Mixed-source terpene compounds as well as separation method and application thereof
A technology of compounds and mixed terpenes, applied in the field of microbial medicine, can solve the problems of no neuraminidase activity, research, no literature reports, etc.
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Embodiment 1
[0025] Embodiment 1. Fermentative production and separation and purification of compounds 1, 2 and 3 shown in formula I:
[0026] The marine fungus Aspergillus ustus used in the present invention is isolated from sea squirt tissue, purified and cultivated on rice solid medium. The formula of rice solid medium is: rice 70g, corn steep liquor 0.2g, peptone 0.3g, yeast powder 0.5g, monosodium glutamate 0.6g , seawater 100ml.
[0027] Strain Aspergillus ustus (size 2.5 cm x 2.5 cm) was fermented in batches in rice solid medium in a 1000ml Erlenmeyer flask, the fermentation product was stirred and crushed, soaked and extracted with ethyl acetate for 3-4 times, and the combined extract was concentrated to obtain a fermented crude Extract.
[0028] The crude extract was subjected to vacuum silica gel column (200–300 mesh) chromatography, and the gradient was 20:1 to 1:1 (v / v) petroleum ether-ethyl acetate and the gradient was 20:1 to 1:1 ( v / v) Dichloromethane-methanol was used as ...
Embodiment 2
[0038] Example 2. Neuraminidase inhibitor activity.
[0039] Neuraminidase inhibitor screening kit (including neuraminidase detection buffer, neuraminidase, neuraminidase fluorescent substrate, Milli-Q water) was used to test neuraminidase inhibitory activity. Neuraminidase is a special glycoprotein on the surface of influenza virus and an important target of influenza drugs, so the screening of its inhibitors has become a common method for screening potential anti-influenza drugs.
[0040] The operation steps are as follows: Accurately weigh the sample of the pure product compound shown in formula I prepared in the above example, and prepare solutions of the required concentration with dimethyl sulfoxide (DMSO), respectively, and the concentrations of the positive control Tamiflu and the sample are both 200 μM .
[0041] Preparation of sample detection: Add 70 μl neuraminidase detection buffer, 10 μl neuraminidase, the neuraminidase inhibitor sample or the positive control Tam...
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