% carbon dioxide.
[0036] Cell Proliferation Inhibitory Activity Test Method
[0037] Tetrazolium salt (MTT) method Take tumor cells in the logarithmic growth phase and adjust the cell density to 2×10 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 2 for 4 hours in an incubator. Add 2 microliters of sample solution...
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