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Black fungus protein extract and anti-tumor application thereof
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A technology of protein extract and black fungus, applied in antineoplastic drugs, peptide/protein components, applications, etc.
Active Publication Date: 2015-03-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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At present, the research on black fungus polysaccharides is relatively detailed, and there are many related functional research reports, but the separation of protein extracts from black fungus and the research on anti-tumor activity in vitro are still blank
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Embodiment 1
[0031] Embodiment 1, the preparation of black fungus protein extract
[0032] 1. Prepare black fungus fruiting bodies
[0033] Inoculate black fungus (Auricularia auricula) Au888 CFCC 89530 slant strains into the primary medium for activation, and culture at a constant temperature of 25°C, and inoculate it into the secondary seed medium after the mycelium is full of the test tube, and keep the temperature at 25°C Cultivate in the culture room until the hyphae are overgrown, inoculate the secondary species into the cultivation bag with the cultivation medium at 25°C for fungal growth, and then scratch the fungus after the fungus is full of the cultivation bag and move it into the greenhouse for fruiting Keep the humidity above 90% and the temperature at 20-25°C, collect the fruiting bodies of the first tide, and obtain the fruiting bodies of black fungus (Auricularia auricula) Au888 CFCC 89530.
[0034] Wherein, the culture medium used in this experiment is as follows:
[003...
Embodiment 2
[0042] Example 2, black fungus protein extract inhibits tumor cell proliferation experiment
[0043] 1.1 Tested cell lines
[0044] Human liver cancer HepG2 cells (purchased from ATCC, USA), human breast cancer MCF-7 cells (purchased from ATCC, USA) and human lung adenocarcinoma A-549 cells (purchased from ATCC, USA).
[0045] 1.2 Experimental method
[0046] 1.2.1 Cell lines and cell culture
[0047] Human liver cancer HepG2 cells, human breast cancer MCF-7 cells and human lung adenocarcinoma A-549 cells were activated and passaged according to conventional culture methods. Among them, the subculture and activation medium of human liver cancer HepG2 cells is DMEM-high glucose + 1% double antibody + 10% FBS (in DMEM-high glucose (Hyclone, SH30022.01B) a mixture of penicillin and streptomycin (penicillin 10000U / mL, streptomycin 10000μg / mL) and fetal bovine serum (FBS, Hyclone, SV30087.02), so that the final volume percentage of the former two is 1%, and the volume percentage...
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Abstract
The invention discloses a black fungus protein extract and anti-tumor application thereof. The black fungus protein extract is prepared according to the method including the following steps of 1) extracting by water after crushing black fungus sporocarp, collecting a water-soluble matter to obtain a black fungus water-soluble extract; 2) carrying out ammonium sulfate precipitation of which the saturation level is 80% on the black fungus water-soluble extract, collecting sediments, and obtaining the black fungus protein extract after dialyzing the sediments by deionized water. IC50 values are 6 mug / mL, 6 mug / mL, and 6 mug / mL respectively after a human liver cancer HepG2 cell, a human breast cancer MCF-7 cell and a pulmonary adenocarcinoma A-549 cell are processed by the black fungus protein extract for 72 hours; the inhibition ratio is increased along with time extension and increase of the concentration; apoptosis morphological changes of the cells can be observed under a light microscope.
Description
technical field [0001] The invention relates to black fungus protein extract and its anti-tumor application. Background technique [0002] Black fungus (Auricularia auricula), also known as fungus, ear, fungus, etc., belongs to the order Auricularia, the family Auricularia, and the genus Auricularia. It is a kind of fungus with high nutritional value for both medicine and food. Studies have shown that black fungus contains chemical components such as polysaccharides, adenosine, melanin, phospholipids and multivitamins, and has biological activities such as lipid-lowering, antithrombotic, anti-radiation, anti-mutation, and hypoglycemic. Liu Kechun and others studied the chemical components in black fungus, and used various chromatography methods such as silica gel and gel to separate and purify the chemical components in black fungus to obtain uridine (uridine), α-D glucopyranose (α-D -glucopyranose), cerebroside B (cerebroside B), and the anti-platelet aggregation activity ...
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