Preparation method of antitumor wheat germ proteins

A technology of wheat germ and wheat germ meal, which is applied to the preparation of anti-tumor protein and the field of deep processing of wheat, can solve the problems of unclear and inefficient realization of value-added transformation of wheat germ resources, unclear active ingredients, etc. Easy to scale application, significant increase in inhibitory activity

Active Publication Date: 2015-03-11
JIANGNAN UNIV
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Problems solved by technology

It has been reported that the water-soluble extract of wheat germ can significantly inhibit the growth of S-180 sarcoma, increase the weight of immune organs in tumor-bearing mice, restore the activity of peritoneal macrophages in tumor-bearing mice, and improve the macrophages of mice. The phagocytosis of phagocytic cells, and the water extract has a significant inhibitory effect on the growth of MDA-MB231 cells, but it is not clear which components play a role
Due to the unavoidable solvent residues and unclear active ingredients in fat-soluble extracts, salt-soluble extracts and fermented extracts, it brings certain uncertainty to the development of anti-tumor health food or drugs using wheat germ as raw materials, and it cannot be efficiently Realize the value-added transformation of wheat germ resources

Method used

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  • Preparation method of antitumor wheat germ proteins
  • Preparation method of antitumor wheat germ proteins
  • Preparation method of antitumor wheat germ proteins

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Embodiment 1

[0053] Add 400mL of water into a 500mL Erlenmeyer flask containing 40g of defatted wheat germ powder, extract on an air bath shaker at a speed of 200r / min for 3h, then centrifuge at 10000r / min, 4°C for 20min to obtain a supernatant, and use 200mL corresponds to solvent extraction for 1h, centrifuge under the same conditions, combine the supernatant, add 63.6g of solid ammonium sulfate to 20% saturation, stir for 40min, let stand for 3h, then centrifuge at 10000r / min, 4°C, 20min, Collect the supernatant, continue to add 72g of solid ammonium sulfate until the saturation is 40%, stir for 40min, let it stand for 3h, then centrifuge at 10000r / min, 4°C for 20min, discard the supernatant to collect the precipitate, add a small amount of water to redissolve the precipitate, Then put it into a dialysis bag, dialyze in water at 4°C for 24 hours, concentrate and freeze-dry to prepare WGWSP. Take 220 mg of wheat germ water-soluble protein freeze-dried sample (WGWSP), dissolve it in 10 mL...

Embodiment 2

[0056] Add 2000mL water into a 2500mL Erlenmeyer flask containing 200g defatted wheat germ powder, extract on an air bath shaker at a speed of 200r / min for 3h, then centrifuge at 10000r / min, 4°C for 20min to obtain a supernatant, and use 1000mL corresponds to solvent extraction for 1h, centrifuge under the same conditions, combine the supernatant, add 318g of solid ammonium sulfate to 20% saturation, stir for 40 minutes, let stand for 3h, then centrifuge at 10000r / min, 4°C, 20min, Collect the supernatant, continue to add 360g of solid ammonium sulfate until the saturation is 40%, stir for another 40 minutes, let stand for 3h, then centrifuge at 10000r / min, 4°C for 20min, discard the supernatant to collect the precipitate, and add a small amount of water to the precipitate to reconstitute Then put it into a dialysis bag, dialyze in water at 4°C for 24 hours, concentrate and freeze-dry to prepare WGWSP. Take 180 mg of wheat germ water-soluble protein freeze-dried sample (WGWSP),...

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Abstract

A preparation method of antitumor wheat germ proteins comprises the following steps: 1, taking fresh wheat germ, carrying out low temperature degreasing and crushing treatment, and extracting with water as a solvent; 2, removing insoluble substances in a mixed extract liquid obtained in step 1, carrying out grading precipitation of the obtained supernatant with ammonium sulfate step by step, separating out the obtained precipitate, re-dissolving the precipitate in water for dialysis, concentrating, and lyophilizing to obtain water-soluble wheat germ proteins; and 3, dissolving the water-soluble wheat germ proteins in a buffer solution, carrying out ion exchange chromatography, and carrying out gel filtering chromatography to obtain the antitumor wheat germ proteins. The method which treats water as an extracting solution is natural and healthy, does not cause the residual of toxic substances in the obtained product, is benefit for the improvement of the physiological activities of the obtained product, and has the advantages of simplicity and low cost; and the obtained product has substantial proliferation inhibition activities on the growth of human breast cancer cell strains, and has small toxicities to normal cells.

Description

technical field [0001] The invention relates to a preparation method of anti-tumor protein, in particular to a preparation method of wheat germ anti-tumor protein, which belongs to the deep processing technology of wheat. Background technique [0002] Wheat germ is a by-product of flour processing. The production of wheat flour in my country is huge, so the output of wheat germ is also considerable. In addition to high-quality protein, wheat germ also contains vitamin E, magnesium, pantothenic acid, phosphorus, thiamine, etc. It is a rare natural healthy food resource. [0003] Many kinds of anti-tumor active proteins have been found in plants. Foreign researchers have isolated polypeptide components from wheat germ fermentation extracts and proved that they have anti-tumor effects. The physiological activities of wheat germ protein and polypeptide, such as anti-oxidation, anti-aging, and anti-fatigue, have attracted extensive attention from scholars at home and abroad, but...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K1/36C07K1/34C07K1/30C07K1/18C07K1/16
Inventor 朱科学周会会郭晓娜周惠明彭伟
Owner JIANGNAN UNIV
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