Preparation method of citrinin compound dicitrinone D and its application in lymphoma
A compound, the technology of citrinin, applied in the field of preparation of citrinin compound dicitrinoneD
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Embodiment 1
[0021] Example 1 Fermentation production and separation and purification of the compound
[0022] 1 Fermentation production
[0023] Fermentation culture of producing bacteria: According to the conventional method of cultivating microorganisms, take Penicillium citrinum IBPT-5 (preserved in the China Type Culture Collection on December 25, 2013, address: Wuhan University, Wuhan, the deposit number is : CCTCC NO: M 2013713) an appropriate amount, inoculate on the PDA solid slant medium and culture in a 28°C incubator for 4 days.
[0024] Take an appropriate amount of Penicillium citrinum IBPT-5 cultured on the slope for 4 days and inoculate it into 400mL of culture solution [Composition of culture solution (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] 1000mL Erlenmeyer flask, 28 ℃ static culture for 30 days, obtain mycelium and fermentation broth.
[0025] 2 Obtaining extract
[00...
Embodiment 2
[0032] Example 2 In vitro anti-tumor activity test
[0033] 1 Experimental samples and experimental methods
[0034] Preparation of the test sample solution The test sample is the pure compound separated and refined in the above-mentioned implementation 1. Accurately weigh an appropriate amount of sample and prepare a solution of the required concentration with DMSO for activity measurement.
[0035] The subculture of cell line and cell adopts tumor cell line. For tumor cells, use RPMI 1640 medium containing 10% FBS, and pass 5% CO at 37℃. 2 The incubator for relay culture.
[0036] Cell proliferation inhibitory activity test method
[0037] WST-1 method Take logarithmic growth phase lymphoma cells and adjust the cell density to 5×10 per milliliter 4 Cells were seeded in a 96-well cell culture plate at 100 μL per well, and 5% CO was injected at 37℃ 2 Cultivate overnight in the incubator. Aspirate the supernatant, add 100 μL of medium containing the sample, and continue to incubate for...
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