Anti-tumor protein extract from red onion bacteria and its preparation method and application
An anti-tumor and shallot technology, applied in the biological field, achieves good anti-tumor effect and good safety effect
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Embodiment 1
[0034] Preparation method one of red onion bacteria Boletus bicolor anti-tumor protein D1 of the present invention
[0035] 1) Dry the fruiting body of the red onion fungus at 50°C, crush it with a high-speed pulverizer, add distilled water with a liquid-to-material ratio of 10, shake at room temperature, 150r / min for 12h; centrifuge at 9000r for 20min, take the supernatant, add ammonium sulfate to make it saturated The concentration was 80%, and stood for 12 hours; after centrifugation, the precipitate was redissolved with distilled water, and after dialysis and desalination, the crude protein of the fruiting body of the red onion was obtained through freeze-drying;
[0036] 2) Dissolve the crude red onion protein obtained in step 1) in 50mM PH7.8 Tris-HCl buffer, and load it on the AKTApure protein purification instrument HiTrap TM Q HP column, linear elution with 0-1M NaCl solution, OD280 tracking and monitoring of each tube of eluent, drawing a curve according to the absor...
Embodiment 2
[0039] Example 2 The preparation method of Boletus bicolor anti-tumor protein D1 of the present invention II
[0040] 1) Dry the fruiting body of the red onion fungus at 50°C, crush it with a high-speed pulverizer, add distilled water with a liquid-to-material ratio of 10, shake at room temperature, 150r / min for 12h; centrifuge at 9000r for 20min, take the supernatant, add ammonium sulfate to make it saturated The concentration was 80%, and stood for 12 hours; after centrifugation, the precipitate was redissolved with distilled water, and after dialysis and desalination, the crude protein of the fruiting body of the red onion was obtained through freeze-drying;
[0041] 2) Dissolve the crude red onion protein obtained in step 1) in 50mM PH7.8 Tris-HCl buffer, and load it on the AKTApure protein purification instrument HiTrap TM Q HP column, use 0.35M, 0.45M and 1M NaCl solution for stepwise elution, OD280 tracking and monitoring of each eluate, draw a curve according to the ab...
Embodiment 3
[0044] Example 3 The preparation method of Boletus bicolor anti-tumor protein D1 of the present invention three
[0045] 1) Dry the fruiting body of the red onion fungus at 50°C, crush it with a high-speed pulverizer, add distilled water with a liquid-to-material ratio of 10, shake at room temperature, 150r / min for 12h; centrifuge at 9000r for 20min, take the supernatant, add ammonium sulfate to make it saturated The concentration was 80%, and stood for 12 hours; after centrifugation, the precipitate was redissolved with distilled water, and after dialysis and desalination, the crude protein of the fruiting body of the red onion was obtained through freeze-drying;
[0046] 2) Dissolve the crude red onion protein obtained in step 1) in 50mM PH7.8 Tris-HCl buffer, and load it on the AKTApure protein purification instrument HiTrap TM Q HP column, linear elution with 0-1M NaCl solution, OD280 tracking and monitoring of each tube of eluent, drawing a curve according to the absorban...
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