Indole terpene speradine B derived from aspergillus oryzae and application
A kind of Aspergillus oryzae, application technology, applied in indoleterpene speradine B and its application field, can solve the problems such as no drugs have been seen, and achieve the effect of significant anti-tumor activity
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Embodiment 1
[0018] Fermentative production and separation and purification of the compound of embodiment 1
[0019] 1 Fermentation production
[0020] Fermentation culture of production bacteria: according to the conventional method of cultivating microorganisms, get Aspergillus oryzae ( Aspergillus oryzae ) IBPT-1 (preserved in China Center for Type Culture Collection on December 1, 2011, address: Wuhan University, Wuhan, deposit number is: CCTCC NO: M 2011437). Appropriate amount, inoculated on PDA solid slant medium and cultivated in 28°C incubator for 4 days.
[0021] Aspergillus oryzae ( Aspergillus oryzae ) appropriate amount of IBPT-1, 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, constant volume of seawater] in a 1000mL Erlenmeyer flask, after static culture at 28°C for 30 days, the mycelia and fermentation broth were obtained.
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Embodiment 2
[0029] Example 2 Test of anti-tumor activity in vitro
[0030] 1 Experimental samples and experimental methods
[0031] 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.
[0032] Cell lines and subculture of cells 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.
[0033] Cell Proliferation Inhibitory Activity Test Method
[0034] Lissamine rhodamine B (SRB) method Take Hela cells in the logarithmic growth phase and prepare them with fresh RPMI-1640 medium to a density of 2×10 per ml 5The cell suspension of each cell was inoculated in a 96-well cell culture plate at 200 microliters per well, and 2 microliters of different concen...
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