Method of extracting anti-intestinal cancer active protein from nostoc commune

A technology of active protein and cabbage, which is applied to the preparation methods of peptides, chemical instruments and methods, anti-tumor drugs, etc., can solve problems such as reports on anti-tumor active proteins of cabbage, which have not been seen, and achieve the effect of low cost.

Inactive Publication Date: 2013-04-24
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] After searching the existing literature, there is no report on the anti-tumor activity protein of Dipis radix

Method used

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  • Method of extracting anti-intestinal cancer active protein from nostoc commune
  • Method of extracting anti-intestinal cancer active protein from nostoc commune

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation method of the anti-intestinal cancer active protein of Chinese cabbage, the steps are as follows:

[0022] (1) Dried the natural ground vegetables and crushed them into powder with a grinder.

[0023] (2) Take 25g of ground vegetable powder, add 400ml of PBS buffer, stir on a stirrer, soak overnight at 4°C for 12 hours, centrifuge at 11000rpm for 25min, take the supernatant, add 150ml of PBS buffer to the filter residue, and do the same as above , the two extracts are combined to obtain the crude extract of Dipodella protein.

[0024] (3) Slowly add 2 times the volume of acetone pre-cooled at -20°C to the crude extract of the Chinese cabbage protein obtained in (2), stir slowly, then place it at -20°C for 1 hour, and centrifuge at 11,000 rpm for 30 minutes , discard the supernatant, collect the precipitate, place the precipitate at -20°C for 30 minutes, completely volatilize the acetone, and freeze-dry the precipitate to obtain 48 mg of crude protein of...

Embodiment 2

[0028] DLD1, SW620, HL-7702 cells in the logarithmic growth phase were treated with 1×10 4 per well into a 96-well culture plate. 37°C with 5% CO 2 After incubating in the incubator for 24 hours, the active protein of Dipis praecox was added at concentrations of 0.08, 0.16, 0.24, and 0.32 mg / ml respectively, and five replicate wells were set up, and PBS buffer was used as a control. After incubation for 48 hours, discard the medium in the wells, wash twice with PBS, add new medium, add 20 l of MTT solution (5 mg / ml) to each well, continue incubation for 4 hours, and terminate the culture. Carefully aspirate and discard the culture supernatant in the wells, add 150l DMSO to each well, and shake on a shaker for 10min to fully dissolve the crystals. At a wavelength of 570 nm, the light absorbance of each well was measured on an enzyme-linked immunosorbent monitor.

[0029] The active protein of Dipis pratense obtained in the present invention is used for the above-mentioned in...

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Abstract

The invention provides a method of extracting anti-intestinal cancer active protein from nostoc commune. The method comprises the following steps: crushing the pure natural nostoc commune, adding PBS (phosphate buffer solution) at low temperature, extracting overnight, then adding pre-cooled acetone to perform the low-temperature treatment to obtain the nostoc commune crude protein extracting solution; adding the ammonium sulfate powder in different saturation in the crude protein extracting solution, fractionally precipitating the protein, wherein the ammonium sulfate precipitate with saturation of 10-60% contains the nostoc commune anti-intestinal cancer active protein; then performing the Q anion column purification on the crude protein, wherein a penetrating solution contains the effective active protein. When the protein is used for treating the colorectal cancer cell for 48 hours, the IC50 value is 35 micrograms, and the protein has no obvious toxic and side effect for the normal cell; and the protein can be used for effectively inhibiting the proliferation of the colorectal cancer cell.

Description

technical field [0001] The invention relates to the extraction of protein, in particular to a method for extracting anti-intestinal cancer active protein from the ground vegetable. Background technique [0002] Ground vegetables, also known as ground ears, lichens, ground fungus, ground soft children, sweet potato skins, etc., are a combination of fungi and algae. There is a high output in Shanxi. It generally grows in rainy and humid places, dark black, a bit like soaked black fungus. Dirt vegetables are rich in protein, multivitamins and minerals such as phosphorus, zinc and calcium. Scientists at the Weizmann Institute in Israel have found that a component contained in the ground vegetable can inhibit the activity of acetylcholinesterase in the human brain, thereby producing a curative effect on Alzheimer's disease. Ground vegetables also have good nutritional and health value: 1. Lowering fat and improving eyesight is a very good low-fat nutrition and health food. It...

Claims

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Application Information

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IPC IPC(8): C07K14/405A61P35/00C07K1/36C07K1/30C07K1/18
Inventor 李卓玉晋小婷宋莉赵君宇李砧李文玲郭松佳
Owner SHANXI UNIV
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